Kinetics of the benzyl + HO2 and benzoxyl + OH barrierless association reactions: fate of the benzyl hydroperoxide adduct under combustion and atmospheric conditions.
Radical-radical association reactions are challenging to address theoretically due to difficulties finding the bottleneck that variationally minimizes the reactive flux. For this purpose, the variable reaction coordinate (VRC) formulation of the variational transition state theory (VTST) represents an appropriate tool. In this work, we revisited the kinetics of two radical-radical association reactions of importance in combustion modelling and poly-aromatic hydrocarbon (PAH) chemistry by performing VRC calculations: benzyl + HO2 and benzoxyl + OH, both forming the adduct benzyl hydroperoxide. Our calculated rate constants are significantly lower than those previously reported based on VTST calculations, which results from a more efficient minimization of the reactive flux through the bottleneck achieved by the VRC formulation. Both reactions show different trends in the variation of their rate constants with temperature. We observed that if the pair of single occupied molecular orbitals (SOMOs) of the associating radicals show a similar nature, i.e. similar character, and thereby a small energy gap, a highly stabilized transition state structure is formed as the result of a very efficient SOMO-SOMO overlap, which may cancel out the free energy bottleneck for the formation of the adduct and result in large rate constants with a negative temperature dependence. This is the case of the benzoxyl and OH radical pair, whose SOMOs show O2p nature with an energy gap of 20.2 kcal mol-1. On the other hand, the benzyl and HO2 radical pair shows lower rate constants with a positive temperature dependence due to the larger difference between both SOMOs (a 28.9 kcal mol-1 energy gap) as a consequence of the contribution of the multiple resonance structures of the benzyl radical. The reverse dissociation rate constants were also calculated using multi-structural torsional anharmonicity partition functions, which were not included in previous work, and the results show a much slower dissociation of benzyl hydroperoxide. Our work may help to improve kinetic models of interest in combustion and PAH formation, as well as to gain further understanding of radical-radical association reactions, which are ubiquitous in different environments.


EEG dynamics reflect the distinct cognitive process of optic problem solving.
This study explores the changes in electroencephalographic (EEG) activity associated with the performance of solving an optics maze problem. College students (N = 37) were instructed to construct three solutions to the optical maze in a Web-based learning environment, which required some knowledge of physics. The subjects put forth their best effort to minimize the number of convexes and mirrors needed to guide the image of an object from the entrance to the exit of the maze. This study examines EEG changes in different frequency bands accompanying varying demands on the cognitive process of providing solutions. Results showed that the mean power of θ, α1, α2, and β1 significantly increased as the number of convexes and mirrors used by the students decreased from solution 1 to 3. Moreover, the mean power of θ and α1 significantly increased when the participants constructed their personal optimal solution (the least total number of mirrors and lens used by students) compared to their non-personal optimal solution. In conclusion, the spectral power of frontal, frontal midline and posterior theta, posterior alpha, and temporal beta increased predominantly as the task demands and task performance increased.


The transcriptome profile of RPE cells by the fullerenol against hydrogen peroxide stress.
Age-related macular degeneration (AMD) causes central vision impairment with increased incidence. In the pathogenesis of AMD, reactive oxygen species (ROS) are associated with RPE cell apoptosis. H2O2 is an oxidative toxicant and is used to establish the AMD in vitro model. However, the mechanisms of ROS in H2O2-induced AMD are still unclear. Fullerenol, a promising antioxidant of nanomaterials, protects RPE cells from ROS attack. In addition to working as a scavenger, little is known about the antioxidant mechanism of fullerenol in RPE cells. In this study, transcriptome sequencing was performed to examine the global changes in mRNA transcripts induced by H2O2 in human ARPE-19 cells. Moreover, we comprehensively investigated the protective effects of fullerenol against H2O2-induced oxidative injury by RNA sequencing. Gene Ontology enrichment analysis showed that those pathways related to the release of positive regulation of DNA-templated transcription and negative regulation of apoptotic process were affected. Finally, we found that 12 hub genes were related to the oxidative-protection function of fullerenol. In summary, H2O2 affected these hub genes and signaling pathways to regulate the senescence of RPE cells. Moreover, fullerenol is a potent nanomaterial that protects the RPE and would be a promising approach for AMD prevention.


Construction of a risk stratification model integrating ctDNA to predict response and survival in neoadjuvant-treated breast cancer.
The pathological complete response (pCR) to neoadjuvant chemotherapy (NAC) of breast cancer is closely related to a better prognosis. However, there are no reliable indicators to accurately identify which patients will achieve pCR before surgery, and a model for predicting pCR to NAC is required. A total of 269 breast cancer patients in Shandong Cancer Hospital and Liaocheng People's Hospital receiving anthracycline and taxane-based NAC were prospectively enrolled. Expression profiling using a 457 cancer-related gene sequencing panel (DNA sequencing) covering genes recurrently mutated in breast cancer was carried out on 243 formalin-fixed paraffin-embedded tumor biopsies samples before NAC from 243 patients. The unique personalized panel of nine individual somatic mutation genes from the constructed model was used to detect and analyze ctDNA on 216 blood samples. Blood samples were collected at indicated time points including before chemotherapy initiation, after the 1st NAC and before the 2nd NAC cycle, during intermediate evaluation, and prior to surgery. In this study, we characterized the value of gene profile mutation and circulating tumor DNA (ctDNA) in combination with clinical characteristics in the prediction of pCR before surgery and investigated the prognostic prediction. The median follow-up time for survival analysis was 898 days. Firstly, we constructed a predictive NAC response model including five single nucleotide variant (SNV) mutations (TP53, SETBP1, PIK3CA, NOTCH4 and MSH2) and four copy number variation (CNV) mutations (FOXP1-gain, EGFR-gain, IL7R-gain, and NFKB1A-gain) in the breast tumor, combined with three clinical factors (luminal A, Her2 and Ki67 status). The tumor prediction model showed good discrimination of chemotherapy sensitivity for pCR and non-pCR with an AUC of 0.871 (95% CI, 0.797-0.927) in the training set, 0.771 (95% CI, 0.649-0.883) in the test set, and 0.726 (95% CI, 0.556-0.865) in an extra test set. This tumor prediction model can also effectively predict the prognosis of disease-free survival (DFS) with an AUC of 0.749 at 1 year and 0.830 at 3 years. We further screened the genes from the tumor prediction model to establish a unique personalized panel consisting of 9 individual somatic mutation genes to detect and analyze ctDNA. It was found that ctDNA positivity decreased with the passage of time during NAC, and ctDNA status can predict NAC response and metastasis recurrence. Finally, we constructed the chemotherapy prediction model combined with the tumor prediction model and pretreatment ctDNA levels, which has a better prediction effect of pCR with the AUC value of 0.961. In this study, we established a chemotherapy predictive model with a non-invasive tool that is built based on genomic features, ctDNA status, as well as clinical characteristics for predicting pCR to recognize the responders and non-responders to NAC, and also predicting prognosis for DFS in breast cancer. Adding pretreatment ctDNA levels to a model containing gene profile mutation and clinical characteristics significantly improves stratification over the clinical variables alone.


Clinical outcomes and nadir prostate-specific antigen (PSA) according to initial PSA levels in primary androgen deprivation therapy for metastatic prostate cancer.
To investigate the clinical outcomes of metastatic prostate cancer patients and the relationship between nadir prostate-specific antigen (PSA) levels and different types of primary androgen deprivation therapy (PADT). This study utilized data from the Japan Study Group of Prostate Cancer registry, which is a large, multicenter, population-based database. A total of 2982 patients treated with PADT were enrolled. Kaplan-Meier analysis was used to compare progression-free survival (PFS) and overall survival (OS) in patients treated using combined androgen blockade (CAB) and non-CAB therapies. The relationships between nadir PSA levels and PADT type according to initial serum PSA levels were also investigated. Among the 2982 enrolled patients, 2101 (70.5 %) were treated with CAB. Although CAB-treated patients had worse clinical characteristics, their probability of PFS and OS was higher compared with those treated with a non-CAB therapy. These results were due to a survival benefit with CAB in patients with an initial PSA level of 500-1000 ng/mL. Nadir PSA levels were significantly lower in CAB patients than in non-CAB patients with comparable initial serum PSA levels. A small survival benefit for CAB in metastatic prostate cancer was demonstrated in a Japanese large-scale prospective cohort study. The clinical significance of nadir PSA levels following PADT was evident, but the predictive impact of PSA nadir on OS was different between CAB and non-CAB therapy.


Talking About Prescription Opioid Misuse: The Effect of Family Communication Patterns and Messages Advocating Direct and Indirect Communication.
Families play an important role in addressing substance misuse and addiction. Extant literature suggests patterns of communication within families influence the ways in which they engage loved ones who may be misusing substances like prescription opioids. However, little is known regarding how strategic health messages about family communication influence individuals' intentions to engage in conversations about substance misuse. Applying a normative approach, we conducted a (2 × 2) between-participants experiment examining whether messages advocating indirect (versus direct) communication are more effective for individuals (n = 613) who describe their family as having a low (versus high) conversation orientation. Univariate analysis of variance tests show match effects for message attitudes and message elaboration. For intentions to talk with a loved one about the risks of OUD, there was only evidence of a matching effect between the message advocating indirect communication with low conversation audiences. Both message types were equally effective at influencing intentions for high conversation participants. These findings suggest message designers should consider the kinds of communication behaviors and actions advocated in appeals targeting family members. Messages that are inclusive of the conversation dynamics of particular audiences may have greater effect. In particular, for low conversation audiences, messages advocating an indirect approach may be more effective at motivating intentions to engage someone who is misusing opioids.


Photosystem II reaction centre quenching: mechanisms and physiological role.
Dissipation of excess absorbed light energy in eukaryotic photoautotrophs through zeaxanthin- and DeltapH-dependent photosystem II antenna quenching is considered the major mechanism for non-photochemical quenching and photoprotection. However, there is mounting evidence of a zeaxanthin-independent pathway for dissipation of excess light energy based within the PSII reaction centre that may also play a significant role in photoprotection. We summarize recent reports which indicate that this enigma can be explained, in part, by the fact that PSII reaction centres can be reversibly interconverted from photochemical energy transducers that convert light into ATP and NADPH to efficient, non-photochemical energy quenchers that protect the photosynthetic apparatus from photodamage. In our opinion, reaction centre quenching complements photoprotection through antenna quenching, and dynamic regulation of photosystem II reaction centre represents a general response to any environmental condition that predisposes the accumulation of reduced Q(A) in the photosystem II reaction centres of prokaryotic and eukaryotic photoautotrophs. Since the evolution of reaction centres preceded the evolution of light harvesting systems, reaction centre quenching may represent the oldest photoprotective mechanism.


[Autoimmune reactions of the humoral type in viral hepatitis].
Antibodies against antigens of hepatocyte cell membranes are formed in half the patients with hepatitis A and B in one-third of those with pseudotuberculosis. They occur in the period preceding the patients' convalescence rather than in the acute stage of the disease. The recovery of hepatitis patients in whom liver antibodies formed ensued significantly more rapidly than in persons with no autoantibodies. Antibodies against antigens of hepatocyte cell membranes were detectable in different clinical disease patterns. They did not influence the disease progress and witnessed hepatocyte injuries not only in the icteric but also in scarlatiniform and and abdominal disease patterns.


Pilot Studies of VOC Exposure Profiles during Surgical Operations.
Volatile organic chemical exposure resulting from surgical operations is common in operating room personnel. The potential risk of long-term exposure to these low-level chemicals is always a concern. This study was conducted in an area hospital located in northern Taiwan to investigate the internal exposure scenario for operating room personnel. Breath samples were collected before and after surgery, whereas area samples were collected during the surgeries in process. There were 18 volatile organic compounds identified in the samples with gas chromatography-mass spectrometry. The average concentrations of sevoflurane (P = 0.0082), dimethyl sulfide (P = 0.0550), and methyl methacrylate (P = 0.0606) in breath samples collected after surgical operations were significantly higher compared to those obtained before surgical operations, whereas only slight elevations were present for benzene and hexamethyldisiloxane (P < 0.100). In addition, electrosurgical smoke-related chemicals, such as benzene, toluene, ethylbenzene, and m/p-xylene, also presented higher levels in operating room samples compared to the control area. Specifically, the findings in this preliminary study suggested the associations of elevated exposure to sevoflurane across various surgeries to methyl methacrylate with orthopedic surgery and to hexamethyldisiloxane with conventional electrosurgical units. Future study is warranted to explore the short-term high-level chemical exposure in operating rooms and to propose effective preventive measures accordingly to keep any exposure to chemicals at the lowest practical level.


Links between Schwann cells and melanocytes in development and disease.
Melanocytes are pigment-producing cells that reside in the skin, eyes, ears, heart, and central nervous system meninges of mammals. Schwann cells are glial cells, which closely associate with peripheral nerves, myelinating, and sheathing them. Melanocytes and Schwann cells both arise from the neural crest during development, and some melanocytes arise directly from Schwann cell precursors lining developing spinal nerves. In this review, we explore the connections between melanocytes and Schwann cells in development and transformation.


Radionuclide evaluation of pulmonary function following hemithorax irradiation of normal dogs with 60Co or fast neutrons.
To assess the effects of therapeutic doses of radiation on regional pulmonary function, 28 dogs had serial chest radiographs and quantitative ventilation, perfusion, and aerosol studies at three-, six-, nine-, and twelve-month intervals after hemithorax irradiation with 60-Co-gamma photons or 15-MeV fast neutrons. Findings showed marked reductions in perfusion and aerosol deposition in the irradiated lungs of dogs in the high-dose photon and neutron groups and in the medium-dose neutron group. The effects of large photon and small neutron doses were similar (neutron RBE = 4.5). Radionuclide studies were found to be useful for detecting these alterations.


Early outcomes with a single-sided access endovascular stent.
The objective of this study was to report the 1-year follow-up study results of the new Horizon stent graft (Endospan, Herzliya, Israel) from two different prospective consecutive trials. The Horizon abdominal aortic aneurysm stent graft system is a 14F profile system requiring only a single access site. It consists of three modules, introduced separately: base limb (iliac to iliac limb); distal aortic limb; and proximal aortic limb with a bare suprarenal crown and active fixation. Data from the first in man (FIM) clinical study with 10 patients enrolled and the pivotal study with 30 patients were analyzed. Outcomes measured were freedom from major adverse events (MAEs) including all-cause mortality, myocardial infarction, renal failure, respiratory failure, paraplegia, stroke, bowel ischemia, and procedural blood loss ≥1000 mL. Performance end points included successful delivery and deployment of the device, freedom from aneurysm growth ≥5 mm, type I or type III endoleak, stent graft occlusion, conversion to open surgery, rupture, and stent graft migration. In the FIM study, one conversion to open surgery with >1000 mL of blood loss was registered perioperatively. In the pivotal study, no perioperative MAE was registered. Overall, at 1-year follow-up, two deaths and one aneurysm growth unrelated to endoleak were registered. The results of both the FIM and pivotal studies demonstrated that 39 of 40 procedures were successful for delivery and deployment of the Horizon stent graft. No MAE was registered during the follow-up. The primary safety and performance end points were met in both studies.


The pathogenesis of radiation nephropathy in the dog.
Canine kidneys were biopsied at 2-week intervals from 3 to 13 weeks after a single 15-Gy dose of intraoperatively delivered 6 MeV electrons to determine the pathogenesis and dose-limiting tissue for radiation nephropathy. The data suggest that the dose and time after irradiation determine the dose-limiting tissue. The early functional survival of canine kidneys appeared to be dependent on parenchymal cell killing. Histologic changes in epithelial cells were seen as early as 3 weeks after irradiation. The parenchyma decreased to 50% of the preirradiation volume by 9 weeks but repopulated to near normal by 11 weeks. A second wave of depopulation, possibly due to perivascular fibrosis, was evident at 13 weeks. Previous investigators have demonstrated progressively extensive changes in renal vessels after irradiation. In this study, permanent vascular damage was seen at 3 weeks; however, most early changes in vessel walls proved to be temporary and probably resulted from atrophic vasoconstriction following parenchymal depopulation. Vessel dimensions returned to near normal as the parenchyma repopulated; however, a "histohematic barrier" created by progressively increasing perivascular fibrosis may cause a reduction in oxygen and nutrient support of the parenchyma and permanent loss of renal function.


DC-specific ICAM-3-grabbing nonintegrin mediates internalization of HIV-1 into human podocytes.
Human immunodeficiency virus (HIV)-1 has been demonstrated to contribute to the pathogenesis of HIV-associated nephropathy. In renal biopsy studies, podocytes have been reported to be infected by HIV-1. However, the mechanism involved in HIV-1 internalization into podocytes is not clear. In the present study, we evaluated the occurrence of HIV-1 internalization into conditionally immortalized human podocytes and the mechanism involved. Human podocytes rapidly internalized R5 and X4 HIV-1 primary strains via an endocytosis-dependent pathway, without establishing a productive infection. The HIV-1 internalization was dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) receptor mediated. The role of DC-SIGN was confirmed by using specific blocking antibodies and transfection with small interfering (si) RNA/DC-SIGN. Since podocyte HIV-1 trafficking was not altered by pH-modulating agents, it appeared that HIV-1 routing occurred through nonacid vesicular compartments. Interestingly, transfection of podocytes with neither siRNA/caveolin-1 nor siRNA/clathrin heavy chain inhibited podocyte viral accumulation. Thus it appears that clathrin-coated vesicles and caveosomes may not be contributing to HIV-1-associated membrane traffic.


Resolving fast and slow motions in the internal loop containing stem-loop 1 of HIV-1 that are modulated by Mg2+ binding: role in the kissing-duplex structural transition.
Stem loop 1 (SL1) is a highly conserved hairpin in the 5'-leader of the human immunodeficiency virus type I that forms a metastable kissing dimer that is converted during viral maturation into a stable duplex with the aid of the nucleocapsid (NC) protein. SL1 contains a highly conserved internal loop that promotes the kissing-duplex transition by a mechanism that remains poorly understood. Using NMR, we characterized internal motions induced by the internal loop in an SL1 monomer that may promote the kissing-duplex transition. This includes micro-to-millisecond secondary structural transitions that cause partial melting of three base-pairs above the internal loop making them key nucleation sites for exchanging strands and nanosecond rigid-body stem motions that can help bring strands into spatial register. We show that while Mg2+ binds to the internal loop and arrests these internal motions, it preserves and/or activates local mobility at internal loop residues G272 and G273 which are implicated in NC binding. By stabilizing SL1 without compromising the accessibility of G272 and G273 for NC binding, Mg2+ may increase the dependence of the kissing-duplex transition on NC binding thus preventing spontaneous transitions from taking place and ensuring that viral RNA and protein maturation occur in concert.


A novel high-resolution melting analysis approach for rapid detection of vancomycin-resistant enterococci.
Vancomycin-resistant enterococci (VRE) are resistant to most classes of antibiotics. Diagnosis of VRE using standard methods takes 2 to 5 days. Development of a rapid PCR-assay that detects and identifies resistant genes in bacteria would provide time-critical information on the presence of VRE in clinical samples allowing early treatment and management of infected patients. Investigate the use of high resolution melting analysis (HRMA) and 16S-rRNA-PCR approach for rapid and cost-effective identification of VRE. Descriptive antibiotic susceptibility studies. Manchester Academic Health Sciences Centre and School of Translational Medicine, University of Manchester, UK, and Department of Clinical Laboratory Sciences, Taibah University, Saudi Arabia. PCR-HRMA using 16S-rRNA V1-primers was used to detect and identify VRE. DNA from different strains of vancomycin-resistant and -sensitive Enterococcus faecalis (VSE) and Enterococcus faecium were amplified using V1-primer followed by HRMA in a single run. Differentiation of VRE from VSE was based on curve shapes generated against reference organisms (Bacteroides fragilis). Amplification curves and difference plots for VRE and VSE. Difference plots were generated for all vancomycin-resistant and -sensitive E faecalis and E faecium strains by subtracting their fluo.rescence melting profile from that of a reference-species B fragilis. A characteristic curve shape was produced by vancomycin-sensitive E faecalis and E faecium. However, vancomycin-resistant strains of these bacteria were associated with a markedly different curve shape facilitating a clear differentiation. The 16S-PCR-HRMA approach has the potential for detecting vancomycin-resistant E faecium and E faecalis. Data with VRE provide the basis for combining VRE identification with pathogens speciation in a rapid, cheap assay able to identify a pathogen as an Enterococcus and whether it is vancomycin-sensitive or -resistant E faecium or E faecalis in a single PCR and HRMA run. Tested on specific, but not all, reference Enterococcus species and clinical isolates. None.


Clonal Hematopoiesis From Next Generation Sequencing of Plasma From a Patient With Lung Adenocarcinoma: A Case Report.
Reliable and accurate next generation sequencing (NGS) technologies are important in precision medicine. Analysis using currently available NGS genomic tests is conducted on cancer-derived DNA collected from tumor tissue, blood, or both. Clonal hematopoiesis (CH) produces a detectable somatic clonal mutation that is commonly associated with clonal expansion of hematopoietic cells with age and genomic analysis of blood samples can be used to detect CH. A 74-year-old Korean male had lung adenocarcinoma with a metastasis to the left scapula. He underwent palliative radiotherapy to the left scapula and received multi-line chemotherapies. After disease progression, he underwent re-biopsy of the metastatic tumor tissue from lung cancer and concomitant blood sampling. NGS genomic testing revealed no significant genomic mutation in the tumor tissue DNA but showed the TP53 mutation C135Y in peripheral blood DNA. To investigate the discordance between the genotyping results in tumor tissue and blood, we tested for the TP53 mutation using a target sequencing test in blood and normal oral mucosa. The TP53 mutation C135Y was only detected in the blood sample, confirming the presence of TP53-mutated CH. We should be aware of different characteristics in NGS genomic testing including sample type such as tumor, blood, or paired specimens. Performing genomic testing on paired tumor and blood samples is effective for discriminating mutations derived from CH from germline mutations and somatic mutations in tumor cells.


Obesity as a risk factor for nosocomial infections in trauma patients.
Obesity, like multiple trauma, is associated with an inflammatory condition that leads to an immunodeficient state. Obese trauma patients are thus thought to be at higher risk of infection compared to patients of normal body mass. Despite this risk, studies to date have not defined obesity as an independent risk factor for infection in trauma patients. Retrospective data were collected on 1,024 patients admitted to a Level I trauma center during a 12-month period. Obesity was defined as a body mass index (BMI) >or= 30 kg/m(2). Outcomes analyzed included urinary tract infection, pneumonia, septicemia, and wound infection and Clostridium difficile infection. Multiple logistic regression was used to evaluate the contribution of each BMI category to infection while adjusting for comorbidities, age, gender, Injury Severity Score (ISS), hospital and ICU lengths of stay, and number of ventilator days. Obesity prevalence was 30.6%. Obese patients had longer hospital length of stay, with similar ISS, number of ventilator days, and ICU length of stay. The overall rate of infections was 8.7%. Variables independently associated with increased risk of infections were BMI, age, ISS, ICU length of stay, hospital length of stay, and multiple comorbidities. The risks of infections according to each BMI category were: BMI <or= 25 kg/m(2), 4.2%, BMI 25 to 29 kg/m(2), 9.5%, odds ratio (OR) 2.65 (CI 0.72 to 5.72); BMI 30 to 39 kg/m(2), 12%, OR 4.69 (CI 2.18 to 10.08); and BMI >or= 40 kg/m(2), 20.3%, OR 5.91 (CI 2.18 to 16.01). Pulmonary and wound infections were significantly more frequent in obese patients. In this retrospective study, obesity was shown to be an independent risk factor for nosocomial infection after trauma. Prospective studies would clarify the reasons associated with this increased risk of infections in obese trauma patients.


liver-enriched gene 1a and 1b encode novel secretory proteins essential for normal liver development in zebrafish.
liver-enriched gene 1 (leg1) is a liver-enriched gene in zebrafish and encodes a novel protein. Our preliminary data suggested that Leg1 is probably involved in early liver development. However, no detailed characterization of Leg1 has been reported thus far. We undertook both bioinformatic and experimental approaches to study leg1 gene structure and its role in early liver development. We found that Leg1 identifies a new conserved protein superfamily featured by the presence of domain of unknown function 781 (DUF781). There are two copies of leg1 in zebrafish, namely leg1a and leg1b. Both leg1a and leg1b are expressed in the larvae and adult liver with leg1a being the predominant form. Knockdown of Leg1a or Leg1b by their respective morpholinos specifically targeting their 5'-UTR each resulted in a small liver phenotype, demonstrating that both Leg1a and Leg1b are important for early liver development. Meanwhile, we found that injection of leg1-ATG(MO), a morpholino which can simultaneously block the translation of Leg1a and Leg1b, caused not only a small liver phenotype but hypoplastic exocrine pancreas and intestinal tube as well. Further examination of leg1-ATG(MO) morphants with early endoderm markers and early hepatic markers revealed that although depletion of total Leg1 does not alter the hepatic and pancreatic fate of the endoderm cells, it leads to cell cycle arrest that results in growth retardation of liver, exocrine pancreas and intestine. Finally, we proved that Leg1 is a secretory protein. This intrigued us to propose that Leg1 might act as a novel secreted regulator that is essential for liver and other digestive organ development in zebrafish.


Functional annotation of orthologs in metagenomes: a case study of genes for the transformation of oceanic dimethylsulfoniopropionate.
Dimethylsulfoniopropionate (DMSP) is produced mainly by phytoplankton and bacteria. It is relatively abundant and ubiquitous in the marine environment, where bacterioplankton make use of it readily as both carbon and sulfur sources. In one transformation pathway, part of the molecule becomes dimethylsulfide (DMS), which escapes into the atmosphere and plays an important role in the sulfur exchange between oceans and atmosphere. Through its other dominant catabolic pathway, bacteria are able to use it as sulfur source. During the past few years, a number of genes involved in its transformation have been characterized. Identifying genes in taxonomic groups not amenable to conventional methods of cultivation is challenging. Indeed, functional annotation of genes in environmental studies is not straightforward, considering that particular taxa are not well represented in the available sequence databases. Furthermore, many genes belong to families of paralogs with similar sequences but perhaps different functions. In this study, we develop in silico approaches to infer protein function of an environmentally important gene (dmdA) that carries out the first step in the sulfur assimilation from DMSP. The method combines a set of tools to annotate a targeted gene in genome databases and metagenome assemblies. The method will be useful to identify genes that carry out key biochemical processes in the environment.


In Silico Investigation on the Molecular Behavior and Structural Stability of the Rosette Nanotubes as the Drug Vehicles for Paclitaxel, an Anti-Cancer Drug.
Most anticancer drugs affect healthy cells in addition to cancer cells, causing severe side effects. Targeted delivery by nano-based drug delivery systems (NDDS) can reduce these severe side effects while maintaining therapeutic efficacy. This work introduced rosette nanotube (RNT) as a potential drug vehicle for paclitaxel (PTX) due to its self-assembling property, biocompatibility, amphiphilicity, and low toxicity. Molecular dynamics (MD) simulations aided with molecular mechanics Poisson Boltzmann surface area (MMPBSA) analysis are used here to investigate the molecular behavior and the loading energetics of each type of RNT (K1, xK1, and iEt-xK1) with PTX. Analysis showed that the most probable configuration of PTX is on either end of each RNT. The binding free energies (-117.74 to -69.29 kJ/mol) when PTX is closer to one end were stronger than when it is in the inner channel (-53.51 to -40.88 kJ/mol). The latter alludes to the encapsulation of the PTX by each RNT. Thus, loading is possible by encapsulation during the self-assembly process given the favorable estimated binding free energies. Based on the results, RNT has potential as a drug vehicle for PTX, which warrants further investigation.


Presenilin-interacting proteins.
Familial Alzheimer's disease (FAD) accounts for 5-10% of deaths from Alzheimer's disease (AD), and approximately 50% of these cases have been definitely linked to missense mutations in three genes, encoding the amyloid precursor protein (APP), presenilin 1 (PS1) and presenilin 2 (PS2). Of these, the vast majority of FAD-linked mutations are within PS1. There has been an extensive effort to identify proteins that functionally interact with PS1 and PS2 because of their clear roles in FAD. The goal of this review is to describe these proteins and to discuss in more detail the probable biological functions of a subset of the better-studied interacting proteins. In particular, the review examines APP, Notch, nicastrin, modifier of cellular adhesion (MOCA), beta-catenin, and the group of proteins involved in cell death, calcium metabolism and cell adhesion. We argue that, although a few of the interacting proteins are unambiguously involved in well-studied cellular pathways, their exact roles within these pathways have not been clearly defined, and indeed might vary between cell types. We also question the physiological relevance of some of the work linking PS to cell death pathways. Finally, we point out the value of using flies and worms to sort out the often contradictory work in the PS field, and we mention how knowledge of PS-interacting pathways will contribute to the development of new therapeutic strategies in AD.


Coronary artery ectasia: a ten-year experience in a tertiary hospital in Singapore.
Coronary artery ectasia (CAE) is a well-recognised, albeit rare, angiographic finding of abnormal coronary dilatation. To our best knowledge, its incidence has never been described in Singapore. We aimed to determine the incidence of CAE at our teaching hospital and to describe patient and angiographic characteristics. Cases were identified through a search of our hospital's computer database and all records were reviewed. Over the 10-year study period (1992-2001), 8641 patients underwent coronary angiography. CAE was diagnosed in 104 patients, giving an incidence of 1.2%. The majority (74%) were male. The median age was 54 years (range, 25 to 79 years). Sixty-six per cent of patients were Chinese, 19% Malays, 12% Indians and 3% other races. Concomitant diabetes mellitus was present in 31%, hypertension in 58% and dyslipidaemia in 63% of patients. Co-existent obstructive coronary artery disease was present in 82% of cases. The frequency of arterial involvement was: right coronary artery (RCA), 65%; left anterior descending artery (LAD), 48%; circumflex artery (CX), 43%; and left main artery (LM), 2%. CAE affected only 1 major vessel in 62% of cases and all 3 vessels in 20%. Eighteen patients were advised to undergo bypass surgery, while percutaneous coronary intervention was recommended in 26 patients. The incidence of ectasia was 1.2%. The majority of patients were males in their sixth decade with underlying dyslipidaemia or hypertension. CAE was associated with obstructive coronary artery disease in more than 80% of cases. The RCA was most commonly affected and most patients had single vessel involvement.


State-to-State Rates for the D + H2(v = 1, j = 1) rarr HD(v', j') + H Reaction: Predictions and Measurements.
A fully quantal wavepacket approach to reactive scattering in which the best available H(3) potential energy surface was used enabled a comparison with experimentally determined rates for the D + H(2)(v = 1, j = 1) --> HD(v' = 0, 1, 2; j') + H reaction at significantly higher total energies (1.4 to 2.25 electron volts) than previously possible. The theoretical results are obtained over a sufficient range of conditions that a detailed simulation of the experiment was possible, thus making this a definitive comparison of experiment and theory. Good to excellent agreement is found for the vibrational branching ratios and for the rotational distributions within each product vibrational level. However, the calculated rotational distributions are slightly hotter than the experimentally measured ones. This small discrepancy is more marked for products for which a larger fraction of the total energy appears in translation. The most likely explanation for this behavior is that refinements are needed in the potential energy surface.


Tityus serrulatus venom and toxins Ts1, Ts2 and Ts6 induce macrophage activation and production of immune mediators.
Scorpion envenomation induces a systemic immune response, and neurotoxins of venom act on specific ion channels, modulating neurotransmitter release or activity. However, little is known about the immunomodulatory effects of crude venom from scorpion Tityus serrulatus (TsV) or its toxins (Ts1, Ts2 and Ts6) in combination with lipopolysaccharide (LPS). To investigate the immunomodulatory effects of TsV and its toxins (Ts1, Ts2 and Ts6), J774.1 cells were stimulated with different concentrations (25, 50 and 100 μg/mL) of venom or toxins pre-stimulated or not with LPS (0.5 μg/mL). Macrophage cytotoxicity was assessed, and nitric oxide (NO) and cytokine production were analyzed utilizing the culture supernatants. TsV and its toxins did not produce cytotoxic effects. Depending on the concentrations used, TsV, Ts1 and Ts6 stimulated the production of NO, interleukin (IL)-6 and tumor necrosis factor (TNF)-α in J774.1 cells, which were enhanced under LPS co-stimulation. However, LPS + Ts2 inhibited NO, IL-6 and TNF-α production, and Ts2 alone stimulated the production of IL-10, suggesting an anti-inflammatory activity for this toxin. Our findings are important for the basic understanding of the mechanisms involved in macrophage activation following envenomation; additionally, these findings may contribute to the discovery of new therapeutic compounds to treat immune-mediated diseases.


[Nutritional support in chylothorax secondary to lymphoma].
Chylothorax is a well-known, albeit infrequent, complication in certain neoplasias including lymphomas. The continuing loss of chyle leads to a profound deterioration in patients' nutritional and immunological status, prevented only by appropriate early nutritional support. There is currently some dispute over its handling, particularly with regard to the most appropriate type of nutritional support and the suitability of conservative treatment versus surgery. The present paper describes the case of a 41-year-old patient diagnosed as having secondary chylothorax following B-cell lymphoma where mixed nutritional support (fat-free enteral nutrition by mouth and total parenteral nutrition) was given, followed by talc pleurodesis, with optimal results. There is a review of the literature and a discussion of the most controversial aspects of its management.


nab-Paclitaxel plus gemcitabine for metastatic pancreatic cancer: long-term survival from a phase III trial.
Positive findings from the phase III MPACT trial led to the regulatory approval of nab-paclitaxel plus gemcitabine as a treatment option for patients with metastatic pancreatic cancer. This report is an update of overall survival (OS) based on longer follow-up. Patients (n = 861) with metastatic pancreatic cancer and a Karnofsky performance status of 70 or greater were randomly assigned one to one to receive nab-paclitaxel + gemcitabine or gemcitabine alone. Efficacy data for this post hoc analysis were collected through May 9, 2013. Exploratory analyses of carbohydrate antigen 19-9 (CA19-9) and neutrophil-to-lymphocyte ratio (NLR) were conducted. The primary efficacy endpoint was OS, which was analyzed for all randomly assigned patients by the Kaplan-Meier method. All statistical tests were two-sided. The median OS was statistically significantly longer for nab-paclitaxel plus gemcitabine vs gemcitabine alone (8.7 vs 6.6 months, hazard ratio [HR] = 0.72, 95% confidence interval [CI] = 0.62 to 0.83, P < .001). Long-term (>three-year) survivors were identified in the nab-paclitaxel plus gemcitabine arm only (4%). In pooled treatment arm analyses, higher CA19-9 level and NLR at baseline were statistically significantly associated with worse OS. There appeared to be a treatment effect for OS favoring nab-paclitaxel plus gemcitabine over gemcitabine alone in poor-prognosis subgroups defined by these factors (HR = 0.612, P < .001 for CA19-9 level ≥ median and HR = 0.81, P = .079 for NLR > 5). These data confirm and extend the primary report of OS, supporting the superior efficacy of nab-paclitaxel plus gemcitabine over gemcitabine alone. Subgroup analyses support the relevance of CA 19-9 and NLR as prognostic markers in metastatic pancreatic cancer.


Targeting uracil-DNA glycosylases for therapeutic outcomes using insights from virus evolution.
Ung-type uracil-DNA glycosylases are frontline defenders of DNA sequence fidelity in bacteria, plants and animals; Ungs also directly assist both innate and humoral immunity. Critically important in viral pathogenesis, whether acting for or against viral DNA persistence, Ungs also have therapeutic relevance to cancer, microbial and parasitic diseases. Ung catalytic specificity is uniquely conserved, yet selective antiviral drugging of the Ung catalytic pocket is tractable. However, more promising precision therapy approaches present themselves via insights from viral strategies, including sequestration or adaptation of Ung for noncanonical roles. A universal Ung inhibition mechanism, converged upon by unrelated viruses, could also inform design of compounds to inhibit specific distinct Ungs. Extrapolating current developments, the character of such novel chemical entities is proposed.


Large Nonlinear Transverse Conductivity and Berry Curvature in KTaO3 Based Two-Dimensional Electron Gas.
Two-dimensional electron gas (2DEG) at oxide interfaces exhibits various exotic properties stemming from interfacial inversion and symmetry breaking. In this work, we report large nonlinear transverse conductivities in the LaAlO3/KTaO3 interface 2DEG under zero magnetic field. Skew scattering was identified as the dominant origin based on the cubic scaling of nonlinear transverse conductivity with linear longitudinal conductivity and 3-fold symmetry. Moreover, gate-tunable nonlinear transport with pronounced peak and dip was observed and reproduced by our theoretical calculation. These results indicate the presence of Berry curvature hotspots and thus a large Berry curvature triplet at the oxide interface. Our theoretical calculations confirm the existence of large Berry curvatures from the avoided crossing of multiple 5d-orbit bands, orders of magnitude larger than that in transition-metal dichalcogenides. Nonlinear transport offers a new pathway to probe the Berry curvature at oxide interfaces and facilitates new applications in oxide nonlinear electronics.


HIV-1 Entry and Prospects for Protecting against Infection.
Human Immunodeficiency Virus type-1 (HIV-1) establishes a latent viral reservoir soon after infection, which poses a major challenge for drug treatment and curative strategies. Many efforts are therefore focused on blocking infection. To this end, both viral and host factors relevant to the onset of infection need to be considered. Given that HIV-1 is most often transmitted mucosally, strategies designed to protect against infection need to be effective at mucosal portals of entry. These strategies need to contend also with cell-free and cell-associated transmitted/founder (T/F) virus forms; both can initiate and establish infection. This review will discuss how insight from the current model of HIV-1 mucosal transmission and cell entry has highlighted challenges in developing effective strategies to prevent infection. First, we examine key viral and host factors that play a role in transmission and infection. We then discuss preventive strategies based on antibody-mediated protection, with emphasis on targeting T/F viruses and mucosal immunity. Lastly, we review treatment strategies targeting viral entry, with focus on the most clinically advanced entry inhibitors.


Office gynecologic procedures.
The screening and detection of endometrial carcinoma can be done safely and effectively in the outpatient setting using recently developed endometrial biopsy devices. The indications and techniques of endometrial biopsy are described in this article. In addition, the use of cryosurgery of the uterine cervix for the treatment of benign cervical disease as well as cervical intraepithelial neoplasia is reviewed.


The Value of Clinical Colorectal Cancer Registries in Colorectal Cancer Research: A Systematic Review.
Clinical colorectal cancer registries (CCCRs) are potentially powerful tools in colorectal cancer research. They are resource intensive, but to our knowledge, no formal review of their value exists. While quality control, clinical audit, and benchmarking are important factors in assessing the value of maintaining CCCRs, they are difficult to quantify. This study focuses on registry research output as a measure of value; the study hypothesizes that CCCRs do not produce sufficient published research output of clinical significance to justify the resources required to maintain them. To assess the value of maintaining CCCRs by identifying and characterizing existing CCCRs and measuring their comparative research impact. We searched MEDLINE (PubMed) and Google Scholar for articles published from January 1990 to July 2016 that identified multi-institutional CCCRs with peer-reviewed published outcomes. Purely population-based registries were excluded. We then searched the same databases in the same time period for articles that were published by each included CCCR. The articles must have been based on outcomes relating to individual CCCR data. We categorized published outcomes into oncological, surgical, or other outcomes. We measured the research impact of each CCCR using the number of articles, citation index, impact factor, and Altmetric score. A total of 18 CCCRs were identified, with sample sizes between 104 and 1 400 000 cases. Data fields, published aims, and outcomes were similar between registries. The most frequently published outcomes related to anastomotic leak following colorectal surgery. The National Cancer Database formed the basis of the highest number of publications (66), the Northern Region Colorectal Cancer Audit Group had the highest median article citation number (28.5), the National Bowel Cancer Audit had the highest median impact factor (4.72), and the National Cancer Database had the highest median Altmetric score (4.5). There is a significant body of colorectal cancer outcomes research generated from the CCCRs. However, given the enormous resources required, the overall research output and impact of CCCRs is low in proportion to the size of the data sets. These registries hold key oncological and surgical outcomes data; focusing on data linkage between registries and developing automated data collection will enable international comparisons in colorectal cancer management and will increase the research impact of CCCRs, thereby increasing their value.


Antioxidant role of L-carnitine in an experimental model of oxidative stress induced by increased fructose consumption.
To evaluate the role of L-carnitine (LC) on fructose-induced oxidative stress in Holtzman rats. An experimental study was carried out during 56 days, in patients assigned to 4 groups: control, control+LC, fructose and fructose+LC. Patients in the fructose group received treatment during 56 days, and those in the LC groups were treated during the last 28 days. Fructose was given on demand and LC was administered orally at a dose of 500 g/kg/24 h. Lipid peroxidation (MDA), superoxide dismutase activity, free LC and mitochondrial and post-mitochondrial proteins were measured in liver tissue. Glycemia, insulin and the homeostasis model assessment of insulin resistance (HOMA-IR) were measured in blood plasma. We measured insulin concentration and studied the histology of pancreatic tissue. LC treatment showed a decrease (p < 0.05) of MDA when compared to the control group (21.73 ± 5.36 nmol/g tissue vs. 64.46 ± 7.87 nmol/g tissue). Mitochondrial and post-mitochondrial proteins increased (p < 0.05) in comparison to the control group; pancreatic insulin also increased when compared to the control (341.8 ± 42.3 μUI/ml vs. 70.1 ± 9.6 μUI/ml, p<0.05). The role of LC against fructose-induced oxidative stress did not show any decrease of MDA, but decreased (p < 0.05) SOD Cu/Zn activity (9.39 ± 1.5 USOD/mg protein vs. 13.52 ± 1.5 USOD/mg protein). We observed that LC improves HOMA-IR in blood plasma. Histological analysis of the pancreas showed that the presence of LC increased the number and size of the islets of Langerhans. LC favors changes in the oxidative metabolism and it also contributes to glycemic homeostasis when fructose is consumed.


Quantitative requirement for ATP for active transport in isolated renal cells.
We investigated the quantitative relationship between cellular ATP concentration and Na+-K+-ATPase activity as measured by ouabain-sensitive 86Rb influx in rabbit proximal renal cells. Cellular ATP was reduced in a stepwise manner by rotenone (10(-7) to 10(-5) M) and was increased by 10 mM adenosine. During these maneuvers, ouabain-sensitive 86Rb influx was linearly related to cellular ATP and did not saturate up to 9.9 mM ATP. In contrast, Na+-K+-ATPase activity in membranes prepared from these cells saturated at 2.0 mM ATP at various sodium (10-100 mM) and potassium (4-100 mM) concentrations. Sodium-dependent phosphate uptake and alpha-methylglucoside (alpha-MG) uptake were both inhibited to a similar degree when cellular ATP was reduced. We conclude that 1) the ATP requirement for saturation of Na+-K+-ATPase is higher in intact renal cells than in the membranes, and 2) the uptake of phosphate and alpha-MG are similarly influenced by reduction in ATP. This effect of ATP on phosphate and AMG uptake is most likely an indirect one and is secondary to changes in the sodium gradient across the cell.


Design of multichannel DWDM fiber Bragg grating filters by Lagrange multiplier constrained optimization.
We present the synthesis of multi-channel fiber Bragg grating (MCFBG) filters for dense wavelength-division-multiplexing (DWDM) application by using a simple optimization approach based on a Lagrange multiplier optimization (LMO) method. We demonstrate for the first time that the LMO method can be used to constrain various parameters of the designed MCFBG filters for practical application demands and fabrication requirements. The designed filters have a number of merits, i.e., flat-top and low dispersion spectral response as well as single stage. Above all, the maximum amplitude of the index modulation profiles of the designed MCFBGs can be substantially reduced under the applied constrained condition. The simulation results demonstrate that the LMO algorithm can provide a potential alternative for complex fiber grating filter design problems.


Clavin, a type-1 ribosome-inactivating protein from Aspergillus clavatus IFO 8605. cDNA isolation, heterologous expression, biochemical and biological characterization of the recombinant protein.
We describe the cloning and expression of a new cDNA from the filamentous fungus Aspergillus clavatus IFO 8605. This cDNA contains an open reading frame (ORF) that predicts a putative ribonuclease precursor with high similarity to the alpha-sarcin family of ribosome-inactivating proteins (RIPs). The cDNA encoding the mature protein was expressed in Escherichia coli, and the recombinant protein, a 17-kDa polypeptide designated clavin was purified and characterized. Clavin shows typical type-1 RIP properties: specific cleavage of ribosomal and synthetic RNA and inhibition of protein synthesis in cell-free and cellular systems. When selectively targeted to a tumour cell antigen by coupling to a monoclonal antibody (mAb) clavin was able to inhibit protein synthesis at nanomolar concentration. Pharmacokinetics analysis in mice indicated an elimination half-life (t1/2 beta) of 7.4 h with no particular accumulation in major organs. Liver toxicity was very limited and transient while no alteration of kidney function was observed. Clavin induced a late and very low antibody response in mice. The in vitro and in vivo biological characteristics of clavin, together with its availability in large amounts, suggest the usefulness of this toxin in the production of toxic chemical conjugates.


Combined femoral vein transposition and iliac vein to suprarenal vena cava bypass as a last resort dialysis access.
Patients undergoing hemodialysis are known to develop central venous occlusion and exhaust all options for vascular access to upper extremity sites; therefore, creating and maintaining vascular access is paramount in such patients. The present case report describes the condition of a 34-year-old woman with failed upper extremity access, frequent catheter-related issues, and multiple central venous occlusions. As a last resort, access to the lower extremity was pursued as follows: an inferior vena cava bypass was combined with a right femoral transposition fistula and a distal revascularization interval ligation procedure. This complex procedure that was carried out for the purpose of vascular access is a unique, albeit aggressive, surgical solution that resulted in autologous vascular access with a 6-month patency and also served to improve the quality of life in the seemingly hopeless case.


Phagocytized intracellular microsporidian blocks phagosome acidification and phagosome-lysosome fusion.
This study examines the relationship between phagosome acidification and phagosome-lysosome fusion events using phagocytized Glugea hertwigi spores. The incidence of lysosome fusion with Glugea spores in phagosomes of mouse peritoneal macrophages and of Tetrahymena was monitored using colloidal gold and acridine orange as labels for secondary lysosomes. Over 80% of the Glugea phagosomes remained segregated from the labeled compartments in macrophages after 60 min; this inhibition of fusion was still evident after 4 h. In Tetrahymena, Glugea spores also showed a high capacity to block fusion with secondary lysosomes (67%); however, spores coated with cationized ferritin showed an 80% fusion rate with labeled acidic compartments (i.e. lysosomes) after 60 min with both Tetrahymena and macrophages. The pH of phagosome compartments was monitored by measuring the emissions of fluorescein isothiocyanate (FITC)-labeled Glugea ingested by Tetrahymena. Tetrahymena phagosomes with FITC-Glugea did not acidify within the first hour after phagocytosis; however, phagosomes with cationized ferritin-labeled Glugea underwent acidification during this time period. This acidification took place although the capability of the host cells' lysosomes to fuse was blocked by pretreatment with poly-D-glutamic acid. The cationized ferritin bound to Glugea spores was uncoupled from the spore wall prior to fusion with colloidal gold-labeled compartments. In vitro testing showed that ferritin dissociation requires an acid pH, indicating that phagosomes acidify prior to lysosome fusion.


Herpes simplex virus type 1-induced interferon production and activation of natural killer cells in mice.
Cultures of peritoneal exudate cells (PEC) of non-immune C57BL/6 mice reacted to infectious herpes simplex virus type 1 (HSV), but not to non-infectious virions by producing interferon. Similarly, when the virus preparations were injected intraperitoneally (i.p.) and interferon was determined in the wash-out fluid, interferon could only be detected after injection of infectious HSV. All interferons were mixtures of interferon-alpha and -beta. The cell that produced interferon was not sensitive to anti-theta serum or anti-asialo-GM1 serum, it was retained on nylon wool columns and removed by plastic adherence. Silica treatment of PEC abolished HSV-induced interferon production whereas irradiation by 2000 R was without effect. Pure cultures of mouse T cells did not produce interferon when treated with infectious HSV whereas pure cultures of mouse bone marrow macrophages did produce interferon. Thus, we suggest that in PEC the cells which produced interferon were of the macrophage lineage. In further experiments we have analysed in vivo activation of natural killer (NK) cells in the peritoneal cavity of mice. Non-infectious virions were as effective as infectious HSV and doses of HSV too low to induce measurable titres of interferon caused a marked activation of NK cells.


Atraumatic osteonecrosis of the adult elbow.
Osteonecrosis is a disease that leads to joint destruction and often involves large joints, such as the hips, knees, and shoulders. Nontraumatic osteonecrosis of the adult elbow, to the best of the authors' knowledge, has not been reported. Nine adult patients with atraumatic osteonecrosis of 11 elbows were identified. The mean age at presentation was 36 years (range, 26-63 years); five patients were women and four were men. Six elbows involved the capitellum, three involved the lateral epicondyles, one involved the trochlea and radial head, and one involved medial and lateral epicondylar disease. All patients were receiving corticosteroid therapy, and no relationship between the duration or the amount of corticosteroid use and the severity of the osteonecrosis was found. Seven patients with radiographic Stage I and Stage II disease responded well to nonoperative treatments consisting of activity modification, analgesics, and a brief period of immobilization. Nonoperative treatment failed in two patients with Stage III disease, and they had core decompressions for pain relief. One patient with late Stage III disease in both elbows underwent bilateral total elbow arthroplasties. In contrast to the pediatric population, osteonecrosis of the adult elbow potentially can lead to end stage arthritis. If the osteonecrosis is diagnosed early, nonoperative treatment may be effective in relieving pain, although the long-term results of these treatments remain unknown.


Congenital mesonephric defects in male infants with mucoviscidosis.
Reports that men with mucoviscidosis were sterile and that portions of their genital tracts were atretic prompted us to investigate the genital tracts of 15 male infants with mucoviscidosis who died within the first year of life and came to necropsy. The genital tracts of all of these infants were abnormal, the abnormalities being confined to mesonephric derivatives. Hypoplastic or absent ducti efferentia, ducti epididymides, or ducti deferentia were found in all 28 specimens of epididymides, and the ducti deferentia were missing from 25 of 27 examples of spermatic cord. The seminal vesicles and the ejaculatory ducts were less frequently hypoplastic or absent. Because these abnormalities of mesonephric derivatives were present so early in life and inflammatory and obstructive changes were absent we believe that they resulted from a failure of development.


PINNet: a deep neural network with pathway prior knowledge for Alzheimer's disease.
Identification of Alzheimer's Disease (AD)-related transcriptomic signatures from blood is important for early diagnosis of the disease. Deep learning techniques are potent classifiers for AD diagnosis, but most have been unable to identify biomarkers because of their lack of interpretability. To address these challenges, we propose a pathway information-based neural network (PINNet) to predict AD patients and analyze blood and brain transcriptomic signatures using an interpretable deep learning model. PINNet is a deep neural network (DNN) model with pathway prior knowledge from either the Gene Ontology or Kyoto Encyclopedia of Genes and Genomes databases. Then, a backpropagation-based model interpretation method was applied to reveal essential pathways and genes for predicting AD. The performance of PINNet was compared with a DNN model without a pathway. Performances of PINNet outperformed or were similar to those of DNN without a pathway using blood and brain gene expressions, respectively. Moreover, PINNet considers more AD-related genes as essential features than DNN without a pathway in the learning process. Pathway analysis of protein-protein interaction modules of highly contributed genes showed that AD-related genes in blood were enriched with cell migration, PI3K-Akt, MAPK signaling, and apoptosis in blood. The pathways enriched in the brain module included cell migration, PI3K-Akt, MAPK signaling, apoptosis, protein ubiquitination, and t-cell activation. By integrating prior knowledge about pathways, PINNet can reveal essential pathways related to AD. The source codes are available at https://github.com/DMCB-GIST/PINNet.


A methodological and operative framework for the evaluation of an e-health project.
Assessing public sector ICT investments represents the premise for successful implementation of an e-health strategy. The recent literature stresses the importance of going beyond the mere financial and/or technical dimensions of the analysis. Consequently, the paper proposes an example of e-health project evaluation aiming to develop measures which get close to the notion of benefits to the different stakeholders involved: top management, patients, local community. The case study refers to an Italian health care organization that implemented a project of digitalization of its clinical reports production few years ago. Based on on-field research, different approaches are used to assess costs and benefits from different stakeholders' perspectives. The results of a multidimensional evaluation are reported to emphasize the need for different measures to assess the sustainability of an e-health project according to the financial convenience, the social role of the organization, and the contingent situation.


Single-molecule analysis reveals human UV-damaged DNA-binding protein (UV-DDB) dimerizes on DNA via multiple kinetic intermediates.
How human DNA repair proteins survey the genome for UV-induced photoproducts remains a poorly understood aspect of the initial damage recognition step in nucleotide excision repair (NER). To understand this process, we performed single-molecule experiments, which revealed that the human UV-damaged DNA-binding protein (UV-DDB) performs a 3D search mechanism and displays a remarkable heterogeneity in the kinetics of damage recognition. Our results indicate that UV-DDB examines sites on DNA in discrete steps before forming long-lived, nonmotile UV-DDB dimers (DDB1-DDB2)2 at sites of damage. Analysis of the rates of dissociation for the transient binding molecules on both undamaged and damaged DNA show multiple dwell times over three orders of magnitude: 0.3-0.8, 8.1, and 113-126 s. These intermediate states are believed to represent discrete UV-DDB conformers on the trajectory to stable damage detection. DNA damage promoted the formation of highly stable dimers lasting for at least 15 min. The xeroderma pigmentosum group E (XP-E) causing K244E mutant of DDB2 found in patient XP82TO, supported UV-DDB dimerization but was found to slide on DNA and failed to stably engage lesions. These findings provide molecular insight into the loss of damage discrimination observed in this XP-E patient. This study proposes that UV-DDB recognizes lesions via multiple kinetic intermediates, through a conformational proofreading mechanism.


Signaling Pathways Involved in the Development of Bronchopulmonary Dysplasia and Pulmonary Hypertension.
The alveolar and vascular developmental arrest in the premature infants poses a major problem in the management of these infants. Although, with the current management, the survival rate has improved in these infants, but bronchopulmonary dysplasia (BPD) is a serious complication associated with a high mortality rate. During the neonatal developmental period, these infants are vulnerable to stress. Hypoxia, hyperoxia, and ventilation injury lead to oxidative and inflammatory stress, which induce further damage in the lung alveoli and vasculature. Development of pulmonary hypertension (PH) in infants with BPD worsens the prognosis. Despite considerable progress in the management of premature infants, therapy to prevent BPD is not yet available. Animal experiments have shown deregulation of multiple signaling factors such as transforming growth factorβ (TGFβ), connective tissue growth factor (CTGF), fibroblast growth factor 10 (FGF10), vascular endothelial growth factor (VEGF), caveolin-1, wingless & Int-1 (WNT)/β-catenin, and elastin in the pathogenesis of BPD. This article reviews the signaling pathways entailed in the pathogenesis of BPD associated with PH and the possible management.


[Biocenosis of the natural foci of sapronotic infections (the results of 15-year observations)].
The summarized results of original studies, carried out in 1988-2003 by scientists of the Gamaleya Research Institute of Epidemiology and Microbiology, are discussed. The part dealing with the regularity and mechanisms of the circulation of pathogenic bacteria in the biocenosis of soil and water reservoirs includes the following subjects: population and intracellular interactions with protozoa; the evaluation of different representatives of hydrobios as hosts and the transmission along trophic biocenotic chains; the effect of algae and their metabolic products on bacterial populations; the possibility of the colonization of higher plants from the soil and the subsequent infection of rodents; the variability and clonal structure of bacterial population in its interaction with protozoa and plants; the chain of Yersinia circulation in agrocenosis. The part dealing with the mechanisms of the prolonged reservation of causative agents includes such subjects as the reversible transition of bacteria into the latent (uncultivable) state in soils and water reservoirs, as well as the biological inductors of this process; the prolonged preservation of latent bacteria in the cysts of protozoa and blue-green algae; the indication of causative agents in the natural foci of plague and pseudotuberculosis. On the basis of the original investigations and the data of literature new theoretical review in the field of the natural foci and the epidemiology of sapronotic infections have been formulated.


Ambient air pollution exposure and risk and progression of interstitial lung abnormalities: the Framingham Heart Study.
Ambient air pollution accelerates lung function decline among adults, however, there are limited data about its role in the development and progression of early stages of interstitial lung disease. To evaluate associations of long-term exposure to traffic and ambient pollutants with odds of interstitial lung abnormalities (ILA) and progression of ILA on repeated imaging. We ascertained ILA on chest CT obtained from 2618 Framingham participants from 2008 to 2011. Among 1846 participants who also completed a cardiac CT from 2002 to 2005, we determined interval ILA progression. We assigned distance from home address to major roadway, and the 5-year average of fine particulate matter (PM2.5), elemental carbon (EC, a traffic-related PM2.5 constituent) and ozone using spatio-temporal prediction models. Logistic regression models were adjusted for age, sex, body mass index, smoking status, packyears of smoking, household tobacco exposure, neighbourhood household value, primary occupation, cohort and date. Among 2618 participants with a chest CT, 176 (6.7%) had ILA, 1361 (52.0%) had no ILA, and the remainder were indeterminate. Among 1846 with a preceding cardiac CT, 118 (6.4%) had ILA with interval progression. In adjusted logistic regression models, an IQR difference in 5-year EC exposure of 0.14 µg/m3 was associated with a 1.27 (95% CI 1.04 to 1.55) times greater odds of ILA, and a 1.33 (95% CI 1.00 to 1.76) times greater odds of ILA progression. PM2.5 and O3 were not associated with ILA or ILA progression. Exposure to EC may increase risk of progressive ILA, however, associations with other measures of ambient pollution were inconclusive.


An N-acetyllactosamine-specific lectin, PFA, isolated from a moth (Phalera flavescens), structurally resembles an invertebrate-type lysozyme.
PFA (Phalera flavescens agglutinin) lectin purified from larvae of the lobster moth (P. flavescens) shows a strong binding ability specific to the N-acetyllactosamine (Galβ1-4GlcNAc) site. We determined the genomic and cDNA sequences of the PFA gene, which consists of five exons and spans approximately 5 kb of a genomic region. Surprisingly, the amino acid sequence (149 amino acids) was similar to invertebrate-type lysozymes and related proteins. The predicted tertiary structure of the PFA protein was similar to the lysozymes of clams such as the common orient clam (Meretrix lusoria) and Japanese littleneck (Venerupis philippinarum (Tapes japonica)). The PFA, however, lacks a catalytically essential amino acid, an Asp (D), which is one of the two important amino acids (Glu (E) and D) express the function of lysozyme. As a result, lysozyme activity assays indicated that PFA does not have lysozyme activity. Results suggest that the PFA gene evolved from a lysozyme gene through the loss of lysozyme activity sites and the acquisition of lectin activity during evolution of the genus Phalera.


Transcriptomic biomarkers for the accurate diagnosis of myocarditis.
Lymphocytic myocarditis is a clinically important condition that is difficult to diagnose and distinguish. We hypothesized that the transcriptome obtained from an endomyocardial biopsy would yield clinically relevant and accurate molecular signatures. Microarray analysis was performed on samples from patients with histologically proven lymphocytic myocarditis (n=16) and idiopathic dilated cardiomyopathy (n=32) to develop accurate diagnostic transcriptome-based biomarkers using multiple classification algorithms. We identified 9878 differentially expressed genes in lymphocytic myocarditis versus idiopathic dilated cardiomyopathy (fold change >1.2; false discovery rate <5%) from which a transcriptome-based biomarker containing 62 genes was identified that distinguished myocarditis with 100% sensitivity (95% confidence interval, 46 to 100) and 100% specificity (95% confidence interval, 66 to 100) and was generalizable to a broad range of secondary cardiomyopathies associated with inflammation (n=27), ischemic cardiomyopathy (n=8), and the normal heart (n=11). Multiple classification algorithms and quantitative real-time reverse-transcription polymerase chain reaction analysis further reduced this subset to a highly robust molecular signature of 13 genes, which still performed with 100% accuracy. Together, these findings demonstrate that transcriptomic biomarkers from a single endomyocardial biopsy can improve the clinical detection of patients with inflammatory diseases of the heart. This approach advances the clinical management and treatment of cardiac disorders with highly variable outcome.


Effect of Different Collection Times of Dermal Fibroblast Conditioned Medium (DFCM) on In Vitro Re-Epithelialisation Process.
A key event in wound healing is re-epithelialisation, which is mainly regulated via paracrine signalling of cytokines, chemokines, and growth factors secreted by fibroblasts. Fibroblast-secreted factors can be collected from the used culture medium, known as dermal fibroblast conditioned medium (DFCM). The goal of this study was to optimise the culture condition to acquire DFCM and evaluate its effect on keratinocyte attachment, proliferation, migration, and differentiation. Confluent fibroblasts were cultured with serum-free keratinocyte-specific (DFCM-KM) and fibroblast-specific (DFCM-FM) medium at different incubation times (Days 1, 2, and 3). DFCM collected after 3 days of incubation (DFCM-KM-3 and DFCM-FM-3) contained a higher protein concentration compared to other days. Supplementation of DFCM-KM-3 enhanced keratinocyte attachment, while DFCM-FM-3 significantly increased the keratinocyte wound-healing rate, with an increment of keratinocyte area and collective cell migration, which was distinctly different from DFCM-KM-3 or control medium. Further analysis confirmed that the presence of calcium at higher concentrations in DFCM-FM facilitated the changes. The confluent dermal fibroblasts after 3 days of incubation with serum-free culture medium produced higher proteins in DFCM, resulting in enhanced in vitro re-epithelialisation. These results suggest that the delivery of DFCM could be a potential treatment strategy for wound healing.


Retention and degradation of N-glycoproteins in the rough endoplasmic reticulum.
Recent studies have shown that newly synthesized proteins and glycoproteins are submitted to a quality control mechanism in the rough endoplasmic reticulum (ER). In this report we present two models: One model will illustrate a transient retention in rough ER leading to a further degradation of glycoproteins in the cytosol, (soluble alkaline phosphatase expressed in Man-P-Dol deficient CHO cells lines). The second model will illustrate a strict retention of glycoproteins in rough ER without degradation nor recycling through the Golgi (E1, E2 glycoproteins of Hepatitis C virus in stably transfected UHCV-11.4 cells and in infected Hep G2 cells). In both cases, oligomannoside structures are markers of these phenomena, either as free soluble released oligomannosides in the case of degradation, or as N-linked oligomannosides for strict retention in rough ER.


Rapid detection and differentiation of wild-type and three attenuated lapinized vaccine strains of classical swine fever virus by reverse transcription polymerase chain reaction.
A simple one-step reverse transcription polymerase chain reaction (RT-PCR) method was developed based on T-rich insertions in the viral genome for simultaneous detection and differentiation of wild type and vaccine strains of Classical swine fever virus (CSFV). The CSFV-specific primers were designed to contain the sequences of the T-rich insertion sites that exist uniquely in the 3' nontranslated regions (3' NTR) of the genome of lapinized CSFV vaccine strains. By using a one-step RT-PCR or a nested PCR followed by an agarose gel electrophoresis or a multicapillary electrophoresis, the wild-type and lapinized vaccine strains of CSFV in clinical samples could be detected and accurately distinguished. These assays can be applied to at least 3 attenuated lapinized vaccine strains, lapinized Philippines Coronel (LPC), hog cholera lapinized virus (HCLV), and Chinese strain (C strain). The detection limit of the wild-type virus was 6.3 TCID(50) (50% tissue culture infective dose)/ml for RT-PCR and 0.63 TCID(50)/ml for nested PCR. In previous studies, notable T-rich insertions of 12-13 nucleotides (nt) were found in the 3' NTR of the genome of lapinized vaccine strains of CSFV. However, this study discovered that 2 T-rich insertions, 42 and 36 nt in length, are present in the viral genome of lapinized vaccine strains LPC/PRK (primary rabbit kidney) and LPC/TS (Tam-Sui), respectively. These T-rich insertions of 12, 36, and 42 nt length increases the size of PCR fragments, which are favorable genetic markers for rapid detection of and differentiation between wild-type and different lapinized vaccine strains of CSFV.


Mucolipidosis II and III. The genetic relationships between two disorders of lysosomal enzyme biosynthesis.
The genetic relationships between the multiple variants of mucolipidosis II (I-cell disease) and mucolipidosis III (pseudo-Hurler polydystrophy) were investigated with a sensitive genetic complementation analysis procedure. These clinically distinct disorders have defects in the synthesis of a recognition marker necessary for the intracellular transport of acid hydrolases into lysosomes. Both disorders are associated with an inherited deficiency of a uridine diphosphate-N-acetyl-glucosamine: lysosomal enzyme precursor N-acetyl-glucosamine-phosphate transferase activity. We had previously shown that both disorders are genetically heterogeneous. Complementation analysis between mucolipidosis II and III fibroblasts indicated an identity of mucolipidosis II with one of the three mucolipidosis III complementation groups (ML IIIA), suggesting a close genetic relationship between these groups. The presence of several instances of complementation within this group suggested an intragenic complementation mechanism. Genetic complementation in heterokaryons resulted in increases in N-acetyl-glucosamine-phosphate transferase activity, as well as in the correction of lysosomal enzyme transport. This resulted in increases in the intracellular levels of several lysosomal enzymes and in the correction of the abnormal electrophoretic mobility pattern of intracellular beta-hexosaminidase. The findings demonstrate that a high degree of genetic heterogeneity exists within these disorders. N-acetyl-glucosamine-phosphate transferase is apparently a multicomponent enzyme with a key role in the biosynthesis and targeting of lysosomal enzymes.


Review: effects of anesthetics on brain circuit formation.
The results of several retrospective clinical studies suggest that exposure to anesthetic agents early in life is correlated with subsequent learning and behavioral disorders. Although ongoing prospective clinical trials may help to clarify this association, they remain confounded by numerous factors. Thus, some of the most compelling data supporting the hypothesis that a relatively short anesthetic exposure can lead to a long-lasting change in brain function are derived from animal models. The mechanism by which such changes could occur remains incompletely understood. Early studies identified anesthetic-induced neuronal apoptosis as a possible mechanism of injury, and more recent work suggests that anesthetics may interfere with several critical processes in brain development. The function of the mature brain requires the presence of circuits, established during development, which perform the computations underlying learning and cognition. In this review, we examine the mechanisms by which anesthetics could disrupt brain circuit formation, including effects on neuronal survival and neurogenesis, neurite growth and guidance, formation of synapses, and function of supporting cells. There is evidence that anesthetics can disrupt aspects of all of these processes, and further research is required to elucidate which are most relevant to pediatric anesthetic neurotoxicity.


Errors in Diagnosis of Spinal Epidural Abscesses in the Era of Electronic Health Records.
With this study, we set out to identify missed opportunities in diagnosis of spinal epidural abscesses to outline areas for process improvement. Using a large national clinical data repository, we identified all patients with a new diagnosis of spinal epidural abscess in the Department of Veterans Affairs (VA) during 2013. Two physicians independently conducted retrospective chart reviews on 250 randomly selected patients and evaluated their records for red flags (eg, unexplained weight loss, neurological deficits, and fever) 90 days prior to diagnosis. Diagnostic errors were defined as missed opportunities to evaluate red flags in a timely or appropriate manner. Reviewers gathered information about process breakdowns related to patient factors, the patient-provider encounter, test performance and interpretation, test follow-up and tracking, and the referral process. Reviewers also determined harm and time lag between red flags and definitive diagnoses. Of 250 patients, 119 had a new diagnosis of spinal epidural abscess, 66 (55.5%) of which experienced diagnostic error. Median time to diagnosis in error cases was 12 days, compared with 4 days in cases without error (P <.01). Red flags that were frequently not evaluated in error cases included unexplained fever (n = 57; 86.4%), focal neurological deficits with progressive or disabling symptoms (n = 54; 81.8%), and active infection (n = 54; 81.8%). Most errors involved breakdowns during the patient-provider encounter (n = 60; 90.1%), including failures in information gathering/integration, and were associated with temporary harm (n = 43; 65.2%). Despite wide availability of clinical data, errors in diagnosis of spinal epidural abscesses are common and involve inadequate history, physical examination, and test ordering. Solutions should include renewed attention to basic clinical skills.


Catalytic synthesis of hydroxymethyl-2-oxazolidinones from glycerol or glycerol carbonate and urea.
Oxazolidinones have been synthesized by reacting glycerol carbonate or glycerol with urea in the presence of γ-Zr phosphate as a catalyst. The conversion yield of the polyol or its carbonate depends on the temperature. Below 408 K the selectivity is 100 % with a conversion of up to 25 %, whereas increasing the temperature means that conversion yield grows, but the selectivity decreases, which makes the separation process more difficult. Starting from glycerol carbonate, two isomers, 6 and 6', are formed with a quasi 1:1 molar ratio because urea can attack the carbonate moiety on both sides of the carboxylic CO moiety. From glycerol the formation of the 6' isomer is preferred: the ratio of 6'/6 is close to 7. The oxazolidinones formed act as templates because they interact through hydrogen bonding with glycerol. The intensity of the interaction depends on the 6 or 6' isomer: DFT calculations showed that the energy was 22.6 kcal mol(-1) for 6-oxazolidinone and 25.7 kcal mol(-1) for 6'-oxazolidinone.


Methanocella arvoryzae sp. nov., a hydrogenotrophic methanogen isolated from rice field soil.
A novel hydrogenotrophic methanogen, designated strain MRE50(T), was isolated from a methanogenic consortium, which was originally established from an Italian rice field soil. Cells were non-motile rods, 1.3-2.8 μm long and 0.4-0.7 μm wide. Coccoid cells were also observed in cultures at the late-exponential phase of growth. Strain MRE50(T) grew at 37-55 °C (optimally at 45 °C), at pH 6-7.8 (optimally at pH 7.0) and in the presence of 0-20 g NaCl l(-1). The isolate utilized H(2)/CO(2) and formate for growth and methane production. Phylogenetic analyses of the 16S rRNA gene and the methanogen-specific marker gene mcrA showed that strain MRE50(T) is affiliated with the order Methanocellales, previously known as uncultured archaeal group Rice Cluster I. Based on both 16S rRNA gene and mcrA gene sequences, strain MRE50(T) was related most closely to Methanocella paludicola SANAE(T). Levels of sequence similarity were 92.5 and 86.1 %, respectively, indicating that strains MRE50(T) and Methanocella paludicola SANAE(T) represent different species within the genus Methanocella. In addition, although these strains shared phenotypic properties including cell morphology and substrate utilization, they differed with respect to susceptibility to antibiotics, and temperature and NaCl ranges for growth. Given the phenotypic differences and the distinct phylogenetic placement of the new isolate relative to the type species of the genus Methanocella, strain MRE50(T) is considered to represent a novel species of the genus Methanocella, for which the name Methanocella arvoryzae sp. nov. is proposed. The type strain is MRE50(T) (=NBRC 105507(T) =DSM 22066(T)).


Baseline characteristics and changes of biomarkers in disease course predict prognosis of patients with COVID-19.
The outbreak of coronavirus disease (COVID-19) has brought great challenges to the world. The objectives of this study were to describe the baseline characteristics and changes of biomarkers of these COVID-19 patients and identify predictive value of the above markers for patient death. Using patient death as the observational endpoints, clinical data of inpatients in a special ward for COVID-19 in Wuhan, China were retrospectively collected. Univariate and multivariate Cox regression analyses were used to evaluate prognostic value of baseline characteristics and laboratory data changes. This study included clinical data of 75 patients. Age, c-reactive protein (CRP) and interleukin-6 levels were independent predictors of patient death. Survivors were characterized as having declining neutrophil counts, D-dimer, N-terminal pronatriuretic peptide, troponin I (TnI) and c-reactive protein levels, while counts of lymphocyte gradually came back. Non-survivors were characterized with increasing white blood cell counts (WBC) and neutrophil counts. Changes of WBC, TnI and interleukin-6 were also independently associated with patient death. Older age, baseline CRP and IL-6 levels may be used as meaningful predictors to identify patients with poor prognosis. Changes of biomarkers should be closely monitored in the management of patients with COVID-19, while constantly increasing levels of WBC, TnI and interleukin-6 in the disease course also predict patient death.


Microbial gene expression during hibernation in arctic ground squirrels: greater differences across gut sections than in response to pre-hibernation dietary protein content.
Obligate seasonal hibernators fast for 5-9 months depending on species yet resist muscle atrophy and emerge with little lean mass loss. The role of the gut microbiome in host nitrogen metabolism during hibernation is therefore of considerable interest, and recent studies support a role for urea nitrogen salvage (UNS) in host-protein conservation. We were interested in the effect of pre-hibernation diet on UNS and the microbial provision of essential amino acids (EAAs) during hibernation; therefore, we conducted a study whereby we fed arctic ground squirrels (Urocitellus parryii) pre-hibernation diets containing 9% vs. 18% protein and compared the expression of gut bacterial urease and amino acid (AA) metabolism genes in 4 gut sections (cecum mucosa, cecum lumen, small intestine [SI] mucosa, and SI lumen) during hibernation. We found that pre-hibernation dietary protein content did not affect expression of complete bacterial AA pathway genes during hibernation; however, several individual genes within EAA pathways were differentially expressed in squirrels fed 18% pre-hibernation dietary protein. Expression of genes associated with AA pathways was highest in the SI and lowest in the cecum mucosa. Additionally, the SI was the dominant expression site of AA and urease genes and was distinct from other sections in its overall microbial functional and taxonomic composition. Urease expression in the gut microbiome of hibernating squirrels significantly differed by gut section, but not by pre-hibernation dietary protein content. We identified two individual genes that are part of the urea cycle and involved in arginine biosynthesis, which were significantly more highly expressed in the cecum lumen and SI mucosa of squirrels fed a pre-hibernation diet containing 18% protein. Six bacterial genera were responsible for 99% of urease gene expression: Cupriavidus, Burkholderia, Laribacter, Bradhyrizobium, Helicobacter, and Yersinia. Although we did not find a strong effect of pre-hibernation dietary protein content on urease or AA metabolism gene expression during hibernation, our data do suggest the potential for pre-hibernation diet to modulate gut microbiota function during hibernation, and further investigations are warranted.


Selective autophagy maintains centrosome integrity and accurate mitosis by turnover of centriolar satellites.
The centrosome is the master orchestrator of mitotic spindle formation and chromosome segregation in animal cells. Centrosome abnormalities are frequently observed in cancer, but little is known of their origin and about pathways affecting centrosome homeostasis. Here we show that autophagy preserves centrosome organization and stability through selective turnover of centriolar satellite components, a process we termed doryphagy. Autophagy targets the satellite organizer PCM1 by interacting with GABARAPs via a C-terminal LIR motif. Accordingly, autophagy deficiency results in accumulation of large abnormal centriolar satellites and a resultant dysregulation of centrosome composition. These alterations have critical impact on centrosome stability and lead to mitotic centrosome fragmentation and unbalanced chromosome segregation. Our findings identify doryphagy as an important centrosome-regulating pathway and bring mechanistic insights to the link between autophagy dysfunction and chromosomal instability. In addition, we highlight the vital role of centriolar satellites in maintaining centrosome integrity.


Primary laryngeal lymphoma.
Primary laryngeal lymphoma is a very rare entity, with fewer than 50 cases reported in the English literature in the past 60 years. Close scrutiny of some of these case reports reveals that the larynx was not always the only site of involvement, thereby diminishing the total number of patients with primary laryngeal lymphoma to fewer than 35. The authors report a series of six patients, who were seen and evaluated at the Mayo Clinic between 1952 and 1995, with stage IAE non-Hodgkin's lymphoma of the larynx. Three patients had large-cell lymphomas according to the REAL (Revised European-American Lymphoid) classification. The other three had a small lymphocytic lymphoma, follicular small cleaved lymphoma, and follicular mixed lymphoma. All patients received radiation therapy alone as initial therapy for their disease and all patients had a complete remission to initial therapy. Four patients subsequently relapsed and the histology at relapse was the same as the initial histology in all four patients. Five patients have died, three of lymphoma, with a median survival of 67 months (range, 40 to 228 months). In view of the heterogeneity of histologies in this group of lymphomas, the variability in duration of response, and the significant number of patients who died of their disease, it is more likely that primary laryngeal lymphoma is an unusual presentation of non-Hodgkin's lymphoma than a separate disease entity. Despite the small number of patients in this study, the data would suggest that patients are best treated according to the histology of the lymphoma, rather than the limited stage and location of the disease.


Sex differences in response to ketamine as a rapidly acting intervention for treatment resistant depression.
While ketamine has been increasingly studied for treatment resistant depression (TRD), the impact of sex differences on treatment outcomes has not been well studied. The objective was to ascertain whether there were differences in response to a single administration of ketamine for TRD between men and women, and between pre- and post-menopausal women. A randomized, double-blind, placebo-controlled trial (N = 99; N = 50 male; N = 49 female) was conducted to investigate the efficacy of intravenous ketamine versus active placebo as augmentation of antidepressant therapy for TRD. Patients were assigned to one of five arms; one-time administration of ketamine of varying doses (i.e., 0.1, 0.2, 0.5, and 1.0 mg/kg), and one group receiving active placebo (intravenous midazolam). A priori-planned analyses were conducted to compare responses between women and men, as well pre-vs. postmenopausal women. Analyses demonstrated no significant differences between women and men in terms of treatment response (F(1,80) = 0.06, p = 0.80). There were no significant differences in the frequency of adverse effects (AEs) reported by those assigned to ketamine treatment groups (p > 0.21 for all AEs reported more than once), although women reported more headaches (12% vs. 6%, p = 0.30) and nausea (10% vs. 6%, p = 0.47). In comparing pre-vs. postmenopausal women, no differences in efficacy were observed (F(1,76) = 0.36, p = 0.55). Results do not support differential efficacy or tolerability of ketamine for the treatment of TRD between women and men, nor based on menopause status among women. However, larger trials with these a priori aims are needed to confirm these results.


Childhood abuse affects emotional closeness with family in mid- and later life.
Knowledge about the effects of early life adversity on kin relationships in later years is sparse. The purpose of this study was to examine if childhood abuse and adversity negatively influences emotional closeness with family in mid- and later life. A second goal was to determine the role of psychosocial resources and personality traits in buffering the effects of early adversities. Gender and cohort differences were explored to see if men were differentially affected than women and whether middle-aged adults (35-49 years old) were differentially affected than older adults (50-74 years old) by the effects of childhood abuse and adversity. Using retrospective accounts of early family abuse and adversities of 1,266 middle aged adults and 1,219 older adults from a large population-based survey, the National Survey of Midlife Development in United States (MIDUS), separate multiple regression analyses were conducted for the two cohorts to examine the effects of childhood emotional and physical abuse and family adversities on perceived emotional closeness with family. Interaction effects between childhood abuse and adversity (e.g., being expelled from school, death of sibling, parental divorce, losing a home to a natural disaster) with psychosocial resources (perceived control and self acceptance), personality characteristics (extraversion and neuroticism), and gender were examined. Results of OLS regressions suggest emotional and physical abuse predicted family closeness in middle-aged adults. Conversely, only emotional abuse predicted family closeness in older adults. Moderation models revealed that high levels of self acceptance were associated with better maintenance of emotional closeness among middle-aged adults who were emotionally and physically abused as children. Older adults with lower extraversion who experienced emotional abuse or reported greater number of adversities in childhood were found to be at higher risk for lower emotional closeness with family. Early life adversities were more detrimental for women. Findings suggest that the aftermath of childhood abuse does not dissipate with time, but continues to influence family relationships in mid- and later life. Identifying the links between childhood adversities and adult relationships can help identify strategic points for intervention to reduce the long-term effects of accumulated adverse experiences over the life course.


Correlates of time spent walking and cycling to and from work: baseline results from the commuting and health in Cambridge study.
Environmental perceptions and psychological measures appear to be associated with walking and cycling behaviour; however, their influence is still unclear. We assessed these associations using baseline data from a quasi-experimental cohort study of the effects of major transport infrastructural developments in Cambridge, UK. Postal surveys were sent to adults who travel to work in Cambridge (n = 1582). Questions asked about travel modes and time spent travelling to and from work in the last week, perceptions of the route, psychological measures regarding car use and socio-demographic characteristics. Participants were classified into one of two categories according to time spent walking for commuting ('no walking' or 'some walking') and one of three categories for cycling ('no cycling', '1-149 min/wk' and ' ≥ 150 min/wk'). Of the 1164 respondents (68% female, mean (SD) age: 42.3 (11.4) years) 30% reported any walking and 53% reported any cycling to or from work. In multiple regression models, short distance to work and not having access to a car showed strong positive associations with both walking and cycling. Furthermore, those who reported that it was pleasant to walk were more likely to walk to or from work (OR = 4.18, 95% CI 3.02 to 5.78) and those who reported that it was convenient to cycle on the route between home and work were more likely to do so (1-149 min/wk: OR = 4.60, 95% CI 2.88 to 7.34; ≥ 150 min/wk: OR = 3.14, 95% CI 2.11 to 4.66). Positive attitudes in favour of car use were positively associated with time spent walking to or from work but negatively associated with cycling to or from work. Strong perceived behavioural control for car use was negatively associated with walking. In this relatively affluent sample of commuters, a range of individual and household characteristics, perceptions of the route environment and psychological measures relating to car use were associated with walking or cycling to and from work. Taken together, these findings suggest that social and physical contexts of travel decision-making should be considered and that a range of influences may require to be addressed to bring about behaviour change.


Scanning electron microscopic observations of common carotid artery endothelium in the rat. II. Sutured arteries.
Scanning electron microscopic findings in 26 rats with sutured 1 to 1.2 mm segments of carotid artery indicate a difference in morphological changes beneath clamped areas and areas 2 to 3 mm adjacent to the suture line. Craters similar to those observed in control specimens were seen beneath clamped sites without significant platelet aggregation or platelet adherence to the endothelial wall. In contrast, areas adjacent to the suture line exhibited a pattern of fibrin and platelet adherence to the vessel endothelial wall. These changes appeared to be consistent after one to four hours of observation in most animals. Sutures, although attracting some platelets, did not exhibit platelet and fibrin adherence as marked as that of the adjacent endothelial wall.