Clinicopathological Features of Intraductal Papillary Mucinous Neoplasms of Pancreas in a Tertiary Care Center: A 14 Year Retrospective Study.
Pancreatic surgeries are usually performed for inflammatory conditions and neoplasms. Intraductal Papillary Mucinous Neoplasm (IPMN) account for approximately 5-7% of all pancreatic neoplasms in western literature. To the best of our knowledge, there has been no published literature in Indian subcontinent on IPMN. To assess the spectrum of different pancreatic pathologies on pancreatic resection specimens. Also, to review and share the experience on the clinicopathological features of IPMNs in our institute. This was a 14 year retrospective study of all cases where pancreatic surgeries were done for pancreatic pathology. The slides and blocks of diagnosed cases of IPMNs were retrieved from the department archives, reviewed and a detailed study on the histopathological features was done. Among the 377 pancreatic surgical specimens, pancreatitis was the most common diagnosis followed by exocrine neoplasms and endocrine neoplasms. IPMN constituted 3.2 % of all pancreatic neoplasms. Histologically, the most common type was the gastric foveolar type. Pancreatobiliary type was aggressive and associated with an invasive component and had evidence of metastasis on follow up. IPMNs are rare neoplasms of pancreas with a male predominance. They are usually indolent except for the pancreatobiliary type which may have an aggressive course, often associated with an invasive adenocarcinoma component. Diligent follow up is recommended.


Synthesis of nucleoside boranophosphoramidate prodrugs conjugated with amino acids.
[structure: see text] Nucleoside boranophosphates and nucleoside amino acid phosphoramidates have been shown to be potent antiviral and anticancer agents with the potential to act as nucleoside prodrugs. A combination of these two types of compounds results in a boranophosphoramidate linkage between the nucleoside and amino acid. This new class of potential prodrugs is expected to possess advantages conferred by both types of parent compounds. Two approaches, specifically the H-phosphonate and oxathiaphospholane approaches, are described here to synthesize nucleoside boranophosphoramidate prodrugs conjugated with amino acids. The H-phosphonate approach involves a key intermediate, silylated nucleoside amino acid phosphoramidite 6, prepared from a series of reactions starting from nucleoside H-phosphonate in the presence of condensing reagent DPCP. Due to the lengthy procedure and the difficulties in removing DPCP from the final products, we switched to the oxathiaphospholane approach in which the DBU-assisted oxathiaphospholane ring-opening process constituted a key step for the generation of nucleoside amino acid boranophosphoramidates 24. We demonstrate that this key step did not cause any measurable C-racemization of boranophosphorylated amino acids 22. Diastereomers of compounds 24a-f were separated by RP-HPLC. An "adjacent"-type mechanism is proposed to explain the diastereomer ratio in the final products obtained via the oxathiaphospholane approach. A tentative assignment of configuration for the diastereomers was carried out based on the mechanism, molecular modeling, and (1)H NMR. Conclusively, the oxathiaphospholane methodology proved to be more facile and efficient than H-phosphonate chemistry in the preparation of the nucleoside amino acid boranophosphoramidate analogues that are promising as a new type of antiviral prodrugs.


Asociality and engagement in adult offenders in substance abuse treatment.
In therapeutic community models for drug treatment, individual recovery is principally structured around group meetings and positive peer influences. Research shows that asocial group members with resistant behaviors and attitudes are at risk for poor treatment outcomes and have the potential to adversely impact the therapeutic group. To gain a better understanding of the asocial client's role in the larger treatment process, in-prison treatment data were used: (1) to model and confirm a two-factor solution of asociality consistent with the literature, and (2) to examine the relationship between asocial clients and treatment engagement for validation of the two-factor model. Exploratory analysis resulted in a three-factor solution representing behavioral responsivity, cognitive distortion, and social disassociation dimensions. Nested ANOVA (i.e., clients nested within prison programs) demonstrated that asocial levels (low, medium, and high risk) predicted treatment engagement. Furthermore, comparisons among asocial risk levels indicated that high asocial clients reported significantly lower engagement levels when compared with low and medium asocial clients.


Absence of specific luteinizing hormone releasing hormone receptors in ovine, bovine and porcine ovaries.
Crude and membrane-enriched homogenates of unfrozen follicular and luteal tissue from cows, ewes and sows were assayed for the presence of specific luteinizing hormone releasing hormone (LHRH) receptors by one-point saturation analysis using [D-Ser-(TBU)6, des-Gly-NH2(10)] LHRH-EA as the labeled and unlabeled ligand. Pituitaries from cows, ewes, sows and rats, and rat ovaries served as positive controls and were assayed with each ovarian tissue assay. Scatchard analysis was used to determine binding affinity of pools of ovarian and pituitary tissue. Specific high-affinity LHRH receptors were found in the pituitaries of cows, ewes, sows and rats and in the rat ovary. In contrast, no specific LHRH binding was detected in follicular or luteal tissue of cows, ewes or sows. Thus, unlike the rat ovary which contains LHRH receptors, ovaries from these domestic species lack specific LHRH receptors.


In situ and non-invasive detection of specific bacterial species in oral biofilms using fluorescently labeled monoclonal antibodies.
Noninvasive in situ detection of suspected cariogenic bacterial species within dental biofilms could facilitate monitoring of the dynamic change of oral microbial flora and assist in the assessment of the treatment efficacy of therapeutic agents. In this study, we explore the possibility to use three well-characterized monoclonal antibodies (MAbs) against Streptococcus mutans, Actinomyces naeslundii, and Lactobacillus casei to identify these three important members of the oral microbial community in the complex environment of oral biofilms. These MAbs, which were conjugated to different fluorescent labels and visualized with confocal laser scanning microscopy (CLSM), proved to be an useful tool to identify the three species of interest (S. mutans, A. naeslundii, and L. casei) under various experimental conditions including in vitro and in vivo derived oral biofilms. Manifold addition of the MAbs on consecutive days did not alter the biofilm structure thus allowing monitoring of the same biofilm over extended time periods. Using this MAb-based method the effect of sucrose challenge on the biofilm composition and the distribution of S. mutans, A. naeslundii, and L. casei were examined. S. mutans was found to be the predominant species under the various biofilm conditions tested. These studies indicate that MAbs based bacterial detection with CLSM is a versatile tool which permits new insights into the ecology of oral biofilm development.


Effectiveness of a clinical pathway for complete denture adjustment.
This study was performed to evaluate the usefulness of a developed clinical pathway (CP) to determine the sequence of dental treatment in patients who are edentulous and use maxillary and mandibular complete dentures. Sixty-two edentulous patients with maxillary and mandibular complete dentures were randomly allocated either to with (31 subjects) or without (31 subjects) a CP. The main categories of the CP were the examination, instructions, practice, idle time, evaluation sheet, preparation, and other steps. The subcategories were made with respect to the main categories. The participants were categorized as dentists, assistants, or patients. There was no significant difference in mean chair time between with a CP (27.5 min) and without a CP (28.0 min). The CP led to a significant reduction in idle time (waiting in the chair or no treatment) of dentists, assistants, and patients. In contrast, the instances of medical examination, patients' instructions, and the number of patients reporting improvement of symptoms significantly increased. These results indicated that there is an opportunity to improve the complete denture treatment with a CP.


A Genome-Wide CRISPR/Cas9 Screen Reveals the Requirement of Host Sphingomyelin Synthase 1 for Infection with Pseudorabies Virus Mutant gD-Pass.
Herpesviruses are large DNA viruses, which encode up to 300 different proteins including enzymes enabling efficient replication. Nevertheless, they depend on a multitude of host cell proteins for successful propagation. To uncover cellular host factors important for replication of pseudorabies virus (PrV), an alphaherpesvirus of swine, we performed an unbiased genome-wide CRISPR/Cas9 forward screen. To this end, a porcine CRISPR-knockout sgRNA library (SsCRISPRko.v1) targeting 20,598 genes was generated and used to transduce porcine kidney cells. Cells were then infected with either wildtype PrV (PrV-Ka) or a PrV mutant (PrV-gD-Pass) lacking the receptor-binding protein gD, which regained infectivity after serial passaging in cell culture. While no cells survived infection with PrV-Ka, resistant cell colonies were observed after infection with PrV-gD-Pass. In these cells, sphingomyelin synthase 1 (SMS1) was identified as the top hit candidate. Infection efficiency was reduced by up to 90% for PrV-gD-Pass in rabbit RK13-sgms1KO cells compared to wildtype cells accompanied by lower viral progeny titers. Exogenous expression of SMS1 partly reverted the entry defect of PrV-gD-Pass. In contrast, infectivity of PrV-Ka was reduced by 50% on the knockout cells, which could not be restored by exogenous expression of SMS1. These data suggest that SMS1 plays a pivotal role for PrV infection, when the gD-mediated entry pathway is blocked.


Invasive strategy following fibrinolysis in ST-elevation acute myocardial infarction.
A recognized drawback of ST-elevation acute myocardial infarction (STEMI) after fibrinolysis is persistent coronary occlusion or a less than TIMI 3 flow. The present study describes the results of systematic pre-discharge coronary angiography and revascularization, whenever indicated, following fibrinolytic therapy for STEMI. Consecutive patients admitted with the diagnosis of STEMI between April 1, 2000 and April 30, 2002 were included in the study. Patients with contraindications to thrombolytic therapy and/or patients not eligible for angiography were excluded. All patients received "accelerated" treatment with alteplase and had a coronary angiography at least 24 hours later, in order to perform, if anatomically feasible, angioplasty with stenting. Angioplasty of non-infarct-related coronary arteries was allowed. The mortality, reinfarction and new revascularization rates were evaluated during index hospitalization and up to 30 days and 6 months. Eighty patients underwent cardiac catheterization at a median of 6.5 days following admission; in 86.3% of cases a patent infarct-related artery was found; in 71% of patients a coronary angioplasty was performed, with stenting in 88% of cases. Procedure-related complications were infrequent. No deaths occurred during hospitalization and at 30 days; at 6 months the mortality rate was 1.3%. In-hospital reinfarction occurred in 3.8% of patients, in 4% at 30 days and in 5.3% at 6 months. The rate of any new revascularization was 2.6% at 30 days and 11% at 6 months. Although obtained in a small observational study, our data, unlike those from previous studies, suggest that an invasive strategy after fibrinolysis in STEMI is safe and associated with low mortality and morbidity rates in the short and medium-terms.


Surface-exposed C-terminal amino acids of the small coat protein of Cowpea mosaic virus are required for suppression of silencing.
The small (S) coat protein of Cowpea mosaic virus (CPMV) has been identified previously as a virus-encoded suppressor of post-transcriptional gene silencing (PTGS). Deletions within the C-terminal 24 aa of this protein affect the yield and systemic spread of the virus, suggesting that the C-terminal amino acids of the S protein, which are exposed on the surface of assembled virus particles, may be responsible for the suppressor activity. To investigate this, versions of CPMV RNA-2 with deletions at the C terminus of the S protein were tested for their ability to counteract PTGS in leaf-patch tests. The results showed that the C-terminal 16 aa of the S protein are particularly important for suppressing PTGS and that these amino acids are virus-specific and cannot be substituted by the equivalent sequence from the related virus Bean pod mottle virus.


[Incarcerated diaphragmatic hernia presenting as colonic obstruction].
Diaphragmatic hernias in adults that are not categorized as hiatal hernias are rare. Hernias of the diaphragm itself appear in the perinatal age group and are defined as congenital hernias, presenting with severe respiratory distress and requiring immediate repair. There are a few asymptomatic post-partum cases in young children or adults. We present 3 cases of diaphragmatic hernia in men, aged 19, 20 and 74, respectively. 2 were admitted because of colonic incarceration with strangulation in the hernial sac. In the other case there was an incidental finding of incarcerated colon in a diaphragmatic hernia discovered in a patient who initially presented with unrelated thoraco-abdominal trauma. Contrary to the dominant clinical presentation as infant-respiratory distress, the predominant findings in adults are gastrointestinal complications. We believe successful treatment is more likely when there is a high level of awareness of the possibility of this kind of hernia in the adult, which can be a recurrence after perinatal hernia repair. It is essential to recognize that this is a combined thoraco-abdominal problem since it is often necessary to perform simultaneous laparotomy, thoracotomy and stoma formation.


p62/sequestosome 1 in human colorectal carcinoma as a potent prognostic predictor associated with cell proliferation.
p62/sequestosome 1 (p62) is a multi-domain protein that functions as a receptor for ubiquitinated targets in the selective autophagy and serves as a scaffold in various signaling cascades. p62 have been reported to be up-regulated in several human malignancies, but the biological roles and significance of p62 are still poorly understood in colorectal carcinoma. We immunohistochemically evaluated p62 in 118 colorectal adenocarcinoma and 28 colorectal adenoma cases. We used four colon carcinoma cells (HCT8, HT29, COLO320, and SW480) in the in vitro studies. p62 immunoreactivity was detected in 11% of colorectal adenoma cases and 31% of adenocarcinoma cases, while it was negligible in the normal epithelium. The immunohistochemical p62 status was significantly associated with synchronous liver metastasis, and it turned out to be an independent adverse prognostic factor in colorectal cancer patients. Following in vitro studies revealed that HCT8 and HT29 cells transfected with p62-specific siRNA showed significantly decreased cell proliferation activity, whereas COLO320 and SW480 cells transfected with p62 expression plasmid showed significantly increased cell proliferation activity. The p62-mediated cell proliferation was not associated with the autophagy activity. These findings suggest that p62 promotes the cell proliferation mainly as a scaffold protein, and that the p62 status is a potent prognostic factor in colorectal carcinoma patients.


BASILAR ARTERY DISEASE-Clinical Manifestations.
Patients with basilar artery disease show their first symptoms by characteristic ischemic attacks. If the attending physician correctly diagnoses the condition and initiates anticoagulant therapy before a catastrophic episode occurs, there is reason to believe that the disease may be arrested temporarily.


Temperature preconditioning is optimal at 26° C and confers additional protection to hypothermic cardioplegic ischemic arrest.
We have recently shown that brief episodes of hypothermic perfusion interspersed with periods of normothermic perfusion, referred to as temperature preconditioning (TP), are cardioprotective and can be mimicked by consecutive isoproterenol/adenosine treatment. Here we investigate the optimal temperature for TP and whether TP further enhances protection provided by hypothermic ischemia with or without polarized cardioplegic arrest. Three experimental groups of Langendorff-perfused rat hearts were used. In the first group, hearts were subjected to three episodes of hypothermic perfusion at 7, 17, 26 and 32°C during the TP protocol, followed by 30 min normothermic index ischemia and 60 min reperfusion (37°C). Protein kinase A (PKA) activity and cyclic AMP (cAMP) concentrations were measured prior to index ischemia. In the second group, TP (26°C) hearts were subjected to two hours hypothermic index ischemia at 26°C and two hours normothermic reperfusion. In the third group, TP (26°C) hearts or hearts treated with isoproterenol/adenosine (pharmacological simulation of TP) were subjected to four hours hypothermic index ischemia with procaine-induced polarized cardioplegia at 26°C followed by two hours normothermic reperfusion. Hemodynamic function recovery, lactate dehydrogenase release and infarct size were used to assess cardioprotection. TP at 26°C resulted in highest cardioprotection, increased cAMP concentration and PKA activity, while TP at 7°C exacerbated ischemia/reperfusion damage, and had no effect on cAMP concentration or PKA activity. TP at 26°C also protected hearts during hypothermic ischemia with or without polarized cardioplegia. Isoproterenol/adenosine treatment conferred additional protection similar to TP. In conclusion, the study shows that TP-induced cardioprotection is temperature dependent and is optimal at 26°C; TP confers additional protection to hypothermia and polarized cardioplegia; and that the pharmacological treatment based on the mechanism of TP (consecutive isoproterenol/adenosine treatment) is a potential cardioprotective strategy that can be used during heart surgery and transplantation.


Age and indications to SLIT.
Clinical efficacy of sublingual immunotherapy (SLIT) has been investigated during the last 20 years and results of several meta-analyses are available, showing clinical efficacy of SLIT in children both in allergic asthma and in rhinitis, but strict recommendations are not possible under current evidence. Minimum age for starting SLIT is not clearly defined but several position paper and guidelines indicate a lower limit of 5 years of age. Guidelines on allergic rhinitis suggests SLIT in patients not well controlled with drugs or those who refuse to use drugs. Additional effects are prevention of new sensitizations (evidence IIa) and prevention of asthma in patients with allergic rhinitis (evidence I b). Studies on efficacy of SLIT in asthmatic children are discordant, but the different relevance of allergic and non allergic triggers of symptoms could explain the discordant results obtained in studies on SLIT and asthma, particularly when pooling short and long term studies. Data on efficacy and safety of SLIT are accruing for atopic dermatitis, food allergy and latex allergy, but at the current state of knowledge, SLIT remains an approach reserved to research, and no recommendations can be established. Some studies demonstrate that SLIT is safe in children below 5 years of age, with a lower limit of 3 years.


Sternal fracture: isolated lesion versus polytrauma from associated extrasternal injuries--analysis of 1,867 cases.
Sternal fractures fall into two distinct categories as follows: (1) isolated sternal fracture (ISF) without associated injury and (2) polytrauma sternal fracture (PSF) with associated extrasternal injury. PSF can be sufficiently severe to require hospitalization, while ISF is usually a mild insult manageable in an ambulatory setting. Nonetheless, most patients with ISF are hospitalized. The disconnect between treatment based on existing evidence and actual clinical practice may be caused in large part by the small patient cohorts in published studies conducive to inaccurate conclusions. This article addresses the issue by analyzing prospectively collected data on a far larger population than hitherto available. The Israeli National Trauma Registry (INTR) collects data prospectively on patients admitted to trauma centers in Israel. We analyzed the INTR database on sternal fractures for demographics, mechanisms and severity of injury, diagnostic evaluation, treatment, and outcomes. Between 1997 and 2008, the INTR received data on 1,867 consecutive patients hospitalized with sternal fractures. The injury was sustained most often during motor vehicle accidents (84.1%), followed by falls (10.3%) and other blunt or penetrating mechanisms (5.6%). ISF was sustained in 26.4%, and PSF was sustained in 73.6%. Associated injuries involved most parts of the body. Cardiac contusions or lacerations were diagnosed in 1.8% of the 1,867 patients. ISF was associated with a lower incidence of cardiorespiratory compromise, more favorable trauma scores (Glasgow Coma Scale [GCS] score > 14, Revised Trauma Score > 11), and use of fewer intensive care facilities or operations compared with PSF. Endotracheal intubation, chest tube insertion, or thoracotomy was not performed in any ISF patient but was performed in 16.9% of the PSF cohort. The differences between ISF and PSF were statistically significant in all analyzed indices of injury severity. This study produced compelling evidence that ISF is an identifiable and mild injury. Consequently, pain, the major clinical manifestation of ISF, can usually be treated in outpatient settings. To avoid the inconvenience, risk of complications, and cost associated with hospitalization, discharge from the emergency department merits serious consideration. Prognostic and epidemiologic study, level II.


Actin remodeling driven by circLIMA1: sperm cell as an intriguing cellular model.
CircRNA cargo in spermatozoa (SPZ) participates in setting cell quality, in terms of morphology and motility. Cannabinoid receptor CB1 activity is correlated with a proper spermatogenesis and epididymal sperm maturation. Despite CB1 promotes endogenous skill to circularize mRNAs in SPZ, few notions are reported regarding the functional link between endocannabinoids and spermatic circRNA cargo. In CB1 knock-out male mice, we performed a complete dataset of spermatic circRNA content by microarray strategy. Differentially expressed (DE)-circRNAs, as a function of genotype, were identified. Within DE-circRNAs, we focused the attention on circLIMA1, as putative actin-cytoskeleton architecture regulator. The validation of circLIMA1 dependent-competitive endogenous RNA (ceRNA) network (ceRNET) in in vitro cell line confirmed its activity in the regulation of the cytoskeletal actin. Interestingly, a dynamic actin regulation in SPZ nuclei was found during their epididymal maturation. In this scenario, we showed for the first time an intriguing sperm nuclear actin remodeling, regulated via a ceRNET-independent pathway, consisting in the nuclear shuttling of circLIMA1-QKI interactome and downstream in Gelsolin regulation. In particular, the increased levels of circLIMA1 in CB1 knock-out SPZ, associated with an inefficient depolymerization of nuclear actin, specifically illustrate how endocannabinoids, by regulating circRNA cargo, may contribute to sperm morpho-cellular maturation.


Percutaneous Coronary Intervention versus Coronary Artery Bypass Grafting in Patients with Diabetic Nephropathy and Left Main Coronary Artery Disease.
Patients with diabetic nephropathy and unprotected left main (LM) coronary artery disease suffer from high cardiovascular morbidity and mortality. Although surgical revascularization is currently recommended in this special patient population, the optimal revascularization method for this distinct patient group has remained unclear. We collected 99 consecutive patients with unprotected LM disease and diabetic nephropathy, including 46 patients who had undergone percutaneous coronary intervention (PCI), and 53 who had coronary artery bypass grafting (CABG), with a mean age of 72 ± 10; with 80.8% male. Diabetic nephropathy was defined as overt proteinuria (proteinuria > 500 mg/day) and estimated glomerular filtration rate (eGFR) by the modified Modification of Diet in Renal Disease (MDRD) equation of less than 60 mL/min/1.73 m2. The baseline characteristics, angiographic results and long-term clinical outcomes were retrospectively analyzed. The baseline characteristic of all patients were similar except for smokers, low density lipoprotein (LDL) level and extension of coronary artery disease involvement. The median follow-up period was 3.8 years. There were 73 patients (74%) considered as high risk with additive European System for Cardiac Operative Risk Evaluation (EuroSCORE) ≥ 6. During follow-up period, the long term rate of all-cause death (PCI vs. CABG: 45.7% vs. 58.5%, p = 0.20) and all-cause death/myocardial infarction (MI)/stroke (PCI vs. CABG: 52.2% vs. 60.4%, p = 0.41) were comparable between the PCI and CABG group, whereas the repeat revascularization rate was significantly higher in the PCI group (PCI vs. CABG: 32.6% vs. 9.4%, p < 0.01). eGFR remained an independent predictor for all-cause death [hazard ratio: 0.97, 95% confidence interval: 0.96 to 0.99; p = 0.002] in multivariate logistic regression. In the real-world practice of high-risk patients with unprotected LM disease and diabetic nephropathy, we found that PCI was a comparable alternative to CABG in terms of long-term risks of all-cause death/MI/stroke, with significantly higher repeat revascularization rate. Given the small patient number and retrospective nature, our findings should be validated by larger-scale randomized studies.


Elucidating the dynamic immune responses within the ocular mucosa of rainbow trout (Oncorhynchus mykiss) after infection with Flavobacterium columnare.
The eye of vertebrates is constantly faced with numerous challenges from aquatic or airborne pathogens. As a crucial first line of defense, the ocular mucosa (OM) protects the visual organ from external threats in vertebrates such as birds and mammals. However, the understanding of ocular mucosal immunity in early vertebrates, such as teleost fish, remains limited, particularly concerning their resistance to bacterial infections. To gain insights into the pivotal role of the OM in antibacterial immunity among teleost fish, we developed a bacterial infection model using Flavobacterium columnare in rainbow trout (Oncorhynchus mykiss). Here the qPCR and immunofluorescence results showed that F. columnare could invade trout OM, suggesting that the OM could be a primary target and barrier for the bacteria. Moreover, immune-related genes (il-6, il-8, il-11, cxcl10, nod1, il1-b, igm, igt, etc.) were upregulated in the OM of trout following F. columnare infection, as confirmed by qPCR, which was further proved through RNA-seq. The results of transcriptome analyses showed that bacterial infection critically triggers a robust immune response, including innate, and adaptive immune-related signaling pathways such as Toll-like, NOD-like, and C-type lectin receptor signaling pathway and immune network for IgA production, which underscores the immune role of the OM in bacterial infection. Interestingly, a substantial reduction in the expression of genes associated with visual function was observed after infection, indicating that bacterial infection could impact ocular function. Overall, our findings have unveiled a robust mucosal immune response to bacterial infection in the teleost OM for the first time, providing valuable insights for future research into the mechanisms and functions of ocular mucosal immunity in early vertebrate species.


Prognostic Value of 18F-Fluorocholine PET Parameters in Metastatic Castrate-Resistant Prostate Cancer Patients Treated with Docetaxel.
The availability of new treatments for metastatic castrate-resistant prostate cancer (mCRPC) patients increases the need for reliable biomarkers to help clinicians to choose the better sequence strategy. The aim of the present retrospective and observational work is to investigate the prognostic value of 18F-fluorocholine (18F-FCH) positron emission tomography (PET) parameters in mCRPC. Between March 2013 and August 2016, 29 patients with mCRPC were included. They all received three-weekly docetaxel after androgen deprivation therapy, and they underwent 18F-FCH PET/computed tomography (CT) before and after the therapy. Semi-quantitative indices such as maximum standardized uptake value (SUVmax), mean standardized uptake value (SUVmean) with partial volume effect (PVC-SUV) correction, metabolically active tumour volume (MATV), and total lesion activity (TLA) with partial volume effect (PVC-TLA) correction were measured both in pre-treatment and post-treatment 18F-FCH PET/CT scans for each lesion. Whole-body indices were calculated as sum of values measured for each lesion (SSUVmax, SPVC-SUV, SMATV, and STLA). Progression-free survival (PFS) and overall survival (OS) were considered as clinical endpoints. Univariate and multivariate hazard ratios for whole-body 18F-FCH PET indices were performed, and p < 0.05 was considered as significant. Cox regression analysis showed a statistically significant correlation between PFS, SMATV, and STLA. No correlations between OS and 18F-FCH PET parameters were defined probably due to the small sample size. Semi-quantitative indices such as SMATV and STLA at baseline have a prognostic role in patients treated with docetaxel for mCRPC, suggesting a potential role of 18F-FCH PET/CT imaging in clinical decision-making.


Cooperativity of actin and microtubule elements during replication of respiratory syncytial virus.
Many paramyxoviruses appear to require cytoskeletal elements for particular steps in the virus life cycle. Measles virus and Sendai virus exhibit a requirement for microtubules in replication in vitro, whereas parainfluenza virus type 3 and RSV require actin for replication. To further elucidate the role of cytoskeletal function and rearrangement in the viral life cycle of RSV, we investigated the efficiency of virus entry, transcription, replication, and budding in the presence of a variety of pharmacological agents that stabilize or depolymerize actin or microtubules. We found that alteration of microtubule or actin function resulted in blocks at entry, formation of cell-associated virus, virus release, local cell-to-cell spread, and syncytium formation. Actin and microtubules act in cooperation to facilitate replication of RSV, although microtubules play a dominant role in the formation of cell-associated virus while actin plays a more prominent role in virus release.


Performances of linear tools and models for error detection and concealment in subband image transmission.
In this paper, we investigate the performances of Gaussian modeling and linear prediction tools for error detection and concealment in the transmission of still images. We consider the transmission of subband encoded images through two types of channels. We model the residual correlation between subband coefficients by considering them as jointly Gaussian variables. The first transmission medium considered is a packet-oriented channel, where some packets are lost during transmission. The problem is to estimate the values of missing coefficients. In this case, particular care must be taken while evaluating correlation matrices from incomplete data. The other system considered is based on a discrete memoryless noisy channel affecting the data being transmitted. The challenge is here first to determine the locations of the errors--which is done through hypotheses tests--and then to replace them by estimates based on their neighbors. The reconstruction via linear prediction is shown to give better results than median filtering based reconstruction. Error detection through this Gaussian model also shows promising results, in particular when channel statistics are taken into account in a joint source-channel decoding framework.


Vaccination of eels (Anguilla japonica and Anguilla anguilla) against Anguillicola crassus with irradiated L3.
The original host of the swimbladder nematode Anguillicola crassus, the Japanese eel (Anguilla japonica) and the recently colonized European eel (Anguilla anguilla) were immunized with 40 irradiated (500 Gy) 3rd-stage larvae (L3) of this parasite and challenged with an infection of 40 normal L3. The immunization induced a significant reduction of the number of adult worms developing from the challenge infection in A. japonica, but not in A. anguilla. The induced resistance (calculated using the relation of the number of adult worms in immunized eels and in non-immunized control eels) in A. japonica was 87.3%+/-30.4%. Following a single infection, the percentage of adult worms found in A. japonica was lower as compared to A. anguilla, and the few adult worms were much smaller, revealing a lower susceptibility of A. japonica to A. crassus in comparison to A. anguilla. Both eel species developed an antibody response against A. crassus, but the level of antibody responses was not positively correlated with the protection against infection, suggesting that the antibody response is not a key element in resistance of eels against A. crassus. This study suggests that the original host of A. crassus is able to mount efficient protective immune responses against its parasite, whereas the newly acquired host seems to lack this ability.


Cardiac remodelling and myocardial recovery: lost in translation?
The insight that decreases in left ventricular (LV) volume and mass occur secondary to the recovery of the myocardium at the cellular and molecular level has engendered a wider appreciation of the importance of LV remodelling as a mechanism for worsening heart failure. Despite these recent insights into the recognition of the importance of LV reverse remodelling in heart failure, many clinicians do not consider simple measurements of LV structure (i.e. LV volume) in their routine clinical decision-making process, preferring instead to rely on measurements of LV function [e.g. ejection fraction (EF)] when making decisions about medical and surgical treatment options. Although there are probably multiple reasons of why the use of LV volumes has not gained wider acceptance in day-to-day clinical management of heart failure patients, the most likely reason is that clinicians remain extremely comfortable using the LVEF to assess their heart failure patients. Importantly, LV volumes predict outcome more reliably than does the EF. Moreover, knowledge regarding LV volumes is extremely useful in optimizing patient selection for surgical and device therapies. Based on the foregoing arguments, we suggest that it is time to begin developing individualized clinical strategies based upon a consideration of the important role that LV remodelling plays in the pathogenesis of heart failure, and that we begin to incorporate measurements of LV volume and mass into the clinical decision-making process.


Therapy-related leukemia and myelodysplastic syndrome in breast cancer patients treated with cyclophosphamide or anthracyclines.
Accumulation of data regarding therapy-related leukemia (TRL) or myelodysplastic syndrome (t-MDS) is critical for assessing the risk of developing such diseases and for subsequent decision-making processes for better treatment. We evaluated the clinical characteristics of patients with TRL/t-MDS diagnosed at the National Cancer Center Hospital between January 1989 and September 1997. This report is concerned with those patients who initially had been treated with chemotherapeutic agents for breast cancer. Thirteen patients (median age, 55 years) developed TRL (n = 4) or t-MDS (n = 9). The median interval between the development of TRL/t-MDS and initial treatment was 94 months (range 23-190 months). For the primary therapy, all patients had received intense and prolonged treatment with cyclophosphamide (CPA) and/or anthracyclines including doxorubicin (DOX), with a median cumulative dose of 55 g/body (range 16.4-288.5 g) for CPA and 480 mg/m2 (range 395-625.5 mg/m2) for DOX. Seven patients were subsequently treated by chemotherapy and one received an allogeneic bone marrow transplantation. Clinicians must remain alert to the risks associated with unproven medical practices which include long-term administration of alkylating agents. Selected patients with TRL/t-MDS may respond to intense salvage combination chemotherapy.


Optical constant determination of an anisotropic thin film via polarization conversion.
This study presents a simple method for determining the optical constants of an anisotropic thin film. The sensitivity of enhanced polarization conversion reflectance to optical constants is also calculated and analyzed. Based on the sensitivity calculation, the principal indices and columnar tilt angle can be derived from the polarization conversion reflectance angular spectrum.


Investigation of drug metabolism in various segments of small intestine in the rat.
In the extrahepatic drug metabolism the intestinal tract can play an important role. These experiments were designed to study the biotransformation of p-nitrophenol (PNP) in the small intestine in the rat. Various segments of the small intestine (proximal and distal jejunum, terminal ileum) were perfused with isotonic solution in vivo containing different concentrations of PNP (20-100-500-1000 μM) and the concentrations of metabolites (PNP-G: p-nitrophenol glucuronide, PNP-S: p-nitrophenol sulfate) were determined in the perfusion medium. It was found a decreasing tendency in the glucuronidation from the proximal to distal segment of the small intestine: e.g. 430 nmol, 240 nmol, and 100 nmol PNP-G appeared in the perfusion medium in the proximal, distal jejunum and in the terminal ileum, respectively, when 500 μM PNP was luminally perfused for 90 minutes. Similar ratio was found at the luminal perfusion of other PNP-concentrations, too. Luminal appearance of sulfoconjugate of PNP was considerably lower and no clear gradient tendency in the formation of PNP-S could be detected in the small intestine from the proximal to distal segment. Our results show that there are considerable differences in drug metabolism in various segments of the small intestine. We have found a gradient conjugating activity from proximal to distal segment of small intestine in the glucuronidation of PNP.


Impartial institutions, pathogen stress and the expanding social network.
Anthropologists have documented substantial cross-society variation in people's willingness to treat strangers with impartial, universal norms versus favoring members of their local community. Researchers have proposed several adaptive accounts for these differences. One variant of the pathogen stress hypothesis predicts that people will be more likely to favor local in-group members when they are under greater infectious disease threat. The material security hypothesis instead proposes that institutions that permit people to meet their basic needs through impartial interactions with strangers reinforce a tendency toward impartiality, whereas people lacking such institutions must rely on local community members to meet their basic needs. Some studies have examined these hypotheses using self-reported preferences, but not with behavioral measures. We conducted behavioral experiments in eight diverse societies that measure individuals' willingness to favor in-group members by ignoring an impartial rule. Consistent with the material security hypothesis, members of societies enjoying better-quality government services and food security show a stronger preference for following an impartial rule over investing in their local in-group. Our data show no support for the pathogen stress hypothesis as applied to favoring in-groups and instead suggest that favoring in-group members more closely reflects a general adaptive fit with social institutions that have arisen in each society.


Pressure and shear differentially alter human articular chondrocyte metabolism: a review.
Homeostasis of articular cartilage depends in part on mechanical loads generated during daily activity whereas inappropriate joint loads result in focal degeneration of cartilage, as occurs in osteoarthritis. We will review results of a series of questions regarding the effects of two types of mechanical loads-intermittent hydrostatic pressure and shear stress-on adult human articular chondrocytes in high-density monolayer culture. Intermittent hydrostatic pressure increased aggrecan and Type II collagen gene expression in normal chondrocytes and induced changes in the cell-associated proteins of normal and osteoarthritic chondrocytes. Hydrostatic pressure also counteracted inhibitory effects of bacterial lipopolysaccharide on matrix protein expression by cultured chondrocytes. Application of shear stress to osteoarthritic chondrocytes increased the release of the proinflammatory mediator, nitric oxide, decreased aggrecan and Type II collagen expression, and induced molecular changes associated with apoptosis whereas hydrostatic pressure increased matrix macromolecule expression. The findings show that the types of load comprising the mechanical loading environment of articular cartilage considerably alter chondrocyte metabolism and suggest that mechanical stimulation may be used for in vitro or in vivo approaches for cartilage engineering.


[A new study method: the perineal scan in obstetrics and gynecology].
For gynecological/obstetrical ultrasound examination the abdominal wall constituted the first and for a long time the only acoustical window that could be used to visualize the female reproductive organs. Only recently transvaginal/transrectal endosonography has opened new approaches in this diagnostic field. Apart from the above-mentioned procedures, the female perineum can be regarded as a useful window to demonstrate the cervix uteri, the bladder and the urethra. At the Universitäts-Frauenklinik Heidelberg this examination procedure was used since June, 1985, to evaluate its clinical reliability in obstetrics and gynecology.


An international study of trainee-trained transitions: Introducing the transition-to-trained-doctor (T3D) model.
Background: Throughout their careers, doctors and other healthcare professionals experience numerous transitions. When supporting transitions, opportunities for development and learning should be maximized, while stressors having negative impacts on well-being should be minimized. Building on our international data, this study aimed to develop a conceptual model of the trainee-trained transition (i.e. the significant transitions experienced by doctors as they complete postgraduate training moving from trainee/resident status to medical specialist roles).Methods: Employing Multiple and Multidimensional Transitions (MMT) theory and current conceptualizations of clinical context, this study undertook secondary analysis of 55 interviews with doctors from three countries (Netherlands, Cananda and the UK) undergoing trainee-trained transitions.Results: Through this analysis, the Transition-To-Trained-Doctor (T3D) conceptual model has been developed. This model takes into consideration the multiple contexts and multiple domains in which transitions take place.Discussion: This model is significant in that it has several uses and is applicable across countries: to remind doctors, managers and medical educators of the complexity of transitions; to frame and facilitate supportive conversations; and as a basis to teach about transitions.


AMP-activated protein kinase signaling protects oligodendrocytes that restore central nervous system functions in an experimental autoimmune encephalomyelitis model.
AMP-activated protein kinase (AMPK) signaling is reported to protect neurons under pathologic conditions; however, its effect on oligodendrocytes (OLs) remains to be elucidated. We investigated whether AMPK signaling protects OLs to restore central nervous system (CNS) functions in an experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. Increased inflammation and demyelination in the CNS and peripheral immune responses were consistent with the observed clinical impairments in EAE animals, which were attenuated by treatment with metformin compared with vehicle. In addition, expressions of neurotrophic factors and of signatory genes of OL lineages were increased in the CNS of metformin-treated EAE animals. Likewise, metformin attenuated inflammatory response and enhanced expressions of neurotrophic factors, thereby protecting OLs via AMPK activation in mixed glial cultures stimulated with lipopolysaccharide/interferon γ in vitro, as evidenced by analysis of the expression of signatory genes of O1(+)/MBP(+) OLs and their cellular populations. Metformin also attenuated oxidative stress and malondialdehyde-containing protein levels, with corresponding induction of antioxidative defenses in OLs exposed to cytokines via AMPK activation. These effects of metformin were evident in the CNS of EAE animals. These data provide evidence that AMPK signaling is crucial to protect OLs and, thus, CNS functions in EAE animals. We conclude that AMPK activators, including metformin, have the potential to limit neurologic deficits in multiple sclerosis and related neurodegenerative disorders.


Mechanisms Involved in the Stimulatory and Inhibitory Effects of 5-Hydroxytryptamine on Vagal Mechanosensitive Afferents in Rat Lung.
Mechanosensitive vagal afferents in the lung, rapidly and slowly adapting receptors (RARs and SARs, respectively), play an important role in eliciting the reflexes that regulate the normal airway function. A profound bronchoconstrictive effect of 5-hydroxytryptamine (5-HT) has been extensively reported in various animal species, but its influence on the SAR and RAR activity is not known. This study investigated the effect of 5-HT on these receptors, and the possible mechanisms involved. Single-fiber activities of these afferents were measured in anesthetized, open-chest, and mechanically ventilated rats. Our results showed that intravenous injection of 5-HT evoked a consistent and pronounced stimulation of phasic RARs. In contrast, 5-HT generated an inconsistent and paradoxical action on SARs: no effect in 29% (5 of 17) of the SARs; stimulation in 35% (6 of 17); and inhibition in the remainder. These responses of both RARs and SARs to 5-HT were reproducible and dose-dependent. After the injection of a high dose of 5-HT (16 μg/kg), the receptor responses slowly reached a peak (after ∼8 s) and returned toward the baseline in ∼20 s, accompanied by a consistent increase in total pulmonary resistance and a decrease in dynamic lung compliance in a temporal pattern very similar to the increased receptor activity. When these changes in lung mechanics induced by 5-HT were prevented by pretreatment with salbutamol, a β2 adrenergic receptor agonist, the delayed responses of both RARs and SARs to 5-HT were also abolished, except that the immediate stimulatory effect on a subset of RARs, the silent RARs, was not affected. In conclusion, 5-HT generated a delayed stimulatory effect on RARs and a paradoxical effect on SARs, which resulted primarily from the 5-HT-induced changes in mechanical properties of the lung.


Histology-guided high-resolution AP-SMALDI mass spectrometry imaging of wheat-Fusarium graminearum interaction at the root-shoot junction.
Fungal pathogens like Fusarium graminearum can cause severe yield losses and mycotoxin contamination of food and feed worldwide. We recently showed its ability to systemically colonize wheat via root infection. However, the molecular response of wheat to Fusarium root rot (FRR) infection and systemic spread is still unknown. As a molecular camera, mass spectrometry (MS) imaging combines label-free and multiplex metabolite profiling with histopathology. Atmospheric-pressure (AP)-SMALDI-MS imaging was combined with optical microscopy to study wheat-F. graminearum interaction at the root-shoot junction, which is a crucial line of defense against a pathogen that can invade all distal plant parts. To scope the functional, temporal and local aspects of FRR disease spread, metabolic changes were simultaneous visualized in diseased and healthy stem bases of the resistant cultivar Florence-Aurore at 10, 14 and 21 days after root inoculation. Histological information was used to identify disease relevant tissues and to assist the interpretation of molecular images. Detected mycotoxin compounds secreted by F. graminearum showed a route of stem infection that was consistent with observations made by microscopy. The outer epidermis and vasculature of leaf sheath were, at different disease stages, identified as prominent sites of pathogen migration and wheat protection. Wheat metabolites mapped to these relatively small tissues indicated cell wall strengthening and antifungal activity as direct defenses as well as conservation in the wheat reactions to F. graminearum diseases that affect different plant organs. AP-SMALDI-MS imaging at high spatial resolution is a versatile technique that can be applied to basic and applied aspects of agricultural research. Combining the technology with optical microscopy was found to be a powerful tool to gain in-depth information on almost unknown crop disease. Moreover, the approach allowed studying metabolism at the host-pathogen interface. The results provide important hints to an understanding of the complex spatio-temporal organization of plant resistance. Defense-on-demand responses to pathogen ingress were found, which provide opportunities for future research towards an improved resistance that does not negatively impact yield development in the field by saving plant resources and, moreover, may control different Fusarium diseases.


Regional apparent metabolite concentrations in young adult brain measured by (1)H MR spectroscopy at 3 Tesla.
To quantify and examine the distribution of brain metabolites in normal young adults using single voxel MR spectroscopy at 3 Tesla (T). Short-echo time single-voxel PRESS technique was used to measure the apparent concentration of five metabolites at nine locations in the brains of young adults. Concentrations were estimated by means of an automated fitting method (LCModel) with reference to an unsuppressed water signal and were corrected for T(1) relaxation, T(2) relaxation, and cerebrospinal fluid partial volume. Analysis of variance with Tukey post hoc test was used to evaluate regional variations. Statistically significant differences in regional concentrations were detected for each of the metabolites. The number of significant differences was greatest for total choline, whereas myo-inositol and the sum of glutamine and glutamate had the fewest. Magnitude of variation was greatest for total choline and least for the sum of N-acetyl aspartate and N-acetylaspartylglutamate. In agreement with previous studies at other field strengths, we found heterogeneous distribution of the major spectroscopically measurable brain metabolites. Although the most distinct differences are between tissue types, there is appreciable variation within a tissue type at different locations. The spectra and metabolite concentrations presented should provide a useful reference for both clinical and research MR spectroscopy studies performed at 3T.


Imaging and AI based chromatin biomarkers for diagnosis and therapy evaluation from liquid biopsies.
Multiple genomic and proteomic studies have suggested that peripheral blood mononuclear cells (PBMCs) respond to tumor secretomes and thus could provide possible avenues for tumor prognosis and treatment evaluation. We hypothesized that the chromatin organization of PBMCs obtained from liquid biopsies, which integrates secretome signals with gene expression programs, provides efficient biomarkers to characterize tumor signals and the efficacy of proton therapy in tumor patients. Here, we show that chromatin imaging of PBMCs combined with machine learning methods provides such robust and predictive chromatin biomarkers. We show that such chromatin biomarkers enable the classification of 10 healthy and 10 pan-tumor patients. Furthermore, we extended our pipeline to assess the tumor types and states of 30 tumor patients undergoing (proton) radiation therapy. We show that our pipeline can thereby accurately distinguish between three tumor groups with up to 89% accuracy and enables the monitoring of the treatment effects. Collectively, we show the potential of chromatin biomarkers for cancer diagnostics and therapy evaluation.


The wheat aquaporin gene TaAQP7 confers tolerance to cold stress in transgenic tobacco.
Aquaporin proteins (AQPs) have been shown to be involved in abiotic stress responses. However, the precise role of AQPs, especially in response to cold stress, is not understood in wheat (Triticum aestivum). In the present study, quantitative real time polymerase chain reaction (qRT-PCR) analysis revealed that TaAQP7 expression increased in leaves, but decreased in roots after cold treatment. Expression of TaAQP7 in tobacco plants resulted in increased root elongation and better growth compared with wild-type (WT) plants under cold stress. Moreover, after cold treatment, the transgenic tobacco lines exhibited higher chlorophyll contents, lower levels of malondialdehyde (MDA), and less ion leakage (IL) than WT plants. Thus, expression of TaAQP7 enhanced cold stress tolerance in transgenic tobacco. Taken together, our results suggest that TaAQP7 confers cold stress tolerance by relieving membrane damage in the transgenic plants.


Relation of Serum Albumin Levels to Myocardial Extracellular Volume in Patients With Severe Aortic Stenosis.
Severe aortic stenosis (AS) is often characterized by myocardial interstitial fibrosis. Myocardial interstitial fibrosis, classically measured by magnetic resonance imaging, was also shown to be accurately measured by computed tomography (CT)-derived extracellular volume fraction (ECVF). Serum albumin (SA) level (g/dl) has been shown to correlate with ECVF among patients with heart failure and preserved ejection fraction. Our objective was to evaluate the association between SA and ECVF among patients with severe symptomatic AS. Patients with symptomatic severe AS who were evaluated as candidates for intervention between 2016 and 2018 were enrolled prospectively. All patients underwent precontrast and postcontrast CT for estimating myocardial ECVF. Valid ambulatory SA within 6 weeks of the cardiac CT were obtained and classified as (tertiles) <3.8, 3.8 to 4.19, and ≥4.2 g/dl. Patients with acute systemic illness at the time of the albumin test were excluded. The study included 68 patients, mean age 81 ± 6 years, 53% women. Patients with lower SA were more likely to have chronic renal failure, previous percutaneous coronary interventions, and a reduced functional class. The mean ECVF (%) in the study cohort was 41 ± 12%, significantly higher among the patients in the lower SA level groups (50 ± 12% vs 38 ± 7% vs 33 ± 9% in the <3.8 g/dl, 3.8 to 4.19 g/dl and ≥4.2 g/dl groups respectively, p for trend <0.001). A statistically significant inverse correlation was found between SA levels and ECVF (r -0.7, p <0.001). Multivariable analysis showed significant independent association between low SA and ECVF. In conclusion, the SA level is inversely associated with CT-derived ECVF in patients with severe AS.


Evidence for differential localization of noradrenaline and neuropeptide Y in neuronal storage vesicles isolated from rat vas deferens.
Using the technique of homogenization and subsequent density gradient centrifugation combined with ultrastructural analysis, the subcellular localization of noradrenaline and neuropeptide Y (NPY) was studied in vas deferens of castrated male rats. Noradrenaline showed two peaks in the gradient: one major peak at low density and another at high density. Only one NPY peak was seen, which coincided with the high density peak of noradrenaline. Electron microscopic analysis revealed high proportions of small and large vesicles in the light and heavy fractions, respectively. The present results indicate a differential subcellular localization of noradrenaline and NPY in the noradrenergic nerve endings of vas deferens. Thus, small vesicles seem to contain only noradrenaline, whereas the large vesicles may contain both noradrenaline and NPY.


Comparison of cervicovaginal lavage, cervicovaginal lavage enriched with cervical swab, and vaginal tampon for the detection of HIV-1 RNA and HSV-2 DNA in genital secretions.
We compared the performance of 3 collection methods for cervicovaginal secretions [cervicovaginal lavage (CVL), CVL enriched with a cervical swab (eCVL), and vaginal tampon (VT)] to identify the most reliable method for detection of cervicovaginal HIV-1 and herpes simplex virus type 2 (HSV-2). HIV-1 RNA (Nuclisens EasyQ; BioMerieux, Marcy-l'Etoile, France), HSV-2 DNA (real-time polymerase chain reaction), and microscopic blood and semen traces were detected in samples from 19 HIV-1-HSV-2-coinfected women seen at 4 weekly visits. HIV-1 RNA was detected in 49 (79%) of 62 eCVLs, 41 (61%) of 67 CVLs, and 27 (57%) of 47 VTs. Detection of HIV-1 RNA was higher in eCVL compared with CVL [45/58 (78%) vs. 32/58 (55%); risk ratio 1.41, 95% confidence interval 1.05 to 1.88]. Although more eCVLs were contaminated with microscopic blood (29%) than CVLs (22%) or VTs (7%), detection of HIV-1 RNA remained higher using eCVL compared with CVL (risk ratio 1.43, 95% confidence interval 1.02 to 2.02) in uncontaminated samples. HSV-2 DNA was detected in less than 10% of samples by each method but in 7 (37%) of 19 women overall by 1 or more methods.


UV-induced conformational isomerization and photochemistry of 3-chloro-4-methoxybenzaldehyde in cryogenic inert matrices.
The interest in investigating the structure and reactivity of halogen- and methoxy-substituted benzaldehydes has been motivated by the practical relevance of these substances in various fields, from the fragrance industry to agrochemicals and drug manufacturing. In this study, 3-chloro-4-methoxybenzaldehyde (3CMBA) was investigated by matrix isolation infrared spectroscopy and quantum chemistry calculations. Molecules of the compound were isolated in cryogenic argon and xenon matrices and the conformational composition of the as-deposited materials characterized by infrared spectroscopy. Two nearly degenerated conformers, differing by the orientation of the aldehyde group relatively to the chlorine ring substituent, were successfully trapped into the matrices from the room temperature vapor of the compound, the cis conformer being slightly higher in energy than the trans form. Both conformers belong to the Cs point group and have the methoxy group pointing to the opposite direction of the chlorine atom. The as-deposited matrices of 3CMBA were then irradiated by tunable monochromatic UV-light. Irradiation within the 302-299 nm range induced conversion of the cis into the trans conformer, while irradiation in the 298-296 nm range induced the inverse photoisomerization. Irradiation at higher energy (250 nm) was found to lead to decarbonylation of both conformers. Very interestingly, the observed UV-induced processes were found to take place in an essentially selective way as a function of the irradiation wavelength. In particular, the conformational isomerization reactions of 3CMBA occur with only a residual number of molecules undergoing simultaneous decarbonylation.


Metabolic imaging patterns in posterior cortical atrophy and Lewy body dementia.
To study the imaging patterns of Posterior cortical atrophy (PCA) and Dementia with Lewy bodies (DLB) on fluoro-deoxyglucose positron emission tomography computed tomography ([F]FDG PET/CT), identify areas of overlap and differences and to develop a prediction model to assist in diagnosis using univariate and multivariate analysis. A retrospective analysis of 72 patients clinically suspected of having posterior dementia was done. All patients underwent [FF]FDG PET/CT of the brain and dopamine transporter imaging with [[Tc]TRODAT-1 SPECT scan on separate days. The patients were divided into PCA with normal TRODAT uptake (n=34) and DLB with abnormal TRODAT uptake (n=38). The FDG PET/CT uptake patterns were recorded and areas of significant hypometabolism by z score analysis were considered as abnormal. Receiver operator characteristics (ROC) curve analysis was used to determine cutoff z scores and binary logistic regression analysis was used to determine the Odds ratio of being in the predicted groups. Significantly hypometabolism was found in parieto-temporo-occipital association cortices and cingulate cortices in PCA patients. DLB patients showed significantly reduced uptake in the visual cortex. No significant difference was found between z score of occipital association cortex which showed hypometabolism in both groups. The cut-off z-score values derived from the ROC curve analysis were as follows- parietal association (cut-off-3, sensitivity-65.6%, specificity - 68.7%), temporal association (cut-off-2, sensitivity-78%, specificity-75%) and posterior cingulate (cut-off-0.5, sensitivity-93.7%, specificity-40.6%), their respective Odds ratio (with 95% confidence interval) for being in the PCA group as derived from univariate logistic regression were 3.66 (1.30-10.32), 10.71 (3.36-34.13) and 7.85 (1.57-39.17). The cut-off z score of primary visual cortex as derived from ROC curve was zero with sensitivity of 87.5%, specificity of 71.9%, and the Odds ratio for being the in the DLB group was 24.7 with 95% confidence interval of 5.99-101.85. [F]FDG PET may be useful as a non-invasive diagnostic modality in differentiating the two posterior cortical dementias, despite significant overlap. Primary visual cortical hypometabolism can serve as an independent diagnostic marker for DLB, even in the absence of TRODAT imaging.


Stress physiology as a predictor of survival in Galapagos marine iguanas.
Although glucocorticoid hormones are considered important physiological regulators for surviving adverse environmental stimuli (stressors), evidence for such a role is sparse and usually extrapolated from glucocorticoid effects under laboratory, short-term and/or non-emergency conditions. Galápagos marine iguanas (Amblyrhynchus cristatus) provide an excellent model for determining the ultimate function of a glucocorticoid response because susceptibility to starvation induced by El Niño conditions is essentially their only major natural stressor. In a prospective study, we captured 98 adult male marine iguanas and assessed four major components of their glucocorticoid response: baseline corticosterone titres; corticosterone responses to acute stressors (capture and handling); the maximal capacity to secrete corticosterone (via adrenocorticotropin injection); and the ability to terminate corticosterone responses (negative feedback). Several months after collecting initial measurements, weak El Niño conditions affected the Galápagos and 23 iguanas died. The dead iguanas were typified by a reduced efficacy of negative feedback (i.e. poorer post-stress suppression of corticosterone release) compared with surviving iguanas. We found no prior differences between dead and alive iguanas in baseline corticosterone concentrations, responses to acute stressors, nor in capacity to respond. These data suggest that a greater ability to terminate a stress response conferred a survival advantage during starvation.


Role of Cytokines in the Pathogenesis of Visceral Leishmaniasis.
Infection in humans with Leishmania manifests into a spectrum of diseases. The manifestations of the disease depend on the resultant evasion of the parasite to immune responses namely macrophages, which is an exclusive host of leishmania. The B cells valiantly mount antibody responses, however to no avail as the Leishmania parasites occupy the intracellular niches of the macrophages. Extensive studies have been documented on the role of cell-mediated immunity (CMI) in protection and counter survival strategies of the parasites leading to down-regulation of CMI. The present review attempts to discuss the cytokines in progression or resolution of visceral form of leishmaniasis or kala-azar, predominantly affecting the Indian subcontinent. The components/cytokine(s) responsible for the regulation of the critical balance of Th1/Th2/Th9/Th17/Treg cells has been discussed in the perspective. Therefore, any strategy involving the treatment of VL needs to consider the balance and regulation of CD4+ T cell function.


Electrical stimulation of the hypoglossal nerve: a potential therapy.
Obstructive sleep apnea is characterized by recurrent episodes of pharyngeal collapse, which result from a decrease in pharyngeal dilator muscle tone. The genioglossus is a major pharyngeal dilator that maintains airway patency during sleep. Early studies in animal and humans have demonstrated that electrical stimulation of this muscle reduces pharyngeal collapsibility, increases airflow, and mitigates obstructive sleep apnea. These findings impelled the development of fully implantable hypoglossal nerve stimulating systems (HGNS), for which feasibility trial results are now available. These pilot studies have confirmed that hypoglossal nerve stimulation can prevent pharyngeal collapse without arousing patients from sleep. Potentially, a substantial segment of the patient population with obstructive sleep apnea can be treated with this novel approach. Furthermore, the feasibility trial findings suggest that the therapeutic potential of HGNS can be optimized by selecting patients judiciously, titrating the stimulus intensity optimally, and characterizing the underlying function and anatomy of the pharynx. These strategies are currently being examined in ongoing pivotal trials of HGNS.


Olfactory heterogeneity in LRRK2 related Parkinsonism.
LRRK2 mutations can cause familial and sporadic Parkinson's disease (PD) with Lewy-body pathology at post-mortem. Studies of olfaction in LRRK2 are sparse and incongruent. We applied a previously validated translation of the 16 item smell identification test from Sniffin' Sticks (SS-16) to 14 parkinsonian carriers of heterozygous G2019S LRRK2 mutation and compared with 106 PD patients and 118 healthy controls. The mean SS-16 score in LRRK2 was higher than in PD (p < 0.001, 95% CI for β = -4.7 to -1.7) and lower than in controls (p = 0.007, 95% CI for β = +0.6 to +3.6). In the LRRK2 group, subjects with low scores had significantly more dyskinesia. They also had younger age of onset, longer disease duration, and reported less frequently a family history of PD, but none of these other differences reached significance. Odor identification is diminished in LRRK2 parkinsonism but not to the same extent as in idiopathic PD.


Subjective states induced by intracranial electrical stimulation matches the cytoarchitectonic organization of the human insula.
Functions of the human insula have been explored extensively with neuroimaging methods and intracranial electrical stimulation studies that have highlighted a functional segregation across its subregions. A recently developed cytoarchitectonic map of the human insula has also segregated this brain region into various areas. Our knowledge of the functional organization of this brain region at the level of these fine-parceled microstructural areas remains only partially understood. We address this gap of knowledge by applying a multimodal approach linking direct electrical stimulation and task-evoked intracranial EEG recordings with microstructural subdivisions of the human insular cortex. In 17 neurosurgical patients with 142 implanted electrodes, stimulation of 40 % of the sites induced a reportable change in the conscious experience of the subjects in visceral/autonomic, anxiety, taste/olfactory, pain/temperature as well as somatosensory domains. These subjective responses showed a topographical allocation to microstructural areas defined by probabilistic cytoarchitectonic parcellation maps of the human insula. We found the pain and thermal responses to be located in areas lg2/ld2, while non-painful/non-thermal somatosensory responses corresponded to area ld3 and visceroceptive responses to area Id6. Lastly, the stimulation of area Id7 in the dorsal anterior insula, failed to induce reportable changes to subjective experience even though intracranial EEG recordings from this region captured significant time-locked high-frequency activity (HFA). Our results provide a multimodal map of functional subdivisions within the human insular cortex at the individual brain basis and characterize their anatomical association with fine-grained cytoarchitectonic parcellations of this brain structure.


Multiple risk factors during pregnancy in South Africa: the need for a horizontal approach to perinatal care.
South African children's long-term health and well-being is jeopardized during their mothers' pregnancies by the intersecting epidemics of HIV, alcohol use, low birth weight (LBW; <2,500 g) related to poor nutrition, and depressed mood. This research examines these overlapping risk factors among 1,145 pregnant Xhosa women living in 24 township neighborhoods in Cape Town, South Africa. Results revealed that 66 % of pregnant women experienced at least one risk factor. In descending order of prevalence, 37 % reported depressed mood, 29 % were HIV+, 25 % used alcohol prior to knowing that they were pregnant, and 15 % had a previous childbirth with a LBW infant. Approximately 27 % of women had more than one risk factor: depressed mood was significantly associated with alcohol use and LBW, with a trend to significance with HIV+. In addition, alcohol use was significantly related to HIV+. These results suggest the importance of intervening across multiple risks to maternal and child health, and particularly with depression and alcohol use, to positively impact multiple maternal and infant outcomes.


Social-emotional functioning in pediatric hydrocephalus: comparison of the Hydrocephalus Outcome Questionnaire to the Behavior Assessment System for Children.
Hydrocephalus can impact all areas of health, including physical, cognitive, and social-emotional functioning. The social-emotional health of children who have had surgery for hydrocephalus is not well characterized. In this study, the authors sought to examine social-emotional functioning using the Behavior Assessment System for Children, Third Edition (BASC-3) and the Hydrocephalus Outcome Questionnaire (HOQ) in 66 children aged 5 to 17 years. Caregivers of pediatric patients with hydrocephalus completed the BASC-3 and the HOQ. BASC-3 internalizing, externalizing, and executive functioning caregiver-reported scores were compared with the BASC-3 normative sample using one-sample t-tests to evaluate overall social-emotional functioning. BASC-3 scores were correlated with the social-emotional domain of the HOQ using Pearson's r to determine if the HOQ accurately captured the social-emotional functioning of children with hydrocephalus in a neurosurgery setting. BASC-3 and HOQ scores of children with different etiologies of hydrocephalus were compared using the Kruskal-Wallis one-way analysis of variance to determine if differences existed between the following etiologies: intraventricular hemorrhage secondary to prematurity, myelomeningocele, communicating congenital hydrocephalus, aqueductal stenosis, or other. Children with hydrocephalus of all etiologies had more difficulties with social-emotional functioning compared with normative populations. Children with different hydrocephalus etiologies differed in executive functioning and overall HOQ scores but not in internalizing symptoms, externalizing symptoms, or social-emotional HOQ scores. The social-emotional domain of the HOQ correlated more strongly with the BASC-3 than did the physical and cognitive domains. These results have provided evidence that children who have had surgery for hydrocephalus may be at increased risk of social-emotional and behavioral difficulties, but etiology may not be particularly helpful in predicting what kinds or degree of difficulty. The results of this study also support the convergent and divergent validity of the social-emotional domain of the HOQ.


[Lymphocyte metabolism in patients with acute pancreatitis with different genotypes of GSTM1 and GSTT1 genes].
In this study, we have investigated correlation between enzymatic activity of NAD(P)-dependent dehydrogenases of lymphocytes and polymorphic variants of glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) genes in the group of unrelated patients with acute pancreatitis in comparison with healthy Russians from Krasnoyarsk. Thus, genotype GSTM1 0/0 is the marker of predisposition to the acute pancreatitis, wheras polymorphism of the GSTT1 gene is not involved in the development of the pancreatitis, at least in our group. The bioluminescence analysis showed statistically significant decrease of the levels of G3PD, NAD(+)MDH and the increase of NADH(+)LDH, NAD(+)GDH, NADH(+)GDH in lymphocytes of pancreatic group. Development of pancreatitis in patients with different genotypes GSTM1 and GSTT1 genes showed the rearrangement of the basic intracellular processes: dominance of a plastic metabolism in the patients with GSTM1--deletions and predominance of energetic processes at GSTT1 0 - pancreatitis.


Maternal risk for down syndrome and polymorphisms in the promoter region of the DNMT3B gene: a case-control study.
Epigenetic changes leading to improper methylation of the pericentromeric region of chromosome 21 may contribute to the nondisjunction of this chromosome. Polymorphisms in the DNA Methyltransferase 3B (DNMT3B) gene, one of the crucial gene of the folate metabolism, affects the activity of the enzyme and increases the susceptibility of nondisjunction in mothers of Down syndrome children (MDS). Considering this hypothesis we investigated the association of single nucleotide polymorphisms in the promoter region of the DNMT3B gene (rs1569686 -579G>T; rs2424913 -149C>T) with a predisposition of mothers to deliver a Down syndrome (DS) child. The study was performed on DNA samples from 150 MDS and 172 control mothers. Transmission disequilibrium tests were performed on 103 DS trio families. Genotyping was done using a polymerase chain reaction-restriction fragment length polymorphism method. With respect to the single nucleotide polymorphisms studied, no significant difference was observed in the genotypes and alleles frequency distributions between MDS and control mothers. The frequency of the DNMT3B-579G allele was, respectively, 0.34 in MDS and 0.33 in control mothers whereas the frequency of the DNMT3B-149C allele was respectively 0.31 in MDS and 0.26 in control mothers. No significant deviation in genotypic combinations as well as in transmission disequilibrium tests analysis was observed. However, a strong linkage disequilibrium was observed with significant differences in the distribution of G-T and G-C haplotypes among case and control mothers. Although the above studied polymorphisms of DNMT3B may not be an independent risk factor it might be possible that certain allelic combinations (G-T) are. This finding suggests that DNMT3B might be a maternal risk factor for DS in our Indian cohort. Replication studies are required to confirm these findings.


Space-time data-driven modeling of precipitation-induced shallow landslides in South Tyrol, Italy.
Shallow landslides represent potentially damaging processes in mountain areas worldwide. These geomorphic processes are usually caused by an interplay of predisposing, preparatory, and triggering environmental factors. At regional scales, data-driven methods have been used to model shallow landslides by addressing the spatial and temporal components separately. So far, few studies have explored the integration of space and time for landslide prediction. This research leverages generalized additive mixed models to develop an integrated approach to model shallow landslides in space and time. We built upon data on precipitation-induced landslide records from 2000 to 2020 in South Tyrol, Italy (7400 km2). The Slope Unit-based model predicts landslide occurrence as a function of static and dynamic factors while seasonal effects are incorporated. The model also accounts for spatial and temporal biases inherent in the underlying landslide data. We validated the resulting predictions through a suite of cross-validation techniques, obtaining consistent performance scores above 0.85. The analyses revealed that the best-performing model combines static ground conditions and two precipitation time windows: a short-term cumulative precipitation of 2 days before the landslide event and a medium-term cumulative precipitation of 14 days. We demonstrated the model's predictive capabilities by predicting the dynamic landslide probabilities over historical data associated with a heavy precipitation event on August 4th and August 5th, 2016, and hypothetical non-spatially explicit precipitation (what-if) scenarios. The novel approach shows the potential to integrate static and dynamic landslide factors for large areas, accounting for the underlying data structure and data limitations.


A Systematic Review of Rehabilitation Exercises to Progressively Load the Gluteus Medius.
Gluteus medius rehabilitation is of critical importance given its role in pelvic and lower limb stability, and the known link between gluteus medius weakness and many lower limb conditions. To systematically review the literature and present an evidence-based graduated series of exercises to progressively load gluteus medius. A systematic literature search was conducted in January 2016 to identify studies reporting gluteus medius muscle activity as a percentage of maximal volitional isometric contraction (MVIC), during rehabilitation exercises. Studies that investigated injury free participants were included. No restrictions were placed on the type or mode of exercise, though exercises that could not be accurately replicated or performed within an independent setting were excluded. Studies that did not normalize electromyographic activity to a side lying MVIC were excluded. Exercises were stratified based on exercise type and %MVIC: low (0% to 20%), moderate (21% to 40%), high (41% to 60%), and very high (> 61%). 20 studies were included in this review, reporting outcomes in 33 exercises (and a range of variations of the same exercise). Prone, quadruped, and bilateral bridge exercises generally produced low or moderate load. Specific hip abduction/rotation exercises were reported as moderate, high, or very high load. Unilateral stance exercises in the presence of contralateral limb movement were often high or very high load activities, while high variability existed across a range of functional weight-bearing exercises. This review outlined a series of exercises commonly employed in a rehabilitation setting, stratified based on exercise type and the magnitude of gluteus medius muscular activation. This will assist clinicians in tailoring gluteus medius loading regimens to patients, from the early postoperative through to later stages of rehabilitation.


Chiari I malformation and pregnancy: a comprehensive review of the literature to address common questions and to guide management.
The optimal management of Chiari I malformation during pregnancy remains uncertain. Labor contractions, which increase intracranial pressure, and neuraxial anesthesia both carry the theoretical risk of brainstem herniation given the altered CSF dynamics inherent to the condition. Mode of delivery and planned anesthesia, therefore, require forethought to avoid potentially life-threatening complications. Since the assumed potential risks are significant, we seek to systematically review published literature regarding Chiari I malformation in pregnancy and, therefore, to establish a best practice recommendation based on available evidence. The English-language literature was systematically reviewed from 1991 to 2018 according to PRISMA guidelines to assess all pregnancies reported in patients with Chiari I malformation. After analysis, a total of 34 patients and 35 deliveries were included in this investigation. Additionally, a single case from our institutional experience is presented for illustrative purposes but not included in the statistical analysis. No instances of brain herniation during pregnancy in patients with Chiari I malformation were reported. Cesarean deliveries (51%) and vaginal deliveries (49%) under neuraxial blockade and general anesthesia were both reported as safe and suitable modes of delivery. Across all publications, only one patient experienced a worsening of neurologic symptoms, which was only later discovered to be the result of a previously undiagnosed Chiari I malformation. Several patients underwent decompressive suboccipital craniectomy to treat the Chiari I malformation during the preconception period (31%), during pregnancy (3%), and after birth (6%). Specific data regarding maternal management were not reported for a large number (21) of these patients (60%). Aside from one abortion in our own institutional experience, there was no report of any therapeutic abortion or of adverse fetal outcome. Although devastating maternal complications are frequently feared, very few adverse outcomes have ever been reported in pregnant patients with a Chiari I malformation. The available evidence is, however, rather limited. Based on our survey of available data, we recommend vaginal delivery under neuraxial blockade for truly asymptomatic patients. Furthermore, based on our own experience and physiological conceptual considerations, we recommend limiting maternal Valsalva efforts either via Cesarean delivery under regional or general anesthesia or by choosing assisted vaginal delivery under neuraxial blockade. There is no compelling reason to offer suboccipital decompression for Chiari I malformation during pregnancy. For patients with significant neurologic symptoms prior to conception, decompression prior to pregnancy should be considered.


Evaluation of the 'Best Start' breast-feeding education program.
To compare the frequency of breast-feeding before and after the "Best Start" breast-feeding educational program was implemented. Preintervention-postintervention trial. A women's health center and children's hospital serving a low-income population. Ninety mother-infant pairs of whom the infants were born between January 2 and February 28, 1993, compared with 90 mother-infant pairs of whom the infants were born between January 2 and February 28, 1994. Best Start, a breast-feeding education program, was presented to health professionals and clerical staff of the Women's Health Center. At the first prenatal visit, a woman is asked, "What do you know about breast-feeding?" instead of "Are you going to breast-feed or bottle-feed this baby?" The program elicits and acknowledges the mother's concerns and then educates her about the benefits of breast-feeding. This is repeated at each prenatal visit. Sociodemographic data and breast-feeding rates of preintervention groups were compared with those of postintervention groups. Both hospital and out-patient records were reviewed. In 1993, 13 (15%) of 86 mothers breast-fed at hospital discharge compared with 25 (31%) of 81 in 1994 (P < .03). At the 2-week clinic visit, 11 (13%) of 86 were still breast-feeding in 1993 compared with 17 (21%) of 81 in 1994 (P > .20). The impact on mothers aged 19 years or less was particularly marked, with a tripling of the breast-feeding rate at hospital discharge from 11% (2/18) in 1993 to 37% (10/27) in 1994. This simple, low-cost educational intervention with a change in the manner of presentation significantly improved the breast-feeding rates of this low-income population.


Physiological covalent regulation of rat liver branched-chain alpha-ketoacid dehydrogenase.
A radiochemical assay was developed for measuring branched-chain alpha-ketoacid dehydrogenase activity of Triton X-100 extracts of freeze-clamped rat liver. The proportion of active (dephosphorylated) enzyme was determined by measuring enzyme activities before and after activation of the complex with a broad-specificity phosphoprotein phosphatase. Hepatic branched-chain alpha-ketoacid dehydrogenase activity in normal male Wistar rats was 97% active but decreased to 33% active after 2 days on low-protein (8%) diet and to 13% active after 4 days on the same diet. Restricting protein intake of lean and obese female Zucker rats also caused inactivation of hepatic branched-chain alpha-ketoacid dehydrogenase complex. Essentially all of the enzyme was in the active state in rats maintained for 14 days on either 30 or 50% protein diets. This was also the case for rats maintained on a commercial chow diet (minimum 23% protein). However, maintaining rats on 20, 8, and 0% protein diets decreased the percentage of the active form of the enzyme to 58, 10, and 7% of the total, respectively. Fasting of chow-fed rats for 48 h had no effect on the activity state of hepatic branched-chain alpha-ketoacid dehydrogenase, i.e., 93% of the enzyme remained in the active state compared to 97% for chow-fed rats. However, hepatic enzyme of rats maintained on 8% protein diet was 10% active before starvation and 83% active after 2 days of starvation. Thus, dietary protein deficiency results in inactivation of hepatic branched-chain alpha-ketoacid dehydrogenase complex, presumably as a consequence of low hepatic levels of branched-chain alpha-ketoacids, established inhibitors of branched-chain alpha-ketoacid dehydrogenase kinase. With rats fed a low-protein diet and subsequently starved, inhibition of branched-chain alpha-ketoacid dehydrogenase kinase by branched-chain alpha-ketoacids generated as a consequence of endogenous proteolysis most likely promotes the greater branched-chain alpha-ketoacid dehydrogenase activity state.


Early repolarization pattern: a marker of increased risk in patients with catecholaminergic polymorphic ventricular tachycardia.
The early repolarization pattern (ERP) has been shown to be associated with arrhythmias in patients with short QT syndrome, Brugada syndrome, and ischaemic heart disease. Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited arrhythmia syndrome and related to malignant ventricular tachyarrhythmias in a structurally normal heart. The aim of this study was to evaluate the prevalence of ERP and clinical events in patients with CPVT. Digitalized resting 12-lead ECGs of patients were analysed for ERP and for repolarization markers (QT and Tpeak-Tend interval). The ERP was diagnosed as 'notching' or 'slurring' at the terminal portion of QRS with ≥0.1 mV elevation in at least two consecutive inferior (II, III, aVF) and/or lateral leads (V4-V6, I, aVL). Among 51 CPVT patients (mean age 36 ± 15 years, 11 males), the ERP was present in 23 (45%): strictly in the inferior leads in 9 (18%) patients, in the lateral leads in 9 (18%) patients, and in infero-lateral leads in 5 (10%) patients. All patients with ERP were symptomatic at presentation (23 of 23 patients with ERP vs. 19 of 28 patients without ERP, P = 0.003). Syncope was also more frequent in patients with ERP (18 of 23 patients with ERP vs. 11 of 28 patients without ERP, P = 0.005). A pathologic ERP is present in an unexpected large proportion (45%) of patients and is associated with an increased frequency of syncope. In patients with unexplained syncope and ERP at baseline, exercise testing should be performed to detect CPVT.


A comparison of immunocytochemical markers to identify bipolar cell types in human and monkey retina.
As more human retinas affected with genetic or immune-based diseases become available for morphological analysis, it is important to identify immunocytochemical markers for specific subtypes of retinal neurons. In this study, we have focused on bipolar cell markers in central retina. We have done single and double labeling using several antisera previously utilized in macaque monkey or human retinal studies and two new antisera (1) to correlate combinations of antisera labeling with morphological types of bipolar cells in human retina, and (2) to compare human labeling patterns with those in monkey retina. Human bipolar cells showed a wide range of labeling patterns with at least ten different bipolar cell types identified from their anatomy and marker content. Many bipolar cell bodies in the outer part of the inner nuclear layer contained combinations of protein kinase C alpha (PKC alpha), Islet-1, glycine, and Go alpha. Bipolar cells labeled with these markers had axons terminating in the inner half of the inner plexiform layer (IPL), consistent with ON bipolar cells. Bipolar cell bodies adjacent to the amacrine cells and with axons in the outer half of the IPL contained combinations of recoverin, glutamate transporter-1, and PKC beta, or CD15 and calbindin. Bipolar cells labeled with these markers were presumed OFF bipolar cells. Calcium-binding protein 5 (CaB5) labeled both putative ON and OFF bipolar cells. Using this cell labeling as a criteria, most cell bodies close to the horizontal cells were ON bipolar cells and almost all bipolar cells adjacent to the amacrine cells were OFF with a band in the middle 2-3 cell bodies thick containing intermixed ON and OFF bipolar cells. Differences were found between human and monkey bipolar cell types labeled by calbindin, CaB5, and CD15. Two new types were identified. One was morphologically similar to the DB3, but labeled for CD15 and CaB5. The other had a calbindin-labeled cell body adjacent to the horizontal cell bodies, but did not contain any accepted ON markers. These results support the use of macaque monkey retina as a model for human, but caution against the assumption that all labeling patterns are identical in the two primates.


The optimal DFT approach in DP4 NMR structure analysis - pushing the limits of relative configuration elucidation.
What computational methods should be used to achieve the most reliable result in computational structure elucidation? A study on the effect of quality and quantity of geometries on computational NMR structure elucidation performance is reported. Semi-empirical, HF and DFT methods were explored, and B3LYP optimized geometries in combination with mPW1PW91 shifts and M06-2X conformer energies was found to be best. The required number of conformers considered has also been investigated, as well as several methods for the reduction of this number. Clear guidelines for the best computational NMR structure elucidation methods for different levels of available computing power are provided.


Ultrafast molecule separation through layered WS(2) nanosheet membranes.
Two-dimensional layered materials have joined in the family of size-selective separation membranes recently. Here, chemically exfoliated tungsten disulfide (WS2) nanosheets are assembled into lamellar thin films and explored as an ultrafast separation membrane for small molecules with size of about 3 nm. Layered WS2 membranes exhibit 5- and 2-fold enhancement in water permeance of graphene oxide membranes and MoS2 laminar membranes with similar rejection, respectively. To further increase the water permeance, ultrathin nanostrands are used as templates to generate more fluidic channel networks in the WS2 membrane. The water permeation behavior and separation performance in the pressure loading-unloading process reveal that the channels created by the ultrathin nanostrands are cracked under high pressure and result in a further 2-fold increase of the flux without significantly degrading the rejection for 3 nm molecules. This is supported by finite-element-based mechanical simulation. These layered WS2 membranes demonstrate up to 2 orders of magnitude higher separation performance than that of commercial membranes with similar rejections and hold the promising potential for water purification.


Comparison of five automated serum and whole blood folate assays.
Serum and whole blood folate measurements are used to establish folate deficiency. Most methods used in clinical laboratories are automated, nonistopic methods that use folate-binding protein. Linearity, imprecision, and method comparison studies, including serum and whole blood hemolysates, were performed with the Access, Advia Centaur, ARCHITECT i2000, Elecsys 2010, and IMMULITE 2000 methods. The QuantaPhase II radioassay served as the comparison method. (Proprietary information is given in the text.) The Access and IMMULITE 2000 methods had higher systematic errors in linearity studies than the other 3 methods. The imprecision of all methods was acceptable (coefficient of variation, < 10%) even at low folate concentrations with the exception of the Elecsys 2010 (coefficient of variation, 16%). Method comparison studies using serum samples revealed calibration differences between the Access and Elecsys 2010 methods and the comparison method. Method comparison studies using whole blood samples showed poorer agreement between each of the automated methods and the comparison method than was seen with serum samples. The ARCHITECT i2000 folate assay demonstrated the best analytic performance. The poor agreement seen with whole blood hemolysates likely is due to calibration differences and differences in hemolysate preparation conditions.


Distinct malaria parasite sporozoites reveal transcriptional changes that cause differential tissue infection competence in the mosquito vector and mammalian host.
The malaria parasite sporozoite transmission stage develops and differentiates within parasite oocysts on the Anopheles mosquito midgut. Successful inoculation of the parasite into a mammalian host is critically dependent on the sporozoite's ability to first infect the mosquito salivary glands. Remarkable changes in tissue infection competence are observed as the sporozoites transit from the midgut oocysts to the salivary glands. Our microarray analysis shows that compared to oocyst sporozoites, salivary gland sporozoites upregulate expression of at least 124 unique genes. Conversely, oocyst sporozoites show upregulation of at least 47 genes (upregulated in oocyst sporozoites [UOS genes]) before they infect the salivary glands. Targeted gene deletion of UOS3, encoding a putative transmembrane protein with a thrombospondin repeat that localizes to the sporozoite secretory organelles, rendered oocyst sporozoites unable to infect the mosquito salivary glands but maintained the parasites' liver infection competence. This phenotype demonstrates the significance of differential UOS expression. Thus, the UIS-UOS gene classification provides a framework to elucidate the infectivity and transmission success of Plasmodium sporozoites on a whole-genome scale. Genes identified herein might represent targets for vector-based transmission blocking strategies (UOS genes), as well as strategies that prevent mammalian host infection (UIS genes).


Early versus delayed endocrine treatment of T2-T3 pN1-3 M0 prostate cancer without local treatment of the primary tumour: final results of European Organisation for the Research and Treatment of Cancer protocol 30846 after 13 years of follow-up (a randomised controlled trial).
The timing of endocrine treatment (ET) for prostate cancer (PCa) remains controversial. The issue is addressed in European Organisation for the Research and Treatment of Cancer (EORTC) protocol 30846 for patients with lymph node-positive (pN1-3) cancer without local treatment of the primary tumour. To evaluate the effect of early versus delayed treatment in pN1-3 PCa. Two hundred thirty-four patients with histologically proven PCa and nodal metastases (pN1-3) were randomized to immediate versus delayed ET without treatment of the primary tumour. ET consisted of a depot luteinising hormone-releasing hormone (LHRH) agonist and 1 mo of an anti-androgen or surgical castration. The trial's main objective was to show non-inferiority of delayed ET to immediate ET by ruling out a hazard ratio (HR) of 1.50 for overall survival (OS), with 85% power at one-sided alpha=5%. All but three patients were treated as randomized. The median follow-up is 13 yr. The median protocol treatment duration was 2.7 yr in the delayed and 3.2 yr in the immediate ET groups. Overall, 193 patients (82.5%) have died (97 on delayed ET and 96 on immediate ET), 59.4% of them as a result of PCa. The intention-to-treat analysis shows a 22% increase in the hazard of death of those randomized to delayed treatment (HR=1.22, 95% confidence interval [CI]: 0.92, 1.62). The difference is not statistically significant, but non-inferiority is also not proved. The median OS on immediate ET is 7.6 yr (95% CI, 6.3-8.3 yr) versus 6.1 yr (95% CI, 5.7-7.3 yr) in the delayed ET group. The 10-yr cumulative incidence of death resulting from PCa was 55.6% in the delayed ET group versus 52.1% with immediate ET group. Similar conclusions hold for PCa-specific survival. After 13 years of follow-up, survival or PCa-specific survival between immediate and delayed ET appear similar, but the trial is underpowered to reach its goal of showing non-inferiority.


Data compression and regression through local principal curves and surfaces.
We consider principal curves and surfaces in the context of multivariate regression modelling. For predictor spaces featuring complex dependency patterns between the involved variables, the intrinsic dimensionality of the data tends to be very small due to the high redundancy induced by the dependencies. In situations of this type, it is useful to approximate the high-dimensional predictor space through a low-dimensional manifold (i.e., a curve or a surface), and use the projections onto the manifold as compressed predictors in the regression problem. In the case that the intrinsic dimensionality of the predictor space equals one, we use the local principal curve algorithm for the the compression step. We provide a novel algorithm which extends this idea to local principal surfaces, thus covering cases of an intrinsic dimensionality equal to two, which is in principle extendible to manifolds of arbitrary dimension. We motivate and apply the novel techniques using astrophysical and oceanographic data examples.