The mechanism of FZXJJZ decoction suppresses colorectal liver metastasis via the VDR/TGF-β/Snail1 signaling pathways based on network pharmacology-TCGA data-transcriptomics analysis.
Fuzheng Xiaojijinzhan (FZXJJZF) decoction is an effective prescription for treating colorectal cancer liver metastasis (LMCRC). To elucidate the pharmacological mechanism of the FZXJJZF decoction therapy on LMCRC. Firstly, a network pharmacological approach was used to characterize the underlying targets of FZXJJZF on LMCRC. Secondly, LMCRC-related genes are obtained from the public database TCGA, and those genes are further screened and clustered through Mfuzz, an R package tool. Then, targets of FZXJJZF predicted by network pharmacology were overlapped with LMCRC related genes screened by Mfuzz. Meanwhile, FZJZXJF intervened in LMCRC model,epithelial-to-mesenchymal transition (EMT), and migration and invasion of HCT-116 cells. Thirdly, the transcriptomics data of FZJZXJF inhibited HCT-116 cells of EMT cells were overlapped with EMT database data to narrow the possible range of targets. Based on this, the potential targets and signal pathways of FZJZXJF were speculated by combining the transcriptomics data with the targets from network pharmacology-TCGA. Finally, the anti-cancer mechanism of FZXJJZF on LMCRC was verified in vitro by Real-Time PCR and Western Blot in vitro. By network pharmacological analysis, 282 ingredients and 429 potential targets of FZXJJZF were predicted. The 9268 LMCRC-related genes in the TCGA database were classified into 10 clusters by the Mfuzz. The two clustering genes with the most similar clustering trends were overlapped with 429 potential targets, and 32 genes were found, such as CD34, TRPV3, PGR, VDR, etc. In vivo experiments, FZJZXJF inhibited the tumor size in LMCRC models, and the EMT, migration, and invasion of HCT-116 also be inhibited. Intersecting transcriptomics dates with 32 target genes, it is speculated that the VDR-TGF-β signaling pathway may be an effective mechanism of FZXJJZF. Additionally, it is shown that FZXJJZF up-regulated the expression levels of VDR and E-cadherin and down-regulated the expression levels of TGF-β and Snail1 in vitro. These results confirmed that FZXJJZF plays an effective role in LMCRC mainly by inhibiting EMT phenotype via the VDR-TGF-β signaling pathway. Collectively, this study reveals the anti-LMCRC effect of FZXJJZF and its potential therapeutic mechanism from the perspective of potential targets and potential pathways.


Study of anorectal dynamics in patients undergoing laparoscopic ultra-low resection and transanal intersphincteric resection for rectal cancer.
Quite a few studies on anal functions after open total mesorectal excision combined with transanal intersphincteric resection (ISR) have been reported, but there is little literature on anal function after laparoscopic total mesorectal excision (LTME) combined with transanal ISR. The aim of this study was to explore the post-operative anorectal dynamic changes in ultra-low rectal cancer patients undergoing LTME combined with transanal ISR. The data of 26 ultra-low rectal cancer patients undergoing LTME + transanal ISR were analysed. A total of 30 patients undergoing laparoscopic low anterior resection by the same surgeons during the same period were randomly enrolled into the control group. There were no differences in the preoperative anorectal manometry data and Wexner anal function scores between the observation group and the control group (P > 0.05). There were no significant differences in the mean operation time, the mean amount of bleeding and the mean post-operative hospital stay between the two groups (P > 0.05). The mean follow-up time was 16 months. No recurrence and metastasis were found in all cases. At 3 and 6 months after the operation, there were significant differences in the anorectal manometry data and Wexner anal function scores between the two groups (P < 0.05). However, at 1 year after the operation, there were no significant differences in the anorectal manometry data and Wexner anal function scores between the two groups (P > 0.05). Laparoscopic ISR for ultra-low rectal cancer is technically feasible, but the surgical indications should be strictly defined.


Analyses on lipid-rich organelles of the rat liver after bile duct ligation.
A fraction of lipid-rich organelles of rat livers was analyzed to study the effect of bile duct ligation on lipid metabolism. The lipid fraction of the control rats consisted mainly of Golgi-derived dense bodies. Three days after the ligation, myelin-like figures and increased phospholipids and cholesterol were characteristic of the fraction. Two weeks ligation resulted in proliferation of lysosomal electron dense bodies. As their volume density expanded, phospholipids and cholesterol increased. Electrophoresis indicated a reduced rate of VLDL assembly after ligation. The Golgi-dominant phase is converted to the lysosomal phase in the heavy subfraction after the bile duct ligation. This subcellular change could be a consequence of secondary lipidosis: hepatocyte lysosomes have been exhausted in sorting accumulated lipids into secretory lipoproteins.


Plasmodium's sticky fingers.
The life cycle of the malaria parasite (Plasmodium) is remarkably complex. Malaria parasites must engage in highly specific and varied interactions with cell types of both the mammalian host and the mosquito vector. In this issue of Cell, report detailed molecular insights into an intimate interaction between a malaria parasite protein and its host cell receptor that enables the parasite to invade erythrocytes.


Fiber reinforced hydrated networks recapitulate the poroelastic mechanics of articular cartilage.
The role of poroelasticity on the functional performance of articular cartilage has been established in the scientific literature since the 1960s. Despite the extensive knowledge on this topic there remain few attempts to design for poroelasticity and to our knowledge no demonstration of an engineered poroelastic material that approaches the physiological performance. In this paper, we report on the development of an engineered material that begins to approach physiological poroelasticity. We quantify poroelasticity using the fluid load fraction, apply mixture theory to model the material system, and determine cytocompatibility using primary human mesenchymal stem cells. The design approach is based on a fiber reinforced hydrated network and uses routine fabrication methods (electrohydrodynamic deposition) and materials (poly[ɛ-caprolactone] and gelatin) to develop the engineered poroelastic material. This composite material achieved a mean peak fluid load fraction of 68%, displayed consistency with mixture theory, and demonstrated cytocompatibility. This work creates a foundation for designing poroelastic cartilage implants and developing scaffold systems to study chondrocyte mechanobiology and tissue engineering. STATEMENT OF SIGNIFICANCE: Poroelasticity drives the functional mechanics of articular cartilage (load bearing and lubrication). In this work we develop the design rationale and approach to produce a poroelastic material, known as a fiber reinforced hydrated network (FiHy™), that begins to approach the native performance of articular cartilage. This is the first engineered material system capable of exceeding isotropic linear poroelastic theory. The framework developed here enables fundamental studies of poroelasticity and the development of translational materials for cartilage repair.


Combination of KIR and HLA gene variants augments the risk of developing birdshot chorioretinopathy in HLA-A*29-positive individuals.
Birdshot chorioretinopathy (BCR), a chronic ocular inflammatory disease with characteristic choroidal lymphocytic infiltrates, has been strongly associated with human leukocyte antigen (HLA)-A29. Although HLA-A29 occurs frequently in all populations, BCR affects only a small percentage of HLA-A29-positive Caucasians, indicating additional susceptibility factors for BCR. Discovery of HLA class I-specific killer cell immunoglobulin-like receptors (KIR) led to a series of epidemiological studies implicating KIR-HLA gene combinations in disease. Here, we characterized KIR-HLA pairs in BCR patients and controls carrying HLA-A*29 as well as controls lacking HLA-A*29. KIR-HLA pairs implicated for weak inhibition (KIR2DL2/3+HLA-C1 and KIR3DL1+HLA-Bw4(T80)) in combination with activating KIR genes associated with autoimmunity (KIR2DS2, 2DS3 and 2DS4) augment the risk of developing BCR in HLA-A*29-positive individuals. The reciprocal association of strong inhibitory pairs (KIR3DL1+HLA-Bw4(I80) and KIR2DL1+HLA-C2) in combination with those implicated in protection from infection (KIR3DS1+HLA-Bw4(I80) and KIR2DS1+HLA-C2) was observed in HLA-A*29-negative controls. These results suggest that a profound effect of KIR2DS2/S3/S4 in the absence of strong inhibition may enhance the activation of natural killer cells and T-cell subsets against intraocular self-antigens, thereby contributing to pathogenesis of BCR.


Carotenoid composition and metabolism in green and blue-green algal lichens in the field.
The carotenoid composition of 33 species of green algal lichens and 5 species of blue-green algal lichens was examined and compared with that of the leaves of higher plants. As in higher plants, green algal lichen species which were found in both shade and full sunlight exhibited higher levels of the carotenoids involved in photoprotective thermal energy dissipation (zeaxanthin as well as the total xanthophyll cycle pool) in the sun than in the shade. This was particularly true when thalli were moist during exposure to high light, or presumably became desiccated in full sunlight. However, the reverse trend in the carotenoid composition of green algal lichens was also observed in those species which were found predominantly either in the shade or in full sunlight. In this case sun-exposed lichens often possessed lower levels of zeaxanthin and of the components of the xanthophyll cycle than lichens which were found in the shade. In contrast to higher plants, the lichens from all habitats exhibited a relatively high ratio of carotenoids to chlorophylls (more characteristic of sun leaves), very low levels of α-carotene (similar to that found in sun leaves), and a level of β-carotene similar to that found in shade leaves. Zeaxanthin, but not the expoxides of the xanthophyll cycle, was also frequently found in blue-green algal lichens. A trend for increasing levels of zeaxanthin with increasing growth light regime was observed inPeltigera rufescens, the species which was found to occur over the widest range of light environments. The level of zeaxanthin per chlorophylla in these blue-green algal lichens was in a range similar to that per chlorophylla+b in green algal lichens. However, zeaxanthin was also absent in one species,Collema cristatum, in full sunlight. Thus, the zeaxanthin content of the blue-green algal lichens can be similar to that of higher plants, or it can be rather dissimilar, as was also the case in the green algal lichen species. The presence of large amounts of ketocarotenoids in blue-green algal lichens is also noteworthy.


Comparative kinetics of organophosphates and oximes with erythrocyte, muscle and brain acetylcholinesterase.
There is an ongoing debate whether oximes can effectively counteract the effects of organophosphorus compounds (OP) on brain acetylcholinesterase (AChE) activity and whether there are differences in the kinetic properties of brain and erythrocyte AChE. In order to investigate the kinetics of AChE from different tissues and species the well established dynamically working in vitro model with real-time determination of membrane-bound AChE activity was adapted for use with brain AChE. The enzyme reactor, that was loaded with brain, erythrocyte or muscle AChE, was continuously perfused with substrate and chromogen while AChE activity was on-line analyzed in a flow-through detector. It was possible to determine the Michaelis-Menten constants of human erythrocyte, muscle and brain AChE which were almost identical. In addition, the inhibition kinetics of sarin and paraoxon as well as the reactivation kinetics of obidoxime and HI 6 were determined with human, swine and guinea pig brain and erythrocyte AChE. It was found that the inhibition and reactivation kinetics of brain and erythrocyte AChE were highly comparable in all tested species. These data support the view that AChE from different tissue has similar kinetic properties and that brain AChE is comparably susceptible toward reactivation by oximes.


Neuraminidase inhibitory polyketides from the marine-derived fungus Phoma herbarum.
Two new polyketides, arthropsadiol C (1) and massarilactone H (2), together with six known derivatives (3-8) were isolated from the culture broth of the marine-derived fungus Phoma herbarum. Their structures were elucidated on the basis of spectroscopic methods, including 2D NMR techniques. Compounds 2, 4, 5, and 8 showed moderate neuraminidase inhibitory activity with IC(50) values ranging from 4.15 to 9.16 µM.


Dataset for the thermodynamic description for the NaF-KF-RbF-ZnF2 system.
This data article includes the supplementary database material for the manuscript "Thermodynamic description for the NaF-KF-RbF-ZnF2 system", (Tang et al., 2018). We present all the thermodynamic parameters used and optimized model parameters of the thermodynamic database of the NaF-KF-RbF-ZnF2 quaternary system, by which researchers can obtain the calculated phase diagrams. What׳s more, the comparison of calculations with experimental lattice constants of all compounds and relative deviations of unit cell parameters between before and after structure optimization for the NaF-KF-RbF-ZnF2 system are demonstrated in this work. Meanwhile, we provide the calculated mixing enthalpy in the binary AF-ZnF2 (A=Na, K and Rb) and AF-RbF (A=Na and K) system, which are helpful for researchers to better understand the system. These supplementary databases are most useful in understanding the manuscript (Tang et al., 2018).


True genes for human U1 small nuclear RNA. Copy number, polymorphism, and methylation.
The nucleotide sequence of a human U1 small nuclear RNA gene (HU1-1) is presented, along with several hundred nucleotides of its 5' and 3' flanking regions. We demonstrate that the immediate 5' flanking region sequences are a highly specific hydridization probe capable of distinguishing U1 RNA true genes from pseudogenes. The copy number of U1 RNA genes was determined to be constant, about 30/haploid genome equivalent in six different individuals. Nevertheless, several restriction enzyme cleavage site polymorphisms exist in these genes. We also show that C-C-G-G sequences in the vicinity of U1 RNA true genes are not methylated, but they are methylated near pseudogenes. Genomic DNAs of mouse, frog, chicken, and fruit fly contain sequences homologous to the human U1 RNA gene coding region but not to the 5' flanking region. This suggests that sequences important for expression of U1 RNA genes constitute only a minor part of the 5' flanking regions. In the HU1-1 locus, sequences similar to the consensus T-A-T-A-A-A box are located about 45 and 210 base pairs upstream of the point corresponding to the 5' end of the mature RNA. Several additional direct repeats which may be regulatory elements are present in the upstream region. A few short inverted repeats which probably constitute a transcription termination signal are in the region corresponding to the 3' end of the U1 RNA.


Moisture-triggered fast crystallization enables efficient and stable perovskite solar cells.
Understanding the function of moisture on perovskite is challenging since the random environmental moisture strongly disturbs the perovskite structure. Here, we develop various N2-protected characterization techniques to comprehensively study the effect of moisture on the efficient cesium, methylammonium, and formamidinium triple-cation perovskite (Cs0.05FA0.75MA0.20)Pb(I0.96Br0.04)3. In contrast to the secondary measurements, the established air-exposure-free techniques allow us directly monitor the influence of moisture during perovskite crystallization. We find a controllable moisture treatment for the intermediate perovskite can promote the mass transportation of organic salts, and help them enter the buried bottom of the films. This process accelerates the quasi-solid-solid reaction between organic salts and PbI2, enables a spatially homogeneous intermediate phase, and translates to high-quality perovskites with much-suppressed defects. Consequently, we obtain a champion device efficiency of approaching 24% with negligible hysteresis. The devices exhibit an average T80-lifetime of 852 h (maximum 1210 h) working at the maximum power point.


Tier-1 assays for assessing the toxicity of insecticidal proteins produced by genetically engineered plants to non-target arthropods.
In assessing an insect-resistant genetically engineered (IRGE) crop before its commercialization, researchers normally use so-called "Tier-1 assays" as the initial step to determine the effects of the crop on non-target organisms. In these tests, the insecticidal proteins (IPs) produced by the IRGEs are added to the diets of test organisms in the laboratory. Test organisms in such assays can be directly exposed to much higher concentrations of the test IPs than they would encounter in the field. The results of Tier-1 assays are thus more conservative than those generated in studies in which the organisms are exposed to the IPs by feeding on IRGE plant tissue or in the case of predators or parasites, by feeding on invertebrate prey or hosts that have fed on IRGE plant tissue. In this report, we consider three important factors that must be considered in Tier-1 assays: (i) methods for delivery of the IP to the test organisms; (ii) the need for and selection of compounds used as positive controls; and (iii) methods for monitoring the concentration, stability and bioactivity of the IP during the assay. We also analyze the existing data from Tier-1 assays regarding the toxicity of Bt Cry proteins to non-target arthropod species. The data indicate that the widely used Bt proteins have no direct toxicity to non-target organisms.


Impact of Hemodynamic Instability and Organ Malperfusion in Elderly Surgical Patients Treated for Acute Type A Aortic Dissection.
Recent studies indicate acceptable survival rates in elderly patients treated surgically for acute type A aortic dissection (aTAAD). However, the impact of preoperative hemodynamic compromise or organ malperfusion on outcomes of such patients is still unclear. In a retrospective study of 341 patients, 101 qualified as elderly (≥70 years old). Subjects were further grouped by clinical presentation, using the Penn classification. Univariate and multivariable analyses were conducted to identify variables reflecting in-hospital and long-term mortality. Relative to younger subjects, elderly patients showed significantly higher rates of in-hospital mortality (24.8% vs. 14.6%, p = 0.025) and DeBakey type 2 dissections at presentation (40% vs. 18% p < 0.001), with significantly fewer presenting as Penn class Ab (p = 0.010). Penn class Ac was identified as an independent predictor of in-hospital mortality at all ages. Estimated long-term survival was poorer in the elderly (log rank p < 0.001); but in-hospital mortality, based on Penn classification, was similar for both age groups. Survival rates of Penn class Aa subjects at one, five, and 10 years were lower in elderly (vs. younger) patients (79 ± 5.6% vs. 90 ± 2.7%, 68 ± 6.7% vs. 80 ± 3.9%, and 39 ± 10.3% vs. 75 ± 4.6%, respectively; log rank p < 0.001). Overall in-hospital mortality is higher in elderly patients surgically treated for aTAAD. Malperfusion and/or hemodynamic instability at presentation confer a dismal prognosis, independent of patient age.


Molecular cloning and functional analysis of a novel 6&1-FEH from wheat (Triticum aestivum L.) preferentially degrading small graminans like bifurcose.
Like barley and other cereals, wheat (Triticum aestivum L.) accumulates branched graminan-type fructans containing both beta-(2,1) and beta-(2,6) fructosyl linkages, mainly with a quite low degree of polymerization (DP). 1&6-kestotetraose (bifurcose) is the major fructan oligosaccharide accumulating in crown tissues and leaves of cereals exposed to chilling. The fructan exohydrolase (FEH) cDNAs 1-FEH w1 and w2 were previously cloned from wheat crowns sampled in mid-November. Here, we report the cloning and functional analysis of another FEH cDNA from a mid-November wheat crown cDNA library. The cDNA encodes a long open reading frame (ORF) of 595 amino acids. Like other FEHs, it has a low iso-electric point (5.2) and it groups together with cell-wall type invertases and not with vacuolar invertases. The deduced amino acid sequence shows 67% identity to wheat 1-FEH w1 and w2. Functional characterization of the recombinant proteins in Pichia pastoris demonstrated that the recombinant enzyme had FEH activity towards the pure compounds 1-kestose, 6-kestose, 1,1-nystose and 1,1,1-kestopentaose. However, when incubated with its putative natural substrates (a mixture of low DP graminans from wheat crowns), it was shown that 1&6-kestotetraose (bifurcose) was preferentially removed from the graminan mixture. High DP wheat graminan and bacterial levan were only poor substrates. No hydrolase activities could be detected towards sucrose and high DP inulin, convincingly demonstrating that the enzyme is not a classic invertase or 1-FEH. The enzyme was termed 6&1-FEH w1. Northern blot analyses showed that 6&1-FEH w1 was expressed in crown tissue from autumn through winter under snow, while the expression levels in leaves were minimal or not detectable. The results strongly suggest that this unique FEH might play an important role in the degradation of branched, low DP wheat graminan (like bifurcose) in wheat crowns in the high fructan content season.


A putative role for colloidal silicates in primitive evolution deduced in part from their relevance to modern pathological afflictions.
Biogenic and bioactive silicas have structural subtleties that are only now beginning to be appreciated. Their ability to interact with particular biological systems is related to these structures. Some of these interactions result in pathological effects. It is suggested that such interactions mirror events occurring in a prebiotic environment.


Relationship of body water compartments to age, race, and fat-free mass.
Water compartments were studied in 72 black and 128 white women, aged 20 to 70 years. Total body water (TBW) was measured by tritiated water dilution, and extracellular water (ECW) was measured by using delayed gamma neutron activation for the determination of total body chloride. Intracellular water (ICW) was assessed as the difference between TBW and ECW. Fat-free mass (FFM) was estimated by the measurement of total body potassium (TBK) and total body fat (TBF) by measurement of total body carbon (TBC). Total body protein was calculated from total body nitrogen (TBN). TBW was found to decline with age and to also be significantly influenced by body size. The extracellular water space was 41.5% of TBW. The decline of TBW with age is due primarily to a decline in ICW. The hydration of the FFM increased with age for the overall population because of a decline in TBK and TBN and an increase in ECW. Body composition models that have assumed constancy of hydration should be adjusted not only for gender but for age and body size.


Phenotype Discordance between Primary Tumor and Metastasis Impacts Metastasis Site and Outcome: Results of WSG-DETECT-PriMet.
Tumor biological factors of breast cancer (BC) such as hormone receptor (HR) status, HER2 status, and grade can differ in the metastatic cascade from primary to lymph node (LN) metastasis and to distant metastatic tissue. Systematic data regarding therapeutic consequences are yet limited. We conducted a prospectively planned, retrospective cohort study comparing BC phenotype in tissue from primary tumors (PTs), locoregional LN metastases, and disease recurrence (DR). HR and HER2 as well as tumor grade in PTs and DR were obtained by a database search. No centralized biomarker testing was performed. The impact of changes in tumor biological factors on post-recurrence survival (PRS) and overall survival was analyzed. PriMet comprises 635 patients (LN tissue in 142 patients). Discrepancies for HR or HER2 status between PT and DR were observed in 18.7 and 21.6% of cases, respectively. For HR status, positivity of PT and negativity of DR was seen more often (13.2%) than vice versa (5.5%). For HER2 status, negativity of the primary and positivity of DR was seen more often (14.9%) than vice versa (6.7%). Discordance was more often observed between PT and LN metastasis compared to LN versus DR. However, numbers were small. Compared to concordant non-triple-negative (TN) disease, concordant TN disease showed significantly inferior PRS. We demonstrate receptor discordance to occur relatively frequently between PT, LN metastasis, and DR and to impact patient prognosis. However, clinical consequences of receptor discordance need to be drawn with caution considering clinical aspects as well as tumor biology.


Ventilation-perfusion lung imaging in nitrofurantoin-related pulmonary reaction.
A woman with a history of multiple drug allergies was admitted for cough and dyspnea. She had been taking nitrofurantoin for 10 days, was febrile, and had a rash on the trunk and extremities. A chest radiograph revealed right-sided pleural effusion and basal markings. Because of suspected nitrofurantoin-related pulmonary reaction, the medication was discontinued and the patient was started on an H2-blocker and intravenous corticosteroids. She improved clinically, and within 24 hours a radiograph revealed that her chest had largely cleared. Restricted distribution of radioxenon, seen initially on a ventilation lung scan, also reverted to near normal by the follow-up. The first perfusion lung scan showed multiple but nonspecific changes; this also came back to near normal. The nitrofurantoin-related pulmonary reaction should be considered in patients who present with pulmonary signs/symptoms while on the medication.


The pleiotypic response to amino acid deprivation is the result of interactions between components of the glycolysis and protein synthesis pathways.
Several diverse metabolic events become compromised when mammalian cells are made deficient in essential amino acids or when charging of their tRNA is blocked by amino acid analogs. This rapid general demise of cell function can be due to inhibition of phosphofructokinase (PFK) by uncharged tRNA. It has now been demonstrated that when tRNA is added to PFK in an assay dependent upon the reassociation of inactive, dissociated enzyme subunits, nanomolar concentrations cause complete inhibition. The model for control suggests that charged tRNA becomes associated with EF-1, which is specific for aminoacyl-tRNAs and is present in sufficiently high concentrations in cells to sequester that charged forms from an inhibitory role. Support for this model include: (1) the rapid onset of inhibition of glycolysis and glucose uptake upon amino acid deficiency; (2) the unique role of the product of PFK activity, fructose-1,6-diphosphate, in reactions of peptide chain initiation, particularly its role as a co-factor for purified eIF-2B, the GDP/GTP exchange factor; (3) the correlations of this interaction with the cellular and molecular lesions of insulin insufficiency; (4) the recognition that the anomalous role of high concentrations of cAMP as a stimulant of peptide chain initiation in energy depleted or gel-filtered cell lysates correlates with its stimulatory action on PFK as an analog for the positive effector, adenosine-5'-monophosphate; and (5) the role of fructose-1,6-diphosphate in the formation of glyceraldehyde-3-phosphate, a substrate for synthesis of ribose-5-phosphate via the non-oxidative portion of the pentose phosphate pathway, which, as a precursor of phosphoribosylpyrophosphate, is essential for nucleic acid synthesis.


Audit of nutritional advice given to bone marrow transplant patients in the United Kingdom and Eire.
To determine the dietary advice, catering practices, anthropometry and nutritional support given to bone marrow transplant (BMT) patients in the United Kingdom and Eire and to compare current advice with published literature recommendations. Questionnaire survey completed by consultants and dietitians. 43 hospitals undertaking autografts and/or allografts. Consultant Haematologists and Senior Dietitians. A 75% response rate was achieved. There is a wide variation in both dietary advice and nutritional support given to BMT patients. The wide variations in clinical practice reflect the lack of research in this area. The current problems of giving advice are discussed and some specific suggestions are made. Advice needs to be consistent and within the limitations of existing research.


Meal timing and subjective sleep disturbances in older men.
Older adults often complain about sleep disturbances, such as difficulty falling asleep and difficulty maintaining sleep in the early morning hours. Here, we investigated whether meal timing is associated with sleep problems in a cohort of older Swedish men (n = 998, mean age 71). Each participant filled out a seven-day food diary used to determine the daily eating time window, daily eating midpoint, and meal timing variability (i.e., the variance in daily eating midpoints over seven days). Questionnaires were used to assess difficulty initiating sleep and difficulty maintaining sleep. As indicated by logistic regression adjusted for potential confounders (e.g., BMI, diabetes status), no significant associations were found between the meal timing parameters and subjective sleep problems (P ≥ 0.37). Similar results were obtained when restricting the analysis to adequate reporters of daily energy intake. Therefore, our findings suggest that meal timing variations do not contribute to subjective sleep problems in older men. Our results must be replicated in cohorts that also include women and other measures of sleep.


[Inducement and proliferation of MSCs in vitro and preliminary study on interface between MSCs and metal materials].
The aims of the present study were to assess the characteristics of MSCs which were induced and proliferated in vitro, then to learn the interfaces between them and metal materials. Adult dogs were selected as experimental animals, the MSCs (marrow stem cells) were aspirated from the femur, which were induced to the phenotype of OB (osteoblast), then to proliferated in vitro with metal materials. Using biochemical tests, phase contrast microscopy, transmit electromicroscope, scanning electromicroscope to detect MSCs and the image of interface. MSCs had powerful osteogenic phenotype when they were induced in vitro, and they also attached to the metal materials. And there were more cells which attached on tensile stress area than other areas. The MSCs induced in vitro could be used as source of osteoblast. The affects of air--abrasion with alumina are not significant, but they have relation to the extent of strain and property of stress.


Deoxynivalenol and 3-acetyldeoxynivalenol - mycotoxins associated with wheat head fusariosis in Poland.
Samples of wheat naturally infected in the field byFusarium culmorum (W.G.Sm.) Sacc. andFusarium graminearum Schwabe were analyzed for deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone. No zearalenone was detected at levels higher than 0.5mg/kg. Deoxynivalenol was present in 100% and 3-acetyldeoxynivalenol in 80% of the examined samples at levels of 0.21 to 30.4 mg/kg and 0.54 to 29.54 mg/kg, respectively. The mycotoxin levels in the chaff were 5 to 50 times higher than in the kernels. This is the first report on natural occurrence of 3-acetyldeoxynivalenol in wheat. This toxin, in addition to deoxynivalenol, was highly correlated with wheat head fusariosis. These findings suggest that more attention should be given to the occurrence of 3-acetyldeoxynivalenol in cereal grains during the growth as well as during storage.


Improved renal clearance and tumor targeting of 99mTc-labeled anti-Tac monoclonal antibody Fab by chemical modifications.
This study was undertaken to improve the renal clearance and tumor targeting properties of 99mTc-labeled humanized anti-Tac (HuTac) monoclonal antibody Fab fragments using two chemical approaches: 1) labeling with a renal secretion agent 99mTc-mercaptoacetyltriglycine (MAG3) and 2) lowering its isoelectric point (pI) by acylation. HuTac Fab (3.3 mg/mL) was reacted with a trifluorophenyl ester (TFP) of 99mTc-MAG3 alone or was additionally reacted with TFP-glycolate to reduce the pI. In Balb/c mice, 99mTc-MAG3-Fab (pI > 9.3) rapidly accumulated in the kidneys (177% injected dose [ID]/g at 15 min) and then gradually cleared out of the kidneys. In contrast, the glycolation (pI 4.6 approximately 6.6) drastically reduced the renal uptake (31% ID/g) and also the whole-body retention (82% ID vs 101% for the nonglycolated) at 15 min, indicating that the glycolated 99mTc-MAG3-Fab (pI 4.6 approximately 6.6) was rapidly excreted. The glycolated remained in the blood longer than the nonglycolated (1.2% vs 0.3% ID/g at 360 min), but this effect was less drastic than the effect shown on the renal uptake. In nude mice bearing receptor-positive (ATAC4) tumors, the glycolated 99mTc-MAG3-Fab increased the peak tumor uptake to 14.8% ID/g from 8.3% ID/g for 99mTc-MAG3-Fab, whereas the glycolation resulted in a drastic reduction of the renal uptake at 15 min. We demonstrated that the renal clearance and the tumor targeting of Fab could be optimized by chemical modifications.


[Sorption of bacteria by blood cells as an indicator of the reaction of animals to the action of different doses of gamma rays].
In experiments with mice, guinea pigs, and dogs, a study was made of the sorptive capacity of peripheral blood cells after different gamma-radiation doses with reference to living Escherichia coli cells. The sorptive capacity of blood cells was inhibited in exposed animals the inhibition being maximum during the first 3 days following irradiation. Homologous immunoglobulin administered to mice 24 h before irradiation prevented the diminution of the sorptive capacity of cells and stimulated it during the first week following irradiation.


Systematic fractionation of oligosaccharides of human immunoglobulin G by serial affinity chromatography on immobilized lectin columns.
Human immunoglobulin G is known to contain 16 different biantennary complex-type asparagine-linked sugar chains, each of which occurs in a nonsialylated, monosialylated, or disialylated form. These oligosaccharides can be separated into 14 fractions by sequential affinity chromatography with Aleuria aurantia lectin (AAL)-Sepharose, RCA120-WG003, and E4-phytohemagglutinin-agarose columns. Twelve of them were found to contain a single oligosaccharide, while the fraction which passed through all three columns was shown to contain two oligosaccharides, GlcNAc beta 1----2Man alpha 1----6(+/- GlcNAc beta 1----4) (GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4GlcNAcOT. The fraction, which bound to the AAL-Sepharose column and passed through the remaining two lectin columns, also contained two oligosaccharides, GlcNAc beta 1----2Man alpha 1----6(+/- GlcNAc beta 1----4) (GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4 (Fuc alpha 1----6)GlcNAcOT. These results indicated that serial affinity chromatography with the three lectin columns can be used effectively to detect changes in the sugar chains of IgG resulting from diseases such as rheumatoid arthritis.


BacMam Platform for Vaccine Antigen Delivery.
Recombinant baculo viruses based on Autographa californica multiple nuclear polyhedrosis virus carrying vertebrate cell active expression cassettes, so-called BacMam viruses, are increasingly used as gene delivery vectors for vaccination of animals against pathogens. Different approaches for generation of BacMams exist and a variety of transfer vectors to improve target protein expression in vivo have been constructed. Here we describe a use of transfer vector which contains an insect cell-restricted expression cassette for the green fluorescent protein and thus enables easy monitoring of BacMam virus rescue, fast plaque purification of recombinants and their convenient titer determination and which has been proven to be efficacious for gene delivery in vaccination/challenge experiments.


Despite CD4 cell count rebound the higher initial costs of medical care for HIV-infected patients persist 5 years after presentation with CD4 cell counts less than 350 μl.
We determined that for HIV patients presenting with CD4 cell counts less than 350 μl the initial higher costs of care persisted over 5 years. Fifty-nine percent of new patients between 1 April 1998 and 1 April 2003 had CD4 cell counts less than 350 μl. Mean first year total costs ($19 917 $Cdn) were 2.5 times higher than for presentations with CD4 cell counts more than 350 μl ($7840). Total annual costs of care subsequently decreased to $15 663 by year 5, but still remained higher ($8883) than those with CD4 cell counts more than 350 μl despite a median CD4 cell count increase from 134 to 464 μl.


The comparison of Th1, Th2, Th9, Th17 and Th22 cytokine profiles in acute and chronic HIV-1 infection.
The aim of this study was to compare cytokine expression on both gene and protein levels in acute and chronic phase of HIV type 1 (HIV-1) infection. Thirty four patients were enrolled for cytokine expression analysis on protein level in acute and chronic stage of HIV-1 infection. Using PCR array technology, expression of 84 cytokine genes was measured in 3 patients in acute and 3 patients in chronic stage of HIV-1 infection. Bead-based cytometry was used to quantify levels of Th1/Th2/Th9/Th17/Th22 cytokines. The results showed statistically significant increase of 13 cytokine gene expression (cd40lg, csf2, ifna5, il12b, il1b, il20, lta, osm, spp1, tgfa, tnfsf 11, 14 and 8) and downregulation of the il12a expression in chronic HIV type 1 infection. Concentrations of IL-10, IL-4 and TNF-α were increased in the acute HIV type 1 infection when compared to control group. During chronic HIV type 1 infection there was an increase of IL-10, TNF-α, IL-2, IL-6, IL-13 and IL-22 levels when compared to control group. Comparison of cytokine expression between two stages of infection showed a significant decrease in IL-9 concentration. This study showed changes in cytokine profiles on both gene and protein levels in different stages of HIV-infection.


Dystonia in Parkinson's disease: clinical and pharmacological features.
We studied the features of dystonia in 9 patients with untreated idiopathic Parkinson's disease and in 56 patients on sustained treatment with L-dopa. Dystonia was seen as an initial symptom in patients with both early- and late-onset Parkinson's disease and included action dystonia of the limbs and cranial dystonia. Although the coexistence of parkinsonism and dystonia suggests a common pathophysiology, antiparkinsonian drugs did not consistently influence dystonic spasms. L-dopa-induced dystonia was seen as an off-period, biphasic, or peak-dose phenomenon. Each type showed a distinctive pattern of localization of dystonic spasms, possibly reflecting neurochemical aspects of basal ganglia somatotopy. Neuropharmacological studies performed in 12 patients suggest that off-period dystonia is genuinely induced by L-dopa and best relieved by antiparkinsonian agents.


[Morphological characteristics of osteopetrosis].
Osteopetrosis is a rare inherited bone disorder characterized by increased bone density owing to failure in bone resorption by the osteoclasts. The disease is genetically and histologically heterogeneous with a wide spectrum of microscopic findings. The histology varies from cases with a total absence of osteoclasts to bone biopsies characterized by high numbers of enlarged multinucleated osteoclasts on a background of sclerotic cancellous bone with or without additional defect of mineralization of the bone matrix. Here we present typical cases of human osteopetrosis on the basis of bone biopsies with four distinct genotypes (mutations of TNFRSF11A, TCIRG1, CNCL7, KINDLIN-3 genes) and discuss genotype-phenotype relationships. Analyzing human bone biopsies of rare skeletal disorders might improve our understanding of bone metabolism with possible implications for the clinical management of other bone diseases.


Family satisfaction with intensive care unit communication during the COVID-19 pandemic: a prospective multicentre Australian study Family Satisfaction - COVID ICU.
Virtual communication has become common practice during the coronavirus disease 2019 (COVID-19) pandemic because of visitation restrictions. The authors aimed to evaluate overall family satisfaction with the intensive care unit (FS-ICU) care involving virtual communication strategies during the COVID-19 pandemic period. In this prospective multicentre study involving three metropolitan hospitals in Melbourne, Australia, the next of kin (NOK) of all eligible ICU patients between 1 July 2020 and 31 October 2020 were requested to complete an adapted version of the FS-ICU 24-questionnaire. Group comparisons were analysed and calculated for family satisfaction scores: ICU/care (satisfaction with care), FS-ICU/dm (satisfaction with information/decision-making) and FS-ICU/total (overall satisfaction with the ICU). The essential predictors that influence family satisfaction were identified using quantitative and qualitative analyses. Seventy-three of the 227 patients' NOK who initially agreed completed the FS-ICU questionnaire (response rate 32.2%). The mean FS-ICU/total was 63.9 (standard deviation [SD], 30.8). The mean score for satisfaction with FS-ICU/dm was lower than the FS-ICU/care (62.1 [SD, 30.3) vs 65.4 (SD, 31.4); P < 0.001]. There was no difference in mean FS-ICU/total scores between survivors (n = 65; 89%) and non-survivors (n = 8, 11%). Higher patient Acute Physiology and Chronic Health Evaluation III score, female NOK and the patient dying in the ICU were independent predictors for FS-ICU/total score, while a telephone call at least once a day by an ICU doctor was related to family satisfaction for FS-ICU/dm. There was low overall family satisfaction with ICU care and virtual communication strategies adopted during the COVID-19 pandemic. Efforts should be targeted for improving factors with virtual communication that cause low family satisfaction during the COVID-19 pandemic.


In Vitro Antimicrobial Long-Term Evaluation of Corneal Preservation Media Against Microorganisms Using A Closed-Chamber Study Model.
To evaluate the existence of in vitro long-term antimicrobial activity of Optisol-GS against microorganisms related to corneal infection using a closed-chamber study model. Optisol-GS was contaminated with microorganisms related to corneal infections, and different times after contamination was analyzed using a closed-chamber study model. Microbial growths were analyzed by macroscopic observation. For Staphylococcus aureus and Pseudomonas aeruginosa, bacterial growth was observed in samples taken 1 hour through 7 days and 14 days after contamination occurred. For Staphylococcus epidermidis, Streptococcus agalactiae, and Candida albicans, microbial growth was observed in all samples studied. For Streptococcus pneumoniae, bacterial growth was observed in samples taken 1 hour through 72 hours after contamination. For Streptococcus pyogenes, bacterial growth was observed in samples taken 1 hour through 7 days after contamination. For Escherichia coli, bacterial growth was observed in samples taken 1 hour through 48 hours after contamination occurred. We conclude that no in vitro antimicrobial effect for any microorganism analyzed was observed in contaminated Optisol-GS after 72 hours; however, effective antimicrobial activity was observed for S. aureus, Str. pneumoniae, Str. pyogenes, P. aeruginosa, and E. coli after 7 to 10 days.


[Examining hypertension in children and adolescents: Clinical implications of the differences between the European and American Guidelines].
Over the past 2 years, new guidelines for hypertension in children and adults have been published. The first, developed by the European Hypertension Society Working Group on Hypertension in Children and Adolescents, recognised the need for an update of the of the 2009 European Guidelines of in order to incorporate the new information acquired over the last few years. Subsequently, an update of the American guidelines (the most recent of which was in 2004), has been published. In both guidelines there are aspects which are agreed on, and in others, there are marked differences in the basic elements that have an influence on daily clinical practice. The main differences are centred on the criteria for the diagnosis and classification of hypertension, with the subsequent impact on its prevalence. Future studies should offer responses to all the questions that still remain unresolved.


A female patient with adolescent-onset progressive myoclonus epilepsy carrying a truncating MECP2 mutation.
MECP2 is a well-known causative gene for Rett syndrome but other phenotypes have also been reported. Here, we report a case of a female patient with adolescent-onset progressive myoclonus epilepsy (PME) carrying a novel truncating mutation in the MECP2 gene. The patient was a 29-year-old woman with infantile-onset intellectual disability of unspecified cause. She had demonstrated slow but steady development with moderate intellectual disability until the age of 16, when she started having epileptic seizures. Her epilepsy progressed intractably with multiple seizure types accompanied by myoclonus, tremor, and gradual regression. She is currently apathetic and requires extensive assistance in all aspects of life. After an extensive work-up for underlying diseases for PME turned out negative, whole-exome sequencing revealed a de novo 113-bp deletion and 3-bp insertion in MECP2, a variant of NM_004992.4:c.1099_1211delinsGGG, p.(His367Glyfs*32). The clinical presentation of this case was inconsistent with Rett syndrome, and the rapid regression in the patient's twenties was considered characteristic. Mutations of MECP2 may result in variable neurodevelopmental phenotypes and may also be considered a causative gene for adolescent-onset PME.


Frailty affects prognosis in patients with colorectal cancer: A systematic review and meta-analysis.
The prevalence of colorectal cancer has remained high. Most patients have already developed into the middle and advanced stage when they are diagnosed with colorectal cancer, and a small number of them are accompanied by metastasis. In recent years, frailty has been recognized as an important factor affecting the prognosis of colorectal cancer. The aim of this study was to assess the value of frailty on prognosis in patients with colorectal cancer after treatment. We systematically searched PubMed, Embase, Web Of Science databases up until March2022. A total of 18 studies were retrieved that met the inclusion criteria, including 9 prospective studies and 9 retrospective studies. Frailty screening tools, proportion of frail patients, and outcomes of colorectal cancer patients after treatment were recorded. 18 studies were included with a total of 352,535 participants. Regardless of differences in frailty screening and treatment approaches, outcomes for frailty patients were less favorable in all studies. Compared with the non-frail group, the frail group had higher mortality, more serious complications, more postoperative blood transfusions and delirium, and more support outside the home. Although there is no uniform standard for frailty screening, assessing the frailty of colorectal cancer patients is of great significance for predicting prognosis of patients after treatment.


Comparative evaluation of the cerebral state index and the bispectral index during target-controlled infusion of propofol.
Cerebral state index (CSI) has recently been introduced as an intra-operative monitor of anaesthetic depth. We compared the performance of the CSI to the bispectral index (BIS) in measuring depth of anaesthesia during target-controlled infusion (TCI) of propofol. Twenty Chinese patients undergoing general anaesthesia were recruited. CSI and BIS, and predicted effect-site concentration of propofol were recorded. The level of sedation was tested by Modified Observer's Assessment of Alertness/Sedation Scale (MOAAS) every 20 s during stepwise increase (TCI, 0.5 microg ml(-1)) of propofol. The loss of verbal contact (LVC) and loss of response (LOR) were defined by MOAAS values of 2-3 and less than 2, respectively. Baseline variability and the prediction probability (P(K)) were calculated for the BIS and CSI. The values of BIS(05) and CSI(05), BIS(50) and CSI(50), BIS(95) and CSI(95) were calculated at each end-point (LVC and LOR). Baseline variability of CSI was more than that of BIS. Both CSI and BIS showed a high prediction probability for the steps awake vs LVC, awake vs LOR, and LVC vs LOR, and good correlations with MOAAS values. Despite larger baseline variation, CSI performed as well as BIS in terms of P(K) values and correlations with step changes in sedation.


Behavioral and electroencephalographic correlates of 40-Hz EEG biofeedback training in humans.
Two groups of eight adults successfully trained with biofeedback for increases in 40-Hz EEG responses in left or right hemispheres also demonstrated significant 40-Hz EEG increases during baseline periods, and increases in the contralateral hemisphere during training periods. No changes in heart rate, 40-Hz EMG, or 21- or 31-Hz beta, alpha, or theta EEG occurred over training days. Three subjects returning for additional training demonstrated suppression of 40-Hz EEG. A group of four subjects experiencing daily bidirectional training produced substantial within-session control of 40-Hz EEG but no changes over days. Data from posttraining tests without feedback for successful subjects in both groups indicated significant control of 40-Hz EEG responses in the initial parts of these sessions, and some correlated changes in other EEG responses. Measures of successful subjects' experiences during training and control tests indicated awareness of changes in subjective concomitants of EEG responses. This study suggests further strategies for research on behavioral correlates of EEG activity.


Physiological responses of Holstein calves to hot weather conditions.
The objectives of the study described were to (1) compare environmental temperature-humidity index (THI) with the THI measured within two different calf housing systems and (2) determine how THI affects Holstein heifer calf body temperatures, serum cortisol concentrations, and serum thyroxine concentrations. At 24 to 48 h of age, calves were assigned to one of two individual housing treatments: (1) stalls in a three-sided barn (n = 8) or 2) hutches placed outside (n = 8). Calves were observed until 42 days of age during the summer months. Ambient temperature and relative humidity within housing systems were recorded hourly. Calf body temperatures were measured hourly using devices attached to the underside of each calf's tail. Blood samples were collected at 7, 21, and 42 days of age via jugular venipuncture. Blood serum was analyzed for cortisol and thyroxine concentrations using enzyme-linked immunosorbent assays (ELISAs). Overall average THI was higher in hutches than in stalls (P < 0.0001). Calves housed in hutches had higher body temperatures (39 ± 0.18 °C vs 38 ± 0.15 °C, P = 0.005). No differences occurred in serum cortisol concentrations (P = 0.89) but at 21 days of age, calves housed in hutches had higher serum thyroxine concentrations (16.55 ± 0.51 μg/dl vs 13.91 ± 0.54 μg/dl, P = 0.0006) than calves housed in stalls. Although both housing systems in this study were located on the same dairy, calves housed in hutches were exposed to higher THI values, had higher body temperatures, and at 21 days of age had higher serum thyroxine concentrations than calves housed in stalls inside a three-sided barn.


Neuroblastoma: oncogenic mechanisms and therapeutic exploitation of necroptosis.
Neuroblastoma (NB) is the most common extracranial childhood tumor classified in five stages (1, 2, 3, 4 and 4S), two of which (3 and 4) identify chemotherapy-resistant, highly aggressive disease. High-risk NB frequently displays MYCN amplification, mutations in ALK and ATRX, and genomic rearrangements in TERT genes. These NB subtypes are also characterized by reduced susceptibility to programmed cell death induced by chemotherapeutic drugs. The latter feature is a major cause of failure in the treatment of advanced NB patients. Thus, proper reactivation of apoptosis or of other types of programmed cell death pathways in response to treatment is relevant for the clinical management of aggressive forms of NB. In this short review, we will discuss the most relevant genomic rearrangements that define high-risk NB and the role that destabilization of p53 and p73 can have in NB aggressiveness. In addition, we will propose a strategy to stabilize p53 and p73 by using specific inhibitors of their ubiquitin-dependent degradation. Finally, we will introduce necroptosis as an alternative strategy to kill NB cells and increase tumor immunogenicity.


Ultrastructure of astroglial contacts in rat cerebellar cortex.
Interfaces of the astroglial Bergmann fibres in the rat cerebellar cortex were investigated under the electron microscope. The straight interfaces revealed numerous contacts of a special junctional type having a mean length of 0.9 micro m. A substantial area of the Bergmann fiber surfaces was found to be connected with these junctions. Electron-dense material forming knoblike particles were observed attached with a certain regularity to the inner surfaces of both junctional membranes.


Selenium as an emerging versatile player in heterocycles and natural products modification.
The diverse pharmacological activities of organoselenium compounds are closely correlated to their ability to scavenge and induce reactive oxygen species (ROS), their intrinsic oxidative properties, and their Se(0) release property. The incorporation of selenium into small molecules, and particularly into heterocycles and natural products, has shown great potential in altering the potency and selectivity of these molecules. Therefore, selenium will play an important role in drug discovery in the near future. We summarize how different organoselenium species affect cellular oxidative stress levels, and try to correlate the structural properties of selenium-containing heterocycles and natural product derivatives to their biological activities and therapeutic applications. We also provide some information to guide the rational design of selenium-containing drugs.


Demystifying the Role of Natural Language Processing (NLP) in Smart City Applications: Background, Motivation, Recent Advances, and Future Research Directions.
Smart cities provide an efficient infrastructure for the enhancement of the quality of life of the people by aiding in fast urbanization and resource management through sustainable and scalable innovative solutions. The penetration of Information and Communication Technology (ICT) in smart cities has been a major contributor to keeping up with the agility and pace of their development. In this paper, we have explored Natural Language Processing (NLP) which is one such technical discipline that has great potential in optimizing ICT processes and has so far been kept away from the limelight. Through this study, we have established the various roles that NLP plays in building smart cities after thoroughly analyzing its architecture, background, and scope. Subsequently, we present a detailed description of NLP's recent applications in the domain of smart healthcare, smart business, and industry, smart community, smart media, smart research, and development as well as smart education accompanied by NLP's open challenges at the very end. This work aims to throw light on the potential of NLP as one of the pillars in assisting the technical advancement and realization of smart cities.


Can evaluation studies benefit from triangulation? A case study.
Information and communication technologies (ICTs) are increasingly being used in health care. Rigorous evaluations of ICT applications during both introduction and routine use are of great importance for decision makers and users. Within evaluation research, two main (and often rather distinct) traditions can be found: the objectivistic and the subjectivistic tradition. The theory of triangulation deals with the integration of methods and approaches as to conduct better evaluation studies. In evaluation research, triangulation in general means the multiple employment of various sources of data, observers, methods, and/or theories in investigations of the same phenomenon. We applied triangulation aspects in the analysis of the effects of a computer-based nursing documentation system. We discuss, based on this case study, what benefits can be obtained from applying triangulation in an evaluation study. We show how both the validation of results and the completeness of results can be supported by triangulation. The decision whether triangulation may be useful for a given research question, and how it may be correctly applied, requires-like other evaluation methods-intensive training and methodological experience. Medical informatics evaluation research may profit from this well-established theory.


Surgical techniques and results in pancreatic transplantation.
A major stumbling block preventing success with pancreatic transplantation has been the handling of the exocrine secretion from the pancreatic graft. As a consequence, numerous surgical techniques have been evaluated such as duct ligation, free intraperitoneal drainage, ductal filling with polymers, exocrine diversion to the urinary tract, to the bowel and to the stomach. Also, because of problems encountered with the duodenum, segmental body-and-tail pancreatic grafts have been used for many years. However, it seems today that mainly two different techniques for pancreatic transplantation remain. In both whole pancreaticoduodenal grafts are used with a vascular supply from the coeliac axis and the portal vein. The exocrine drainage is either to the urinary tract by means of a side-to-side anastomosis between donor duodenum and recipient urinary bladder, or to the bowel by a side-to-side anastomosis between donor duodenum and recipient proximal jejunum. With both these techniques the technical complications have been much reduced and as a consequence the graft survival rates are now approaching those obtained with the transplantation of the liver, heart and kidney.


Flow cytometry: A powerful analytical technique for characterizing the biological function of biotherapeutics and biosimilars.
Biotherapeutics are complex molecules with therapeutic activity produced through biotechnology and/or genetic engineering. These medicines have clinical applications in diagnostic procedures and therapies for many disorders, including cancer, autoimmunity, and chronic degenerative diseases. Most biotherapeutics are expensive and sometimes unaffordable for low-income patients suffering from cancer or chronic illness. Biosimilars emerged in the 2000 s after patents of many innovative biotherapeutic products expired. The Biosimilar market is growing fast and demands reliable technologies for analyzing the physicochemical properties and bioactivity of products. A big challenge for biosimilar development is to prove comparable bioactivity, safety, efficacy, and toxicity profile as the innovator product. Bioactivity assessment can utilize different analytical techniques such as ELISA, flow cytometry, and surface plasmon resonance. Flow cytometry is a versatile analytical tool that can be used for the development of quantitative, reproducible, and accurate protocols suitable for routine evaluation of bioactivity in-vitro. Nevertheless, flow cytometry has been very scarcely used in comparability evaluation between biosimilar versus an originator product. Here, we review potential applications of flow cytometry to carry out functional bioassays of biotherapeutics or biosimilars.


Co-digestion of grass silage and cow manure in a CSTR by re-circulation of alkali treated solids of the digestate.
Three laboratory, continuously stirred tank reactors (CSTRs) co-digesting grass silage and cow manure (forming 30% and 70% of substrate volatile solids (VS), respectively) were operated to evaluate the effects of re-circulating an alkali-treated and untreated solid fraction of the digestate back to the reactors. The CSTRs were operated at an organic loading rate (OLR) of 2 kg VS m(-3) day(-1) and hydraulic retention time (HRT) of 20 days with a semi-continuous mode of feeding. The feasibility of co-digestion with substrate VS containing 30% VS of crop was reinforced, resulting in average specific methane yield of about 180-185 1 CH4 kg(-1) VS. Re-circulation of the solid fraction of digestate back to the reactors in both alkali-treated and untreated forms decreased the methane yield by 11% and 21%, respectively, and resulted in operational problems such as scum formation and accumulation of the reactor materials. Batch studies were conducted to evaluate (i) the methane potentials of the solid fraction of digestate, and whole digestate with alkali treatments ranging from 20-60 g NaOH kg(-1) VS of substrate, and (ii) methane potentials of the accumulated reactor materials as top, middle and bottom layers. The solid fraction of digestate treated with 20 g NaOH kg(-1) VS showed higher specific methane yield (340 l CH4 kg(-1) VS) than the higher range of alkali treatments. The bottom layers of the control reactor and the reactor fed with alkali-treated solids gave a higher specific methane yield (93 and 85 l CH4 kg(-1) VS, respectively), and all three layers of untreated solids gave similar methane potentials.


Physiological relevance of the endogenous mono(ADP-ribosyl)ation of cellular proteins.
The mono(ADP-ribosyl)ation reaction is a post-translational modification that is catalysed by both bacterial toxins and eukaryotic enzymes, and that results in the transfer of ADP-ribose from betaNAD+ to various acceptor proteins. In mammals, both intracellular and extracellular reactions have been described; the latter are due to glycosylphosphatidylinositol-anchored or secreted enzymes that are able to modify their targets, which include the purinergic receptor P2X7, the defensins and the integrins. Intracellular mono(ADP-ribosyl)ation modifies proteins that have roles in cell signalling and metabolism, such as the chaperone GRP78/BiP, the beta-subunit of heterotrimeric G-proteins and glutamate dehydrogenase. The molecular identification of the intracellular enzymes, however, is still missing. A better molecular understanding of this reaction will help in the full definition of its role in cell physiology and pathology.


Characterization of covalent inhibitors that disrupt the interaction between the tandem SH2 domains of SYK and FCER1G phospho-ITAM.
RNA sequencing and genetic data support spleen tyrosine kinase (SYK) and high affinity immunoglobulin epsilon receptor subunit gamma (FCER1G) as putative targets to be modulated for Alzheimer's disease (AD) therapy. FCER1G is a component of Fc receptor complexes that contain an immunoreceptor tyrosine-based activation motif (ITAM). SYK interacts with the Fc receptor by binding to doubly phosphorylated ITAM (p-ITAM) via its two tandem SH2 domains (SYK-tSH2). Interaction of the FCER1G p-ITAM with SYK-tSH2 enables SYK activation via phosphorylation. Since SYK activation is reported to exacerbate AD pathology, we hypothesized that disruption of this interaction would be beneficial for AD patients. Herein, we developed biochemical and biophysical assays to enable the discovery of small molecules that perturb the interaction between the FCER1G p-ITAM and SYK-tSH2. We identified two distinct chemotypes using a high-throughput screen (HTS) and orthogonally assessed their binding. Both chemotypes covalently modify SYK-tSH2 and inhibit its interaction with FCER1G p-ITAM.


A contralateral eye study comparing characteristics of corneal endothelial cells in bilateral keratoconus patients with unilateral corneal Vogt's striae.
The aim of this study was to analyze and compare corneal endothelial cell morphology and characteristics in bilateral keratoconus (KCN) patients with unilateral Vogt's striae. Fifty patients aged 20-38 years were recruited in this cross-sectional contralateral eye study. In this study, corneal endothelial cell parameters were evaluated in patients with bilateral KCN and unilateral Vogt's striae using the Topcon SP2000P specular microscope (Topcon, Tokyo, Japan). In the current study, there were no significant differences in corneal endothelial cell parameters including endothelial cell density (ECD), hexagonal cell ratio (HEX), and coefficient of variance of cell size (CV) between the KCN groups with and without Vogt's striae, [(2968.34 ± 276.65 vs. 2980.05 ± 253.30, P = 0.618), (51.88 ± 13.57 vs. 53.24 ± 9.31, P = 0.658), and (32.50 ± 5.40 vs. 32.97 ± 4.07, P = 0.467), respectively]. Also, among study groups with and without Vogt's striae, ECD did not correlate with anterior chamber depth (ACD) [(P = 0.564, r = 0.09), (P = 0.219, r = -0.18), respectively], maximum keratometry (Kmax) [(P = 0.215, r = 0.18), (P = 0.898, r = 0.02), respectively], and central corneal thickness (CCT) [(P = 0.989, r = -0.02), (P = 0.643, r = -0.07), respectively].Our results showed significant differences in corrected and uncorrected distance visual acuity (UDVA), cycloplegic refractive error components (calculated by vectorial analysis), CCT, and Kmax between two study groups (all P < 0.05) except for J45 (Jackson cross cylinder, axes at 45 and 135°) (P = 0.131). We were not able to find the statistically significant differences in ECD, HEX, and CV between KCN eyes with and without Vogt's striae. Despite clinical and tomographic results, it seems that Vogt's striae cannot cause deterioration in the corneal endothelial morphology.


Reconditioning of heroin place preference requires the basolateral amygdala.
To investigate the role of the basolateral amygdala (BLA) in the reacquisition of heroin seeking, we studied the effect of BLA inactivation after heroin re-exposure in the presence of drug-conditioned cues. We employed a heroin conditioned place preference task [Leri F, Rizos Z, 2005. Reconditioning of drug-related cues: a potential contributor to relapse after drug re-exposure. Pharmacol Biochem Behav 2005;80:621-30.], where after initial conditioning and subsequent extinction, rats received a single reconditioning session (explicit compartment-heroin re-pairing), followed by a test of heroin seeking 24 h later. Rats were infused with GABA(A)/GABA(B) agonists (muscimol and baclofen, 0.03 and 0.3 nmol, respectively/0.3 microl) or vehicle, either 15 min or 6 h following the heroin reconditioning session. Animals that received vehicle infusions, whether they were given 15 min or 6 h following reconditioning, showed a significant preference for the heroin-paired compartment 24 h later. However, inactivation of the BLA 15 min post-reconditioning, but not 6 h following reconditioning, completely blocked the reacquisition of heroin seeking. These results suggest that the BLA plays an important role in a putative learning process initiated by drug re-exposure which may underlie the process of relapse.


Electrolyte economy and its hormonal regulation in congenital chloride diarrhea.
The economy of Cl-, K+, and Mg++, extracellular volume (ECV) and plasma volume, and the role of hyperreninemia and hyperaldosteronism were explored in 22 patients with congenital chloride diarrhea. Stool volume was in significant correlation with its Cl-, Na+ and K+ content, the correlation being significantly better with Cl- content than with the Na+ content. Low fecal Cl- concentrations were seen in chronic hypochloremic contraction, but acute episodes did not cause reduction of fecal Cl- concentration from the basal level of 140--150 mmol/liter. The adequate condition (defined as normal serum electrolyte concentrations and bl;od pH, and presence of Cl- in urine) was associated with high total exchangeable Cl- and ECV. This excess Cl- and ECV roughly equalled the high daily fecal amount of Cl- and volume. Reduced ECV was accompanied by high renin activities and hyperaldosteronism, but in the adequate condition these were normal. Hyperaldosteronism caused a decrease in urinary Na+-K+ ratio and, after the age of 2--6 months, in the fecal Na+-K+ ratio. Total exchangeable K+ was normal in the adequate condition. No Mg++ depletion was present, although the patients lack Mg++ substitution. The adequate condition could be maintained with an oral supplement of NaCl, KCl and water.


Loss of Borealin/DasraB leads to defective cell proliferation, p53 accumulation and early embryonic lethality.
Borealin/DasraB is a member of the chromosomal passenger protein complex (CPC) required for proper segregation of chromosomes during mitosis. In Drosophila melanogaster, inactivation of Borealin/DasraB results in polyploidy, delayed mitosis and abnormal tissue development, indicating its critical role for cell proliferation. However, the in vivo role of mammalian Borealin/DasraB remains unclear. Here, we analyzed the expression of Borealin/DasraB and found that borealin is widely expressed in embryonic tissues and later restricted to adult tissues which relies on rapid cell proliferation. To determine the role of borealin during mouse development, we generated borealin-null mice through targeted disruption. While heterozygous mice developed normally, disruption of both borealin alleles resulted in early embryonic lethality by 5.5 dpc (days postcoitus) due to mitotic defects and apoptosis in blastocyst cells that showed microtubule disorganization and no CPC enrichment. At 5.5 dpc, borealin-null embryos exhibited excessive apoptosis and elevated expression of p53. However, loss of p53 did not abrogate or delay embryonic lethality, revealing that Borealin/DasraB inactivation triggered impaired mitosis and apoptosis though p53-independent mechanisms. Our data show that Borealin/DasraB is essential for cell proliferation during early embryonic development, and its early embryonic lethality cannot be rescued by the loss of p53.


Risk factors and outcome of intra-abdominal hypertension in patients with severe acute pancreatitis.
Intra-abdominal hypertension (IAH) is common in patients with severe acute pancreatitis (SAP). The aim of the present study was to investigate the risk factors of IAH in SAP patients and assess the prognosis of SAP combined with IAH. To analyze the data from patients with SAP, both univariate and multivariate logistic regression analyses were applied, using 16 indices, including age, gender, Acute Physiology and Chronic Health Evaluation II scores (APACHE II), 24 h fluid balance, hematocrit, serum calcium level, and so on. Clinical prognosis such as mortality, hospital duration, of SAP patients with or without IAH was also compared. First 24 h fluid balance (Odds Ratio [OR], 1.003; 95% Confidence Interval [CI], 1.001-1.006), number of fluid collections (OR, 1.652; 95% CI, 1.023-2.956), and serum calcium level (OR, 0.132; 95% CI, 0.012-0.775) were found to be independent risk factors for IAH in patients with SAP. Moreover, patients with SAP and IAH had significantly longer average length of stay, both in the hospital and in the intensive care unit, higher rates of systemic and local complications, and more invasive treatments. The significant risk factors for IAH in patients with SAP include 24 h fluid balance (first day), number of fluid collections, and serum calcium level. Additionally, IAH is associated with extremely poor prognosis, evidenced by high rates of mortality, morbidity, and the need for invasive interventions.


New Zealand hospice nurses' self-rated comfort in conducting spiritual assessment.
To measure how comfortable hospice nurses in New Zealand are in conducting spiritual assessment and explore potentially associated factors. Cross-sectional, mixed-methods, descriptive study. Sixty hospice nurses from three New Zealand hospices. An investigator-designed questionnaire measured respondents' comfort with asking patients spiritual assessment questions proposed for use in the health-care literature. Other items assessed personal and work-related factors. Open questions generated data about how these nurses assess spirituality.The data was analysed using measures of central tendency, binomial non-parametric tests, and content analysis. These nurses were generally comfortable with proposed spiritual assessment questions and perceived spiritual assessment to be important. Personal factors including age, years in nursing, religiosity, and spirituality were not related to this comfort; however, training was associated with comfort. These findings underscore the importance of training and show how nurses can be comfortable with and capable of assessing patient spirituality.


Quality changes of Larimichthys crocea pretreated by ultrasonic-assisted immersion under cold storage at 4 °C.
The effects of several ultrasonic treatments during cold storage at 4 °C on the quality of large yellow croaker (Pseudosciaena crocea) were suggested. Large yellow croaker fillets were divided into six different groups for treatment. (a) CK (without any treatment), (b) A (single frequency 20 kHz), (c) B (single frequency 40 kHz), (d) C (left and right dual frequency 20 + 40 kHz), (e) D (orthogonal dual frequency: left and right 40 kHz, upper and lower 20 kHz), and (f) E (orthogonal dual frequency: left and right 20 kHz, upper and lower 20 kHz). The samples were divided into six groups, placed in sterile PE bags, and chilled to 4 °C. In order to determine the impact of ultrasonic treatment on the quality of large yellow croaker during cold storage, microbial indicators and physical and chemical indicators were measured every 3 days. The total number of colonies, the percentage of psychrophilic bacteria, the sample's pH, and its TVB-N value were all shown to grow at a much slower pace following ultrasonic treatment. In addition, the antibacterial effect of dual frequency ultrasound was gradually better than that of single frequency ultrasound. In conclusion, Group D has a pretty excellent impact on preserving overall sample quality.


Mercury-selenium interaction: distribution and excretion of 203Hg2+ in rats after simultaneous administration of selenite or selenate.
In female rats intravenously injected with 203HgCl2 (0.6 mg Hg2+ per kg body wt.) the effect of intraperitoneal administration of selenite or selenate (0.525 mg Se per kg body wt.) on distribution and excretion of 203Hg was studied. The content of 203Hg was lower in kidney and higher in liver and blood in the groups treated with selenate or selenite when compared with rats which received only mercury. The brain content of 203Hg was significantly increased in rats injected with selenite. Both selenium compounds injected immediately after mercury significantly decreased urinary as well as biliary excretion of 203Hg. A transient increase in the rate of biliary excretion of 203Hg during the first 2 h after administration was observed in rats treated with selenate. This finding seems to support the idea that the reduction of selenate to selenite in the body is not rapid but takes at least several hours.


The effect of oxygen tension on rat hepatocytes in short-term culture.
Cell viability, cytochrome P-450 content, cell respiration, and lipid peroxidation were all investigated as a function of oxygen tension in adult rat hepatocytes in short-term culture (less than 9 h). The various oxygen tensions used in this study were obtained by equilibrating culture medium with air, air + nitrogen, or air + oxygen. Cell viability, as assessed by trypan blue exclusion, was significantly greater at all time points tested when hepatocytes were cultured in Ham's F12 medium containing 132 microM O2, as compared to medium equilibrated with air (220 microM O2) or air + oxygen (298 microM O2). Cells cultured in 220 microM O2 (air) also exhibited a gradual loss of cytochrome P-450, so that by 9 h of incubation less than 60% of the active material remained. This loss of P-450 was minimized when cells were cultured in 163 microM O2 and abolished when cells were cultured in 132 microM O2. The 132 microM O2 exposure conditions also maintained cell respiration at the 1 h incubation values, whereas there was a continuous loss in cell respiration over time when the cells were cultured in either 220 microM O2 (air) or 298 microM O2 (air:O2). These cytotoxicity findings may be related to oxidative cell damage inasmuch as it was additionally demonstrated that lipid peroxidation (as measured by malondieldehyde equivalents) was consistantly lower in hepatocytes cultured in air:N2 as compared to air or air:O2. These results suggest that hepatocyte culture in low oxygen tension improves not only cell viability but also maintains other functional characteristics of the cell.


Splenic injury: nonoperative management in three patients with infectious mononucleosis.
Three patients with infectious mononucleosis and splenic injury were managed nonoperatively. Mechanisms of injury included significant left upper quadrant trauma during a football game in a 17-year-old boy, a minor fall in a 16-year-old girl, and no apparent trauma in the remaining patient, a 28-year-old man. All had significant abdominal pain and two were hypotensive upon admission. All had computerized tomographic scan documentation of splenic enlargement, significant injury, and free peritoneal blood. Paracentesis was not performed. Only the first patient received transfused blood. The hospitalization period ranged from nine to 14 days. Gradual and finally complete healing (return to normal size) was documented in all three patients. Follow-up is 66, 14, and 10 months. Although clearly controversial and not free from danger, successful nonoperative management of the injured enlarged spleen in select patients with mononucleosis is possible in an appropriate setting.


The current state, challenges, and future directions of deep brain stimulation for obsessive compulsive disorder.
Obsessive-compulsive disorder (OCD) is clinically and pathologically heterogenous, with symptoms often refractory to first-line treatments. Deep brain stimulation (DBS) for the treatment of refractory OCD provides an opportunity to adjust and individualize neuromodulation targeting aberrant circuitry underlying OCD. The tailoring of DBS therapy may allow precision in symptom control based on patient-specific pathology. Progress has been made in understanding the potential targets for DBS intervention; however, a consensus on an optimal target has not been agreed upon. A literature review of DBS for OCD was performed by querying the PubMed database. The following topics were covered: the evolution of DBS targeting in OCD, the concept of an underlying unified connectomic network, current DBS targets, challenges facing the field, and future directions which could advance personalized DBS in this challenging population. To continue the increasing efficacy of DBS for OCD, we must further explore the optimal DBS response across clinical profiles and neuropsychiatric domains of OCD as well as how interventions targeting multiple points in an aberrant circuit, multiple aberrant circuits, or a connectivity hub impact clinical response. Additionally, biomarkers would be invaluable in programming adjustments and creating a closed-loop paradigm to address symptom fluctuation in daily life.


Adherence to the European food safety authority's dietary recommendations and colorectal cancer risk.
The European Food Safety Authority (EFSA) recently published dietary guidelines for the intakes of carbohydrates, fiber, fats and water. We evaluated their role on the risk of a specific disease, known to be related to diet. We used data from an Italian case-control study including 1953 colorectal cancer (CRC) cases and 4154 controls. We developed a so-called EFSA index summing up 1 point for adherence to each EFSA guideline. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) of CRC and its subsites were derived from unconditional multiple logistic regression models, for both the index and its components. When each EFSA index component was analyzed separately, we found significant increased risks of CRC for non adherence to the guidelines on linoleic (OR=1.20, 95% CI, 1.07-1.36) and alpha-linolenic fatty acids (OR=1.19, 95% CI, 1.06-1.34). When all the guidelines were included in the same model, no significant association emerged. Compared with minimal adherence, the ORs of CRC for subsequent EFSA index scores were 1.03 (95% CI, 0.72-1.47), 1.05 (95% CI, 0.75-1.48), 1.04 (95% CI, 0.81-1.60), 0.99 (95% CI, 0.69-1.43), and 1.04 (95% CI, 0.67-1.61). No significant association emerged for colon and rectal cancer separately, and for males and females. Overall adherence to the EFSA dietary guidelines is not associated to colorectal, colon and rectal cancer risk in our population. Adherence to guidelines on linoleic and alpha-linolenic fatty acids may have a modest beneficial role on CRC risk.


Risk factors associated with mortality among patients who had candidemia in a university hospital.
Bloodstream infection due to Candida spp. is a primary cause of morbidity and mortality in tertiary hospitals. In this retrospective study, we included patients with a positive blood culture for Candida spp. after 48 h of hospitalization. A total of 335 patients who had candidemia were included in this study. Risk factors associated with mortality were hospitalization in internal medicine units and surgical clinics, age >60 years, mechanical ventilation, orotracheal intubation, hemodialysis, corticosteroids use, and C. parapsilosis infection. This study highlights the importance of health care related to invasive procedures and actions to improve patient immunity.


Facilitators and challenges to conducting interdisciplinary research.
Complex, interconnected issues challenge the United States health care system and the patients and families it serves. System fragmentation, limited resources, rigid disciplinary boundaries, institutional culture, ineffective communication, and uncertainty surrounding health policy legislation are contributing to suboptimal care delivery and patient outcomes. These problems are too complex to be solved by a single discipline. Interdisciplinary research affords the opportunity to examine and solve some of these problems from a more integrative perspective using innovative and rigorous methodological designs. In this paper, we explore lessons learned from exemplars funded by the Robert Wood Johnson Foundation's Interdisciplinary Nursing Quality Research Initiative. The discussion is framed using an adaptation of the Interdisciplinary Research Model to evaluate improvements in individual health outcomes, health systems, and health policy. Barriers and facilitators to designing, conducting, and translating interdisciplinary research are discussed. Implications for health system and policy changes, including the need to provide funding mechanisms to implement interdisciplinary processes in both research and clinical practice, are provided.


Fatal Erosion Atrial Septal Defect Device.
Percutaneus device closure of atrial septal defects offers an alternative to conventional surgical repair. While procedure techniques and devices have improved, severe complications remain that are neither completely understood nor entire predictable. We present a case of a 19-year-old female with left atrial erosion leading to lethal pericardial tamponade 1 month after ASD II closure using the Nit Occlude ASD-R® (NOASD-R, pfm Medical, Cologne, Germany). To our knowledge this is the first reported lethal complication related to this device.


Secular Trends in the Incidence of Dementia in a Multi-Ethnic Community.
Determination of secular trends in cognitive aging is important for prioritization of resources, services, and research in aging populations. Prior studies have identified declining dementia incidence associated with changes in cardiovascular risk factors and increased educational attainment. However, few studies have examined these factors in multi-ethnic cohorts. To identify secular trends in the incidence rate of dementia in an elderly population. Participants in this study were drawn from the Washington Heights-Inwood Columbia Aging Project, a multi-ethnic cohort study of northern Manhattan residents aged 65 years and older. Cox proportional hazards models were used to examine differences in the incidence of dementia in cohorts recruited in 1992 and 1999, with age at dementia or age at last follow-up visit as the "time-to-event" variable. Overall, there was a 41% reduction in the hazard ratio for dementia among participants in the 1999 cohort compared with those in the 1992 cohort, adjusting for age, sex, race, and baseline memory complaints (HR = 0.59). The reduction in incidence was greatest among non-Hispanic Whites and African-Americans and lowest among Hispanic participants (HRs = 0.60, 0.52 and 0.64, respectively), and was associated with increases in level of educational attainment, especially among African-Americans. Reduction in incidence of dementia was also greater among persons 75 years or older than among younger participants (HR = 0.52 versus HR = 0.69). Our results support previous findings that secular trends in dementia incidence are changing, including in aging minority populations.


Inhibition of prostaglandin E1-induced elevation of cytoplasmic free calcium ion by protein kinase C-activating phorbol esters and diacylglycerol in Swiss 3T3 fibroblasts.
In quiescent cultures of Swiss 3T3 cells, prostaglandin E1 (PGE1) known to elevate cAMP increased rapidly cytoplasmic free Ca2+ concentration ([Ca2+]i) as measured with the fluorescent Ca2+ indicator quin2. The primary source of the PGE1-induced elevation of [Ca2+]i was extracellular. Pretreatment of the cells with various doses of 12-O-tetradecanoylphorbol-13-acetate (TPA), a potent protein kinase C-activating phorbol ester, inhibited the PGE1-induced elevation of [Ca2+]i in a dose-dependent manner. Inversely, TPA enhanced slightly the PGE1-induced increase of cAMP. TPA alone did not affect the basal level of [Ca2+]i or cAMP in the absence of PGE1. The inhibitory action of TPA on the PGE1-induced elevation of [Ca2+]i was mimicked by other protein kinase C-activating agents such as phorbol 12,13-dibutyrate and 1-oleoyl-2-acetylglycerol. 4 alpha-Phorbol 12,13-didecanoate known to be inactive for protein kinase C was ineffective in this capacity. Prolonged treatment of the cells with phorbol 12,13-dibutyrate resulted in the down-regulation and disappearance of protein kinase C. In these protein kinase C-deficient cells, PGE1 still elevated [Ca2+]i to the same extent as that in the control cells, but TPA did not inhibit the PGE1-induced elevation of [Ca2+]i. These results strongly suggest that protein kinase C serves as an inhibitor for PGE1-induced Ca2+ influx in Swiss 3T3 cells.


Validation of a low-cost simulation strategy for burn escharotomy training.
Escharotomy is the primary effective intervention to relieve constriction and impending vascular compromise in deep, circumferential or near-circumferential burns of the extremities and trunk. Training on escharotomy indications, technique and pitfalls is essential, as escharotomy is both an infrequent and high-risk procedure in civilian and military medical environments, including low-resource settings. Therefore, we aimed to validate an educational strategy that combines video-based instruction with a low-cost, low-fidelity simulation model for teaching burn escharotomy. Pre-hospital and hospital-based medical personnel, with varying degrees of burn care-related experience, participated in a one-hour training session. The first part of the training consisted of video-based instruction that described the indications, preparation, steps, pitfalls and complications associated with escharotomy. The second part of the training consisted of a supervised, hands-on simulation with a previously described low-cost, low-fidelity escharotomy model. Participants were then offered two psychometrically validated instruments to assess their learning experience. 40 participants were grouped according to prior burn care and surgical experience: attending surgeons (6), surgery and emergency medicine residents and fellows (26), medical students (5), and pre-hospital personnel (3). On two psychometrically validated questionnaires, participants at both the attending and trainee levels overwhelmingly confirmed that our educational strategy met best educational practices on the criteria of active learning, collaboration, diverse ways of learning, and high expectations; they also highly rated their satisfaction with and self-confidence under this learning strategy. An educational strategy that combines video-based instruction and a low-cost, low-fidelity escharotomy simulation model was successfully demonstrated with participants across a broad range of prior burn care experience levels. This strategy is easily reproducible and broadly applicable to increase the knowledge and confidence of medical personnel before they are called to perform escharotomy. Important applications include resource-limited environments and deployed military settings.


Activation of AMPK and inactivation of Akt result in suppression of mTOR-mediated S6K1 and 4E-BP1 pathways leading to neuronal cell death in in vitro models of Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by loss of dopaminergic neurons. Dysregulation of mammalian target of rapamycin (mTOR) has been implicated in the pathogenesis of PD. However, the underlying mechanism is incompletely elucidated. Here, we show that PD mimetics (6-hydroxydopamine, N-methyl-4-phenylpyridine or rotenone) suppressed phosphorylation of mTOR, S6K1 and 4E-BP1, reduced cell viability, and activated caspase-3 and PARP in PC12 cells and primary neurons. Overexpression of wild-type mTOR or constitutively active S6K1, or downregulation of 4E-BP1 in PC12 cells partially prevented cell death in response to the PD toxins, revealing that mTOR-mediated S6K1 and 4E-BP1 pathways due to the PD toxins were inhibited, leading to neuronal cell death. Furthermore, we found that the inhibition of mTOR signaling contributing to neuronal cell death was attributed to suppression of Akt and activation of AMPK. This is supported by the findings that ectopic expression of constitutively active Akt or dominant negative AMPKα, or inhibition of AMPKα with compound C partially attenuated inhibition of phosphorylation of mTOR, S6K1 and 4E-BP1, activation of caspase-3, and neuronal cell death triggered by the PD toxins. The results indicate that PD stresses activate AMPK and inactivate Akt, causing neuronal cell death via inhibiting mTOR-mediated S6K1 and 4E-BP1 pathways. Our findings suggest that proper co-manipulation of AMPK/Akt/mTOR signaling may be a potential strategy for prevention and treatment of PD.


The effect of CO2 laser therapy on vaginal microcirculatory parameters in an animal model for genitourinary syndrome of menopause.
Vaginal laser therapy for the treatment of genitourinary syndrome of menopause (GSM) has been introduced to the market with limited (pre)clinical and experimental evidence supporting its efficacy. It is suggested that vaginal laser therapy increases epithelial thickness and improves vascularization, but the underlying biological working mechanism has not been substantiated yet. To evaluate the effects of CO2 laser therapy on vaginal atrophy using noninvasive incident dark field (IDF) imaging in a large animal model for GSM. An animal study was conducted between 2018 and 2019 and included 25 Dohne Merino ewes, of which 20 underwent bilateral ovariectomy (OVX) to induce iatrogenic menopause, and 5 did not. The total study duration was 10 months. Five months after OVX, ovariectomized ewes received monthly applications of CO2 laser (n = 7), vaginal estrogen (n = 7), or no treatment (n = 6) for 3 months. IDF imaging was performed monthly in all animals. The primary outcome was the proportion of image sequences containing capillary loops (angioarchitecture). Secondary outcomes included focal depth (epithelial thickness), and quantitative measures of vessel density and perfusion. Treatment effects were evaluated using ANCOVA and binary logistic regression. Compared to OVX-only, ewes treated with estrogen demonstrated a higher capillary loops proportion (4% vs. 75%, p < 0.01), and higher focal depth (60 (IQR 60-80) vs. 80 (IQR 80-80) p < 0.05). CO2 laser therapy did not change microcirculatory parameters. As the ewes' vaginal epithelium is thinner than that of humans, it may demand different laser settings. In a large animal model for GSM, CO2 laser therapy does not affect microcirculatory outcomes related to GSM, whereas vaginal estrogen treatment does. Until more homogeneous and objective evidence about its efficacy is available, CO2 laser therapy should not be adopted into widespread practice for treating GSM.


Curbing Obesity from One Generation to Another: the Effects of Bariatric Surgery on the In Utero Environment and Beyond.
Approximately 250,000 individuals seek bariatric surgery each year in the USA for the long-term resolution of obesity-related comorbidities. Greater than 80% of these individuals are women and approximately half are of child-bearing age. Although there are many positive metabolic benefits that are realized through surgical weight loss for both men and women, the various long-term hormonal, molecular, nutrient, and epigenetic changes following bariatric surgery have not been evaluated for the surgical recipient or in the context of pregnancy and the offspring. Pregnancy may be a vulnerable period of time for the bariatric surgery recipient, and thoughtful consideration of pregnancy management should be taken by health care providers and recipients alike. The purpose of this review is to explore potential etiologies of some of the gestation-specific outcomes for the mother and offspring.


Adsorption of ethinylestradiol (EE2) on polyamide 612: molecular modeling and effects of water chemistry.
This study demonstrates that ethinylestradiol (EE2), a priority estrogenic contaminant in water, can be rapidly and selectively removed from aqueous solutions using industrial-grade polyamide 612 (PA612) particles as adsorbents. Isothermal studies showed that nonporous low surface area (20 m(2) g(-1)) PA612 particles had a maximum adsorption capacity of 25.4 mg g(-1) for EE2 in water, which is higher or comparable to the results obtained with two benchmark activated carbon (AC) adsorbents (10.4-27.6 mg g(-1)). The adsorption of EE2 on PA612 followed pseudo-second order kinetics with a high adsorption rate exceeding those of the ACs by 5.3- to 22.4-fold. Computational chemistry calculations and molecular modeling showed that the strong binding affinity between EE2 and PA612 originates from the hydrophobic partitioning of EE2 solutes and hydrogen bonding interactions on PA612 amide groups. PA612 showed high adsorption selectivity for EE2 in water with highly consistent adsorption capacities for EE2 under the influence of a range of water chemistry parameters, including water salinity (NaCl, 1 mM-1 M), metal ions (K(I), Ca(II), and Zn(II); 0.1 M), natural organic matter (humic acids, 0.2-5 mg L(-1)), and pH level (4.8-9.1).


Impact of mode of conception on early pregnancy human chorionic gonadotropin rise and birth weight.
To assess whether the mode of conception and embryo biopsy impact first-trimester human chorionic gonadotropin (hCG) dynamics and subsequent risk of small for gestational age (SGA) or large for gestational age (LGA). Retrospective cohort study. University fertility center. Six hundred-two pregnant patients with singleton live births. Serial serum hCG measurements were obtained between 10 and 28 days postconception to determine the within-woman rate of change in hCG (slope) by mode of conception (unassisted pregnancy, fresh embryo transfer (ET), frozen ET, and frozen ET following preimplantation genetic testing for aneuploidy (PGT-A). Primary outcomes included birth weight, SGA, and LGA. Mode of conception is not independently associated with birth weight, SGA, or LGA. Mediation analysis revealed an expected one-day increase in log-transformed hCG varied by mode of conception: unassisted (0.41), fresh ET (0.39), frozen ET (0.42), PGT-A (0.44). Human chorionic gonadotropin rise has a positive effect on birth weight (55 g per SD increase in hCG slope) and is associated with SGA (odds ratio, 0.65), but not with LGA (odds ratio, 1.18). Human chorionic gonadotropin rise is an important mediator of the mode of conception/birth weight relationship. Preimplantation genetic testing for aneuploidy has the highest rate of hCG rise, followed by frozen ET, unassisted, and fresh ET. Faster rise is associated with higher birth weight and lower risk of SGA but does not impact LGA risk. Importantly, PGT-A does not increase the risk of extreme birth weight relative to other modes of conception evaluated.


Hearing Loss in Pediatrics: What the Medical Home Needs to Know.
Screening infants for hearing loss at birth is a standard in most states in the United States, but follow-up continues to warrant improvement. Understanding the definition of hearing loss, its etiology, appropriate intervention options, and knowledge of methods to optimize an infant's outcomes through the medical home can help to maximize speech and language skills.