The Kock pouch as a urinary reservoir. Pitfalls and perspectives.
The Kock continent urinary reservoir is a feasible alternative to traditional methods of urinary diversion in properly motivated patients who are anxious to exchange a convenient external appliance for frequent reservoir catheterization. The major shortcomings are increased operative time, loss of a long segment of small bowel, and an approximate 10 percent incidence of major reoperation. Long-term results are unknown. Facility with the operation requires extensive experience and attention to detail. Most complications are related to inadequate fixation of the intussuscepted valves and improper placement of a short terminal segment in the abdominal wall. Early repair of malfunctioning valves and distal segments that are difficult to catheterize is essential to prevent more severe acute complications. Whether the Kock pouch will replace the standard methods of urinary diversion must be determined by more extensive clinical experience and long-term results.


Progressive axonopathy when oligodendrocytes lack the myelin protein CMTM5.
Oligodendrocytes facilitate rapid impulse propagation along the axons they myelinate and support their long-term integrity. However, the functional relevance of many myelin proteins has remained unknown. Here, we find that expression of the tetraspan-transmembrane protein CMTM5 (chemokine-like factor-like MARVEL-transmembrane domain containing protein 5) is highly enriched in oligodendrocytes and central nervous system (CNS) myelin. Genetic disruption of the Cmtm5 gene in oligodendrocytes of mice does not impair the development or ultrastructure of CNS myelin. However, oligodendroglial Cmtm5 deficiency causes an early-onset progressive axonopathy, which we also observe in global and tamoxifen-induced oligodendroglial Cmtm5 mutants. Presence of the WldS mutation ameliorates the axonopathy, implying a Wallerian degeneration-like pathomechanism. These results indicate that CMTM5 is involved in the function of oligodendrocytes to maintain axonal integrity rather than myelin biogenesis.


Adaptive fuzzy output-feedback controller design for nonlinear systems via backstepping and small-gain approach.
This paper focuses on an input-to-state practical stability (ISpS) problem of nonlinear systems which possess unmodeled dynamics in the presence of unstructured uncertainties and dynamic disturbances. The dynamic disturbances depend on the states and the measured output of the system, and its assumption conditions are relaxed compared with the common restrictions. Based on an input-driven filter, fuzzy logic systems are directly used to approximate the unknown and desired control signals instead of the unknown nonlinear functions, and an integrated backstepping technique is used to design an adaptive output-feedback controller that ensures robustness with respect to unknown parameters and uncertain nonlinearities. This paper, by applying the ISpS theory and the generalized small-gain approach, shows that the proposed adaptive fuzzy controller guarantees the closed-loop system being semi-globally uniformly ultimately bounded. A main advantage of the proposed controller is that it contains only three adaptive parameters that need to be updated online, no matter how many states there are in the systems. Finally, the effectiveness of the proposed approach is illustrated by two simulation examples.


Numerical models, geochemistry and the zero-paradox noble-gas mantle.
Numerical models of whole-mantle convection demonstrate that degassing of the mantle is an inefficient process, resulting in ca. 50% of the (40)Ar being degassed from the mantle system. In this sense the numerical simulations are consistent with the (40)Ar mass balance between the atmosphere and mantle reservoir. These models, however, are unable to preserve the large-scale heterogeneity predicted by models invoking geochemical layering of the mantle system. We show that the three most important noble-gas constraints on the geochemically layered mantle are entirely dependent on the (3)He concentration of the convecting mantle derived from the (3)He flux into the oceans and the average ocean-crust generation rate. A factor of 3.5 increase in the convecting-mantle noble-gas concentration removes all requirements for: a (3)He flux into the upper mantle from a deeper high (3)He source; a boundary in the mantle capable of separating heat from helium; and a substantial deep-mantle reservoir to contain a hidden (40)Ar rich reservoir. We call this model concentration for the convecting mantle the 'zero-paradox' concentration. The time-integrated flux of (3)He into the oceans is a robust observation, but only representative of the ocean-floor activity over the last 1000 years. In contrast, ocean-floor generation occurs over tens of millions of years. We argue that combining these two observations to obtain the (3)He concentration of the mantle beneath mid-ocean ridges is unsound. Other indicators of mantle (3)He concentration suggest that the real value may be at least a factor of two higher. As the zero-paradox concentration is approached, the noble-gas requirement for mantle layering is removed. We further consider the role that recycled material plays in ocean-island-basalt generation and show that a source with high (3)He and (3)He/(4)He must exist within the mantle. Nevertheless, only a small amount of this material is required to generate both the observed ocean-island (3)He/(4)He ratios and the concentrations inferred from basalt samples for this mantle source.


[Effect of vasodilator preparations on the ATPase activity of the erythrocyte membranes of rabbits with experimental vascular pathology].
Experiments on rabbits with pituitrin hypertension have demonstrated activation of Na+, K+-ATPase of red cell membranes. Meanwhile ATPase activity declined in vasorenal hypertension. The vasodilatory drugs (papaverine, pentamine, euphylline, nicotinic acid) administered in courses normalized both the activity of transport ATPases of cell membranes and carbohydrate metabolism of blood vessels, which was disturbed in experimental vascular pathology.


Pyrolysis mass spectrometry in epidemiological and population genetic studies of Haemophilus influenzae.
Haemophilus influenzae serotype b (Hib) vaccines have reduced the amount of invasive Hib disease in immunised infants. However, Hib disease remains in unvaccinated infants and adults and non-capsulate H. influenzae (NCHi) still causes infections, including outbreaks of respiratory disease. Characterisation of strains and the bacterial population as a whole is therefore necessary to detect outbreaks of infection with NCHi or changes in the population, for example, to vaccine-resistant clones of Hib. The rapid, simple and objective technique of pyrolysis mass spectrometry (PMS) was investigated as an alternative to current complex, subjective methods. PMS was compared with ribotyping and multilocus enzyme electrophoresis (MLEE) for population genetic analyses of Hib and with ribotyping and protein profiling for epidemiological analyses of NCHi. PMS clustered all the isolates of Hib together whereas MLEE and ribotyping distinguished certain clones - this is probably because the three methods examine different (and unrelated) characteristics of the organisms. The PMS results were essentially similar to those from ribotyping and protein profiling for the epidemiological analyses of outbreaks of NCHi disease. Therefore, PMS is probably unsuitable for comparisons of Hib populations but it is a useful addition to the arsenal of techniques for the characterisation of NCHi.


Telomerase activity with concurrent loss of cell cycle regulation in feline post-traumatic ocular sarcomas.
Paraffin wax-embedded ocular globes of cats with post-traumatic ocular sarcomas were examined for the presence of TERT, the active subunit of telomerase. The latter is a ribonucleoprotein complex essential for immortalization and expressed by most malignant tumours, germ line cells, lens epithelial cells, and some stem cells. Due to the frequent loss of cell cycle control with the increased expression of telomerase activity, post-traumatic ocular sarcomas were also examined for loss of p16 expression and alterations in p53, the findings being related to mitotic score, tumour grade, and proliferating cell nuclear antigen. These sarcomas expressed telomerase at a high frequency (62.5%); in addition, the majority showed alterations in cell cycle control, as evaluated by lack of p16 immunolabelling (66.7%). Alterations in p53 were the sole mechanism by which cell cycle control was dysregulated in only two tumours expressing TERT (13%). These findings suggest that p16, and not p53, represents the primary mechanism by which post-traumatic ocular sarcomas that express telomerase activity escape cell cycle control.


Changes in myocardial fluid filtration are reflected in epicardial lymph pressure.
The effect of increased fluid filtration on stopped-flow epicardial lymph pressure (P(lymph)), used as an indicator of myocardial interstitial volume, was investigated in the anesthetized open-chest dog. Histamine infusion resulted in an increased systolic peak in the P(lymph) signal together with an increase in diastolic P(lymph) in four of five animals. During reactive hyperemia, systolic and diastolic P(lymph) increased to 127 +/- 8 and 121 +/- 6.7% (mean +/- SE, n = 6) of control, respectively. Peak P(lymph) was approximately 15 s later than peak coronary blood flow and venous pressure (P(ven)). When P(ven) was transiently elevated to 367 +/- 72 (systolic) and 247 +/- 45% (diastolic) of control, P(lymph) increased to 132 +/- 12 and 120 +/- 5.5% of control. The time of response was similar for P(ven) and P(lymph) (t50 approximately 2 S). The increased systolic and diastolic P(lymph) can be explained by an increase in interstitial and lymph filling. It is concluded that changes in myocardial fluid filtration are reflected in epicardial P(lymph). Furthermore, it seems that cardiac contraction constitutes an important defense mechanism against the formation of myocardial edema.


Medicalized menopause, US style.
The construction of menopause as an estrogen deficiency condition originated primarily in the United States and is being exported to many other countries throughout the world. I have identified specific cultural assumptions and policies in the United States as well as the ubiquitous gender biases that have contributed to the medicalization of menopause. Since medicalized menopause is causing harm to women worldwide, it is imperative that it be addressed by the international community.


Late arthroscopic retrieval of a bullet from hip joint.
We describe a case of arthroscopic retrieval of a bullet from the hip joint of an 18-year-old boy, who sustained the injury four months back, accidentally, while bird hunting with a country made shotgun. The surgery was performed with the standard ordinary instrumentation of knee arthroscopy. The patient became pain-free the same evening and started partial weight bearing on the next day of surgery. At 13 months follow-up, the patient had returned to normal activity without any functional limitations.


Immune reconstitution/immunocompetence in recipients of kidney plus hematopoietic stem/facilitating cell transplants.
Nineteen subjects have more than 18 months' follow-up in a phase IIb tolerance protocol in HLA-mismatched recipients of living donor kidney plus facilitating cell enriched hematopoietic stem cell allografts (FCRx). Reduced intensity conditioning preceded a kidney allograft, followed the next day by FCRx. Twelve have achieved stable donor chimerism and have been successfully taken off immunosuppression (IS). We prospectively evaluated immune reconstitution and immunocompetence. Return of CD4 and CD8 T central and effector memory cell populations was rapid. T-cell receptor (TCR) Excision Circle analysis showed a significant proportion of chimeric cells produced were being produced de novo. The TCR repertoires posttransplant in chimeric subjects were nearly as diverse as pretransplant donors and recipients, and were comparable to subjects with transient chimerism who underwent autologous reconstitution. Subjects with persistent chimerism developed few serious infections when off IS. The majority of infectious complications occurred while subjects were still on conventional IS. BK viruria and viremia resolved after cessation of IS and no tissue-invasive cytomegalovirus infections occurred. Notably, although 2 of 4 transiently or nonchimeric subjects experienced recurrence of their underlying autoimmune disorders, none of the chimeric subjects have, suggesting that self-tolerance is induced in addition to tolerance to alloantigen. No persistently chimeric subject has developed donor-specific antibody, and renal function has remained within normal limits. Patients were successfully vaccinated per The American Society for Blood and Marrow Transplantation guidelines without loss of chimerism or rejection. Memory for hepatitis vaccination persisted after transplantation. Chimeric subjects generated immune responses to pneumococcal vaccine. These data suggest that immune reconstitution and immunocompetence are maintained in persistently chimeric subjects.


Self-assembly of Janus Au:Fe3O4 branched nanoparticles. From organized clusters to stimuli-responsive nanogel suprastructures.
Janus nanoparticles offer enormous possibilities through a binary selective functionalization and dual properties. Their self-assembly has attracted strong interest due to their potential as building blocks to obtain molecular colloids, supracrystals and well-organized nanostructures that can lead to new functionalities. However, this self-assembly has been focused on relatively simple symmetrical morphologies, while for complex nanostructures this process has been unexplored. Here, we study the assembly of plasmonic-magnetic Janus nanoparticles with a branched (nanostar) - sphere morphology. The branched morphology enhances their plasmonic properties in the near-infrared region and therefore their applicability, but at the same time constrains their self-assembly capabilities to obtain more organized or functional suprastructures. We describe the self-assembly of these nanoparticles after amphiphilic functionalization. The role of the nanoparticle branching, as well as the size of the polymer-coating, is explored. We show how the use of large molecular weight stabilizing polymers can overcome the anisotropy of the nanoparticles producing a change in the morphology from small clusters to larger quasi-cylindrical nanostructures. Finally, the Janus nanoparticles are functionalized with a thermo-responsive elastin-like recombinamer. These nanoparticles undergo reversible self-assembly in the presence of free polymer giving rise to nanoparticle-stabilized nanogel-like structures with controlled size, providing the possibility to expand their applicability to multi-stimuli controlled self-assembly.


[Study on separation and determination of four tropane alkaloids in crude drugs by micellar HPLC].
A new method for separation and determination of four important tropane alkaloids (hyoscyamine, scopolamine, anisodamine and anisodine) in crude drugs by micellar HPLC was developed. The mobile phase was optimized with the modified simplex method (MSM). The chromatographic response function (CRF) was used as the criterion of optimization and three dimension simplex was used in this course. After eight tests, the given accuracy was achieved. In the optimized system the four tropane alkaloids are separated not only from each other but also from the interfering components in the crude drugs. The CMC of SDS in mobile phase was determined to be 5 mmol/L by circle method. The experimental evidences show that the optimized system is micellar chromatography. This method is simple, sensitive and accurate. The extraction solution can be injected directly without the need of a general purification procedure. Several plant materials containing tropane alkaloids were determined. The results and chromatograms are reported in this paper.


Wnt/β-Catenin Regulates the Activity of Epiprofin/Sp6, SHH, FGF, and BMP to Coordinate the Stages of Odontogenesis.
We used an in vitro tooth development model to investigate the effects of overactivation of the Wnt/β-catenin pathway during odontogenesis by bromoindirubin oxime reagent (BIO), a specific inhibitor of GSK-3 activity. Overactivating the Wnt/β-catenin pathway at tooth initiation upregulated and ectopically expressed the epithelial markers Sonic Hedgehog (Shh), Epiprofin (Epfn), and Fibroblast growth factor8 (Fgf8), which are involved in the delimitation of odontogenic fields in the oral ectoderm. This result indicated an ectopic extension of the odontogenic potential. During tooth morphogenesis, Fibroblast growth factor4 (Fgf4), Fibroblast growth factor10 (Fgf10), Muscle segment homeobox 1 (Msx-1), Bone Morphogenetic protein 4 (Bmp4), and Dickkopf WNT signaling pathway inhibitor 1 (Dkk-1) were overexpressed in first molars cultured with BIO. Conversely, the expression levels of Wingless integration site 10b (Wnt-10b) and Shh were reduced. Additionally, the odontoblast differentiation markers Nestin and Epfn showed ectopic overexpression in the dental mesenchyme of BIO-treated molars. Moreover, alkaline phosphatase activity increased in the dental mesenchyme, again suggesting aberrant, ectopic mesenchymal cell differentiation. Finally, Bmp4 downregulated Epfn expression during dental morphogenesis. We suggest the presence of a positive feedback loop wherein Epfn and β-catenin activate each other. The balance of the expression of these two molecules is essential for proper tooth development. We propose a possible link between Wnt, Bmp, and Epfn that would critically determine the correct patterning of dental cusps and the differentiation of odontoblasts and ameloblasts.


A novel graph-based k-partitioning approach improves the detection of gene-gene correlations by single-cell RNA sequencing.
Gene expression is regulated by transcription factors, cofactors, and epigenetic mechanisms. Coexpressed genes indicate similar functional categories and gene networks. Detecting gene-gene coexpression is important for understanding the underlying mechanisms of cellular function and human diseases. A common practice of identifying coexpressed genes is to test the correlation of expression in a set of genes. In single-cell RNA-seq data, an important challenge is the abundance of zero values, so-called "dropout", which results in biased estimation of gene-gene correlations for downstream analyses. In recent years, efforts have been made to recover coexpressed genes in scRNA-seq data. Here, our goal is to detect coexpressed gene pairs to reduce the "dropout" effect in scRNA-seq data using a novel graph-based k-partitioning method by merging transcriptomically similar cells. We observed that the number of zero values was reduced among the merged transcriptomically similar cell clusters. Motivated by this observation, we leveraged a graph-based algorithm and develop an R package, scCorr, to recover the missing gene-gene correlation in scRNA-seq data that enables the reliable acquisition of cluster-based gene-gene correlations in three independent scRNA-seq datasets. The graphically partitioned cell clusters did not change the local cell community. For example, in scRNA-seq data from peripheral blood mononuclear cells (PBMCs), the gene-gene correlation estimated by scCorr outperformed the correlation estimated by the nonclustering method. Among 85 correlated gene pairs in a set of 100 clusters, scCorr detected 71 gene pairs, while the nonclustering method detected only 4 pairs of a dataset from PBMCs. The performance of scCorr was comparable to those of three previously published methods. As an example of downstream analysis using scCorr, we show that scCorr accurately identified a known cell type (i.e., CD4+ T cells) in PBMCs with a receiver operating characteristic area under the curve of 0.96. Our results demonstrate that scCorr is a robust and reliable graph-based method for identifying correlated gene pairs, which is fundamental to network construction, gene-gene interaction, and cellular omic analyses. scCorr can be quickly and easily implemented to minimize zero values in scRNA-seq analysis and is freely available at https://github.com/CBIIT-CGBB/scCorr .


What do family caregivers know about palliative care? Results from a national survey.
Despite their key role in caring for individuals with serious, chronic illness, there have been no national studies examining family caregiver awareness and perceptions of palliative care. Hence, our objectives were to ascertain level of knowledge of palliative care among U.S. family caregivers and describe demographic variation in awareness and perceptions of palliative care. Using the 2018 National Cancer Institute Health Information National Trends Survey, we identified unpaid family caregivers caring or making healthcare decisions for someone with a medical, behavioral, disability, or other condition. Respondents were asked about their awareness of the term "palliative care" and, if aware, how much they agreed with statements representing common (mis)perceptions about palliative care (e.g., "Palliative care is the same as hospice"). More than one-half of caregivers (55%) had "never heard" of palliative care; 19.2% knew what palliative care was and "could explain it to someone else." In adjusted models, racial minorities (vs. whites) and those without a college degree were less likely to have heard of palliative care. Among those aware of palliative care, ~40% "strongly" or "somewhat" agreed that "Palliative care is the same as hospice"; another 10.5% "didn't know." Similarly, 40% reported that "When I think of palliative care, I automatically think of death." One-half of family caregivers of adults with serious chronic illness have never heard of palliative care. Even among those who had heard of palliative care, the majority do not distinguish it from hospice care and death. Given the role family caregivers may play in decisions to access palliative care, public messaging efforts are needed to clarify palliative care services in a way that is patient- and family-centered.


Multi-parametric MRI (mpMRI) for treatment response assessment of radiation therapy.
Magnetic resonance imaging (MRI) plays an important role in the modern radiation therapy (RT) workflow. In comparison with computed tomography (CT) imaging, which is the dominant imaging modality in RT, MRI possesses excellent soft-tissue contrast for radiographic evaluation. Based on quantitative models, MRI can be used to assess tissue functional and physiological information. With the developments of scanner design, acquisition strategy, advanced data analysis, and modeling, multiparametric MRI (mpMRI), a combination of morphologic and functional imaging modalities, has been increasingly adopted for disease detection, localization, and characterization. Integration of mpMRI techniques into RT enriches the opportunities to individualize RT. In particular, RT response assessment using mpMRI allows for accurate characterization of both tissue anatomical and biochemical changes to support decision-making in monotherapy of radiation treatment and/or systematic cancer management. In recent years, accumulating evidence have, indeed, demonstrated the potentials of mpMRI in RT response assessment regarding patient stratification, trial benchmarking, early treatment intervention, and outcome modeling. Clinical application of mpMRI for treatment response assessment in routine radiation oncology workflow, however, is more complex than implementing an additional imaging protocol; mpMRI requires additional focus on optimal study design, practice standardization, and unified statistical reporting strategy to realize its full potential in the context of RT. In this article, the mpMRI theories, including image mechanism, protocol design, and data analysis, will be reviewed with a focus on the radiation oncology field. Representative works will be discussed to demonstrate how mpMRI can be used for RT response assessment. Additionally, issues and limits of current works, as well as challenges and potential future research directions, will also be discussed.


Prevalence of Hemoproteus iwa in Galapagos great frigatebirds (Fregata minor) and their obligate fly ectoparasite (Olfersia spinifera).
The prevalence of hemosporidian parasites varies among different host species, geographic locations, habitats, and host life histories, and yet we do not have a firm understanding of the ultimate causes of the variation. Seabirds are not typically found infected with hemosporidian parasites; however, frigatebird species have been repeatedly documented with Hemoproteus spp. infections. Hemoproteus iwa in Galapagos great frigatebirds (Fregata minor) is vectored by a hippoboscid fly, Olfersia spinifera , an obligate ectoparasite of the bird host. Five populations of Galapagos great frigatebirds, and flies collected from the birds, were sampled and tested for H. iwa . Prevalence did not differ across 4 yr or between 5 islands, but males were found to have a significantly higher prevalence of infection than did females. Additionally, juveniles were more likely to be infected than were adults and chicks. Because the invertebrate vector is an obligate parasite, we were able to estimate prevalence in the vector as well as in the particular host upon which it fed, a task that is impossible, or nearly impossible, in hemosporidian parasites vectored by midges or mosquitoes. We tested the correlation between the infection status of the bird host and the infection status of the fly collected from the bird. More often than not the 2 were correlated, but some mismatches were found. Using the occurrence of infected flies on uninfected birds (12/99) as a proxy for transmission potential, we can estimate the transmission rate to be between 5 and 20% (95% confidence intervals) among individual vertebrate hosts.


Use of the peripheral nerve stimulator and standard, unsheathed needles in performing regional nerve blocks.
Although the use of peripheral nerve stimulators and unsheathed needles in performing nerve blocks has been previously described, there has been a growing emphasis on the use of specific, expensive equipment to ensure success. The availability of equipment designed for use in peripheral nerve blocks, insulated needles and nerve stimulators, will help promote the use of these techniques, but the expense of purchasing this specialized equipment may contribute to a decline in performance of certain types of blocks. This article reviews the use of standard equipment, available in most anesthesia departments, for performing peripheral nerve blocks successfully.


Viral proliferation and expression of tumor-related gene in different chicken embryo fibroblasts infected with different tumorigenic phenotypes of avian leukosis virus subgroup J.
Subgroup J avian leukosis virus (ALV-J) causes a neoplastic disease in infected chickens. The ALV-J strain NX0101, which was isolated from broiler breeders in 2001, mainly induced formation of myeloid cell tumors. However, strain HN10PY01, which was recently isolated from laying hens, mainly induces formation of myeloid cell tumors and hemangioma. To identify the molecular pathological mechanism underlying changes in host susceptibility and tumor classification induced by these two types of ALV-J strains, chicken embryo fibroblasts derived from chickens with different genetic backgrounds (broiler breeders and laying hens) and an immortalized chicken embryo fibroblasts (DF-1) were prepared and infected with strain NX0101 or HN10PY01, respectively. The 50% tissue culture infective dose (TCID50) and levels of ALV group-specific antigen p27 and heat shock protein 70 in the supernatant collected from the ALV-J infected cells were detected. Moreover, mRNA expression levels of tumor-related genes p53, c-myc, and Bcl-2 in ALV-J-infected cells were quantified. The results indicated that the infection of ALV-J could significantly increase mRNA expression levels of p53, c-myc, and Bcl-2 Strain HN10PY01 exhibited a greater influence on the three tumor-related genes in each of the three types of cells when compared with strain NX0101, and the TCID50 and p27 levels in the supernatant collected from HN10PY01-infected cells were higher than those collected from NX0101-infected cells. These results indicate that the infection of the two ALV-J strains influenced the gene expression levels in the infected cells, while the newly isolated strain HN10PY01 showed higher replication ability in cells and induced higher expression levels of tumor-related genes in infected cells. Furthermore, virus titers and expression levels of tumor-related genes and cellular stress responses of cells with different genetic backgrounds when infected with each of the two ALV-J strain were different, indicating that genetic backgrounds influenced the capabilities of the virus to infect and proliferate. The findings of this study provide useful data to further elucidate the mechanism underlying host susceptibility and tumor classification in ALV-J-infected chickens and cells.


A neutral polysaccharide with a triple helix structure from ginger: Characterization and immunomodulatory activity.
A neutral ginger polysaccharide fraction (NGP) was isolated from ginger, and its primary structures and immunomodulatory activity were investigated. The results showed that NGP had a low molecular weight of 6305 Da. NGP principally consisted of glucose (93.88%) together with minor levels of galactose (3.27%) and arabinose (1.67%). Besides, results of methylation analysis and 1D/2D NMR spectroscopy demonstrated that NGP was α-glucan which had the main chain of 1,4-linked α-d-Glcp and α-d-Glcp residues branched at C-6 position which was different from the common triple helical β-glucans. NGP also displayed a remarkable immunological activity on the RAW264.7 cells in vitro. It could significantly enhance the proliferation of macrophages without cytotoxicity and increase the production of immune substances (NO, TNF-α, IL-1β and IL-6). The secretion at the concentration of 200 μg/mL was 29.41 μM, 1496.71, 44.30 and 1889.83 pg/mL for each substance, respectively. The results indicated that NGP could be a potential immune agent and might provide meaningful information for further chain conformation and immune mechanism research.


Baseline multimodal information predicts future motor impairment in premanifest Huntington's disease.
In Huntington's disease (HD), accurate estimates of expected future motor impairments are key for clinical trials. Individual prognosis is only partially explained by genetics. However, studies so far have focused on predicting the time to clinical diagnosis based on fixed impairment levels, as opposed to predicting impairment in time windows comparable to the duration of a clinical trial. Here we evaluate an approach to both detect atrophy patterns associated with early degeneration and provide a prognosis of motor impairment within 3 years, using data from the TRACK-HD study on 80 premanifest HD (pre-HD) individuals and 85 age- and sex-matched healthy controls. We integrate anatomical MRI information from gray matter concentrations (estimated via voxel-based morphometry) together with baseline data from demographic, genetic and motor domains to distinguish individuals at high risk of developing pronounced future motor impairment from those at low risk. We evaluate the ability of models to distinguish between these two groups solely using baseline imaging data, as well as in combination with longitudinal imaging or non-imaging data. Our models show improved performance for motor prognosis through the incorporation of imaging features to non-imaging data, reaching 88% cross-validated accuracy when using baseline non-longitudinal information, and detect informative correlates in the caudate nucleus and the thalamus both for motor prognosis and early atrophy detection. These results show the plausibility of using baseline imaging and basic demographic/genetic measures for early detection of individuals at high risk of severe future motor impairment in relatively short timeframes.


DNA breakdown by the 4-quinolones and its significance.
DNA breakdown occurred in Escherichia coli KL16 exposed to nalidixic acid, ciprofloxacin or norfloxacin. However DNA breakdown does not seem to be the cause of the lethality of the 4-quinolones because it still occurred under conditions which abolished the lethality of nalidixic acid. Furthermore, no correlation was found between the amount of DNA breakdown and the rate of death of bacteria caused by the three 4-quinolones. Similarly, DNA breakdown did not occur when recB or recC mutants were treated with nalidixic acid despite both mutants being killed by the drug, again suggesting that DNA breakdown is not the cause of bacterial death. Since recB and recC mutants lack exonuclease V, this enzyme may be responsible for the DNA breakdown observed in bacteria treated with 4-quinolones.


Changes in microsomal phospholipases and arachidonic acid in experimental alcoholic liver injury: relationship to cytochrome P-450 2E1 induction and conjugated diene formation.
We evaluated the role of changes in microsomal phospholipases (A and C) and arachidonic acid in the intragastric rat feeding model. The experimental animals (male Wistar rats), divided into 4-5 rats/group, were fed the following diets: corn oil and ethanol and corn oil plus dextrose. One set of groups was killed after 2 weeks of feeding, and the second set was killed after 1 month. For each animal, microsomal analysis of cytochrome P-450 2E1 (CYP 2E1) and fatty acids was done. Fourteen animals had analyses of phospholipase C (PLC) and phospholipase A (PLA), and 10 animals had measurements of conjugated dienes. A significant correlation was obtained between the level of CYP 2E1 and the decrease in arachidonic acid (AA) from baseline levels (r = 0.69, p < 0.01). The decrease in AA also correlated with the increase in conjugated dienes (r = 0.70, p < 0.05). PLA and PLC activities were both significantly increased in the corn oil and ethanol groups. The activity of PLC correlated with the decline in AA (r = 0.69, p < 0.01). The correlations noted between the decrease in microsomal AA and CYP 2E1 induction and conjugated diene formation suggest that these processes may be interlinked especially in regard to generation of lipid peroxides that may play a role in alcoholic liver injury.


Coexisting disorders and problems in preschool children with autism spectrum disorders.
To analyze cooccurring disorders and problems in a representative group of 198 preschool children with autism spectrum disorders (ASD) who had had interventions at a specialized habilitation center. Parents and children were seen by a research team. Data were based on parental interviews, pediatric assessments, and tests of the child. Information on autistic symptoms, general cognitive function, speech and language, motor function, epilepsy, vision, hearing, activity level, behavior, and sleep was collected. Three ASD categories were used: (1) autistic disorder (AD), (2) autistic-like condition (ALC) or Asperger syndrome, and (3) one group with autistic symptoms/traits but not entirely all its criteria met for ASD. Children with autism had a mean of 3.2 coexisting disorders or problems, the ALC/Asperger group had a mean of 1.6, and children with autistic traits had a mean of 1.6. The most common disorder/problems in the total group pertained to language problems (78%), intellectual disability (ID) (49%), below average motor function (37%), and severe hyperactivity/ADHD (33%). The results accord with the concept of early symptomatic syndromes eliciting neurodevelopmental clinical examination (ESSENCE), and highlight the need of considering ASD in a broad perspective taking also other cooccurring developmental disorders into account.


Re-evaluation of acetic acid, lactic acid, citric acid, tartaric acid, mono- and diacetyltartaric acid, mixed acetic and tartaric acid esters of mono- and diglycerides of fatty acids (E 472a-f) as food additives.
The Panel on Food Additives and Flavourings (FAF) provided a scientific opinion re-evaluating the safety of acetic acid, lactic acid, citric acid, tartaric acid, mono- and diacetyltartaric acids, mixed acetic and tartaric acid esters of mono- and diglycerides of fatty acids (E 472a-f) as food additives. All substances had been previously evaluated by the Scientific Committee for Food (SCF) and by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Hydrolysis of E472a,b,c,e was demonstrated in various experimental systems, although the available data on absorption, distribution, metabolism, excretion (ADME) were limited. The Panel assumed that E472a-f are extensively hydrolysed in the GI tract and/or (pre-)systemically after absorption into their individual hydrolysis products which are all normal dietary constituents and are metabolised or excreted intact. No adverse effects relevant for humans have been identified from the toxicological database available for E472a-f. The Panel considered that there is no need for a numerical acceptable daily intake (ADI) for E 472a,b,c. The Panel also considered that only l(+)-tartaric acid has to be used in the manufacturing process of E472d,e,f. The Panel established ADIs for E 472d,e,f based on the group ADI of 240 mg/kg body weight (bw) per day, expressed as tartaric acid, for l(+)-tartaric acid-tartrates (E334-337, 354) and considering the total amount of l(+)-tartaric acid in each food additive. Exposure estimates were calculated for all food additives individually, except for E 472e and f, using maximum level, refined exposure and food supplements consumers only scenarios. Considering the exposure estimates, there is no safety concern at their reported uses and use levels. In addition, exposure to tartaric acid released from the use of E 472d,e,f was calculated. The Panel also proposed a number of recommendations.


Immune-checkpoint inhibitors to treat cancers in specific immunocompromised populations: a critical review.
Because of their efficacy against numerous cancers, immune-checkpoint inhibitors (ICIs), anti-cytotoxic T-lymphocyte antigen-4, and anti-programmed cell death monoclonal antibodies are being used ever more often in oncology. However, some patients were excluded from clinical trials because of their comorbidities despite their potentially higher cancer frequencies, as is the case for immunocompromised patients. Areas covered: We analyzed reported preclinical and clinical information and evaluated the risk/benefit ratio for four immunocompromised populations: people living with human immunodeficiency virus (PLHs), solid-organ transplant recipients, recipients of hematopoietic stem-cell allografts, and patients with autoimmune diseases. Expert commentary: Information available in the literature is fragmentary and scarce, making it difficult to evaluate the risk/benefit ratio. It can, nonetheless, be noted that ICI use in PLHs seems possible. For solid-organ transplant recipients, the risk for the graft seems elevated. For the other two populations, it is difficult to conclude at this time.


Overproduction of the multidrug efflux pump MexEF-OprN does not impair Pseudomonas aeruginosa fitness in competition tests, but produces specific changes in bacterial regulatory networks.
It is generally assumed that acquisition of antibiotic resistance leads to non-specific fitness costs. We have tested the alternative hypothesis that acquisition of antibiotic resistance may not always produce a general burden to the microorganisms, as measured in competition tests, but rather lead to specific changes in bacterial physiology. To this end we studied the effect of overproducing the multidrug efflux pump MexEF-OprN on Pseudomonas aeruginosa due to a constitutive activation of MexT, the transcriptional activator of the mexEF-oprN genes. We found that overexpression of MexEF-OprN does not cause a significant decrease in P.aeruginosa fitness in classical competition tests, indicating the absence of a large metabolic burden and that any possible negative effects might be observed only under specific conditions. Transcriptomic analyses revealed that overexpression of MexEF-OprN results in reduced expression of several quorum-sensing regulated genes. We traced back this phenotype to a delay in PQS production due to extrusion of kynurenine, a PQS precursor, through the efflux pump. Type VI secretion was also impaired. A Caenorhabditis elegans model demonstrated that overproduction of MexEF-OprN impairs virulence in P.aeruginosa. This effect was mainly due to the activity of the efflux pump, and not to MexT, despite the fact that the latter regulates Type III and Type VI secretion. Altogether, these data indicate that antibiotic resistance can produce modifications in the bacterial regulatory networks with relevant consequences for the bacterial behaviour in specific ecosystems, including the infected host.


Recapitulation and Modulation of the Cellular Architecture of a User-Chosen Cell of Interest Using Cell-Derived, Biomimetic Patterning.
Heterogeneity of cell populations can confound population-averaged measurements and obscure important findings or foster inaccurate conclusions. The ability to generate a homogeneous cell population, at least with respect to a chosen trait, could significantly aid basic biological research and development of high-throughput assays. Accordingly, we developed a high-resolution, image-based patterning strategy to produce arrays of single-cell patterns derived from the morphology or adhesion site arrangement of user-chosen cells of interest (COIs). Cells cultured on both cell-derived patterns displayed a cellular architecture defined by their morphology, adhesive state, cytoskeletal organization, and nuclear properties that quantitatively recapitulated the COIs that defined the patterns. Furthermore, slight modifications to pattern design allowed for suppression of specific actin stress fibers and direct modulation of adhesion site dynamics. This approach to patterning provides a strategy to produce a more homogeneous cell population, decouple the influences of cytoskeletal structure, adhesion dynamics, and intracellular tension on mechanotransduction-mediated processes, and a platform for high-throughput cellular assays.


Cranial irradiation in adults diagnosed with acute myelogenous leukemia presenting with hyperleukocytosis and neurologic dysfunction.
This study describes our institution's experience using whole brain radiation therapy (WBRT) to treat patients with acute myelogenous leukemia (AML) presenting with hyperleukocytosis. After approval by the institutional review board, we identified patients with AML and hyperleukocytosis using hospital records. The primary endpoints in the study included alleviation of neurological symptoms (or prevention if prophylactic RT was used), overall survival, development of intracranial hemorrhage (ICH) and ≥ grade 3 toxicities using the Common Terminology Criteria for Adverse Events version 4.0 (CTCAE v4.0). Eighteen patients received WBRT for the treatment of AML hyperleukocytosis. Thirteen patients received treatment in order to control neurological symptoms. Clinical assessment showed that 12 of 13 patients (92%) achieved resolution of neurological symptoms either concurrent with RT or immediately after RT. The mean overall survival for all of the patients who received WBRT was 14.2 months (95% confidence interval, 5.4-23.0). No patient who received RT experienced ≥ grade 3 toxicity. Two (6%) patients developed ICH following therapy. Our institution's experience demonstrates that WBRT may be utilized as part of multimodality therapy in order to alleviate or prevent neurological symptoms in patients with AML presenting with leukostasis.


The Adolescent Knee Pain (AK-Pain) prognostic tool: protocol for a prospective cohort study.
Background: One in three children and adolescents experience knee pain. Approximately one in two adolescents with knee pain will continue to experience pain even five years later and have low quality of life. The general practitioner (GP) is the first point of contact for children and adolescents with knee pain in Denmark. There is a variety of treatments being delivered in general practice, despite similar symptoms and patients' characteristics. This suggests a need to support the GPs in identifying those at high risk of a poor outcome early on, in order to better allocate resources. The aim of this study is to develop a user-friendly prognostic tool to support GPs' management of children and adolescents' knee pain. Methods: A preliminary set of items in the prognostic tool were identified using systematic reviews and meta-analysis of individual participant data. Following feedback from GPs and children and adolescents on the content and understanding, the tool was piloted and implemented in general practice. A cohort of approximately 300 children and adolescents (age 8-19 years old) is being recruited from general practices (recruitment period, July 2019 - June 2020). Clinically meaningful risk groups (e.g. low/medium/high) for the recurrence/persistence of knee pain (at 3 and 6 months) will be identified. Discussion: If successful, this prognostic tool will allow GPs to gain insights into the likely prognosis of adolescents with knee pain and subsequently provide the first building blocks towards stratified care, where treatments will be matched to the patients' prognostic profile. This has the potential to improve the recovery of children and adolescents from knee pain, to improve the allocation of resources in primary care, and to avoid the decline in physical activity and potential associated health and social consequences due to adolescent knee pain. Registration: Registered with ClinicalTrials.gov on 24 June 2019 (ID NCT03995771).


Study of the electronic structure of electron accepting cyano-films: TCNQversusTCNE.
In this article, we perform systematic research on the electronic structure of two closely related organic electron acceptor molecules (TCNQ and TCNE), which are of technological interest due to their outstanding electronic properties. These studies have been performed from the experimental point of view by the use electron spectroscopies (XPS and UPS) and supported theoretically by the use of ab-initio DFT calculations. The cross-check between both molecules allows us to identify the characteristic electronic features of each part of the molecules and their contribution to the final electronic structure. We can describe the nature of the band gap of these materials, and we relate this with the appearance of the shake-up features in the core level spectra. A band bending and energy gap reduction of the aforementioned electronic structure in contact with a metal surface are seen in the experimental results as well in the theoretical calculations. This behavior implies that the TCNQ thin film accepts electrons from the metal substrate becoming a Schottky n-junction.


Comparison of the care of children with Down's syndrome with the care of matched controls.
A prospective study of the care of 134-children with Down's syndrome and 134 age- and sex-matched control children during 1981 has shown that the former group had significantly greater contact with the general practitioner, mostly owing to respiratory problems which were treated significantly more often with antibiotics. Referrals to specialist care were more common in the Down's children but the interface between general practice and paediatric care was not great. The study emphasizes the need for general practitioners to plan the care of Down's children and normal children with respect both to acute illness and the monitoring of chronic childhood illness.


Alcohol and acetaldehyde metabolism in Caucasians, Chinese and Amerinds.
Ethanol (0.4 to 0.8 g/kg in 30 minutes) was given by mouth to 102 healthy young volunteers (37 Caucasian men, 21 Caucasian women, 20 Chinese men and 24 Ojibwa men). Venous blood concentrations of ethanol and acetaldehyde 60, 90, 120 and 150 minutes after the end of drinking were measured by gas chromatography. The calculated rates of ethanol metabolism in the Caucasian men and women did not differ, but the overall group means for subgroups of Caucasians (103.6 mg/kg-h), Chinese (136.6 mg/kg-h) and Ojibwa (182.7 mg/kg-h) with decreasing postabsorption values differed significantly from each other. Mean acetaldehyde values paralleled the rates of ethanol metabolism: Ojibwa, 14.6 mug/ml; Chinese, 10.0 mug/ml; and Caucasians, 9.4 mug/ml. The high rate of ethanol metabolism in Amerind subjects differs from previous findings. Habitual level of alcohol consumption, proportion of body fat and genetic factors appear to account for most of the group differences.


The roles of sexual and asexual reproduction in the origin and dissemination of strains causing fungal infectious disease outbreaks.
Sexual reproduction commonly refers to the reproductive process in which genomes from two sources are combined into a single cell through mating and then the zygote genomes are partitioned to progeny cells through meiosis. Reproduction in the absence of mating and meiosis is referred to as asexual or clonal reproduction. One major advantage of sexual reproduction is that it generates genetic variation among progeny which may allow for faster adaptation of the population to novel and/or stressful environments. However, adaptation to stressful or new environments can still occur through mutation, in the absence of sex. In this review, we analyzed the relative contributions of sexual and asexual reproduction in the origin and spread of strains causing fungal infectious diseases outbreaks. The necessity of sex and the ability of asexual fungi to initiate outbreaks are discussed. We propose a framework that relates the modes of reproduction to the origin and propagation of fungal disease outbreaks. Our analyses suggest that both sexual and asexual reproduction can play critical roles in the origin of outbreak strains and that the rapid spread of outbreak strains is often accomplished through asexual expansion.


Development of nanosecond spike pulse power supply for electrochemical micromachining.
The use of pulse voltage can greatly improve the precision of electrochemical microfabrication, and the narrower the pulse width of the applied pulse voltage signal, the higher the machining precision. However, the commonly used chopper circuit topology of pulse power supplies is limited by the maximum switching frequency of the field-effect transistor. To address this problem, this paper proposes a nanosecond pulse electrochemical micromachining power supply based on a differential circuit. The power supply uses the STM32F103C8T6 microcontroller as the control core to output high-performance rectangular waves through a DDS device. After differential, rectification, filtering, and power amplification processing, stable, frequency, amplitude, and pulse width adjustable spike pulse voltage signals are obtained. By establishing a system mathematical model and optimizing the time constant of the differential circuit, theoretically, the sub-nanosecond pulse width can be obtained. Prototype performance tests show that the power supply has a maximum frequency of 20 MHz, a minimum pulse width of 1.8 ns, and a maximum peak voltage of 10 V. By using this power supply for microhole electrochemical machining experiments, nanometer-level machining precision has been achieved.


Oxidative capacity distribution in skeletal muscle fibers of the rat.
To study the distribution of oxidative capacity in muscle fibers, mitochondrial volume density and the oxidative capacity of isolated mitochondria were evaluated. Mitochondria were isolated from the subsarcolemmal and interfibrillar areas of the soleus (a muscle largely made up of slow oxidative fibers) and the gastrocnemius medial head (a muscle largely made up of fast glycolytic fibers) of the rat, and their oxidative capacities were evaluated using NADH- and FADH-generating substrates. In the soleus muscle, the subsarcolemmal mitochondria showed a lower oxidative capacity than interfibrillar mitochondria when NADH-generating substrates were used. This difference was not observed when FADH-generating substrates were used. In the gastrocnemius, there were no differences in the oxidative capacity of the subsarcolemmal and the interfibrillar mitochondria. Additionally, citrate synthase activity was found to be lower in mitochondria isolated from the subsarcolemmal area of the soleus than in the other mitochondrial preparations. These findings indicate that the difference in oxidative capacity of the isolated mitochondria is not related to differences in the inner mitochondrial membranes. Mitochondrial volume density was evaluated using electron micrographs of the subsarcolemmal and interfibrillar areas of slow oxidative fibers from the soleus and fast glycolytic fibers from the gastrocnemius. In the slow oxidative fibers, mitochondrial volume density in the subsarcolemmal area was four times higher than in the interfibrillar area. In the fast glycolytic fibers, mitochondrial volume densities in the subsarcolemmal and interfibrillar areas did not differ from that of the interfibrillar area of the slow oxidative fibers. The oxidative capacity of the tissue, calculated by multiplying the mitochondrial oxidative capacities by the mitochondrial volume densities, was 2-4 times higher in the subsarcolemmal areas of the soleus fibers than in the other areas studied. This was true in spite of the fact that the oxidative capacity of the subsarcolemmal mitochondria of the slow oxidative fibers was lower than those of the other mitochondrial populations studied. These results indicate that the difference in oxidative capacity between slow oxidative fibers and fast glycolytic fibers is the result of the much greater mitochondrial volume density in the subsarcolemmal area of the slow oxidative fibers.


[Ultrasonographic assessment of infantile hip by the Graf method].
Ultrasonography of the hip performed by the Graf method is widely accepted as an important diagnostic and treatment tool for hip dysplasia. This study aims to examine the reliability and reproducibility of the interpretations of ultrasound scans by general orthopedists. Thirteen examiners evaluated five hip ultrasounds of the same normal child. The examiners measured alpha and beta angles according to Graf. Statistical analysis was performed by variance analysis (ANOVA) and agreement was calculated and expressed as the intraclass correlation coefficient (ICC). The alpha angle mean was 37.32 degrees (11.12) and the beta angle mean was 45.05 degrees (10.49). Error of the intra-observer was 17.65 for the alpha angle and 17.63 for the beta. Error of the inter-observer was 7.68 for the alpha angle and 17.21 for the beta. ICC intra-observer was 0.18 for the alpha angle and 0.29 for the beta. ICC inter-observer was -0.02 for the alpha angle and 0.3 for the beta. Ultrasound of the infantile hip using the Graf method is an important tool for the assessment of developmental dysplasia of the hip, however, it must be emphasized that preciseness of interpretation requires training and frequent use.


Differential interaction of the E3 ligase parkin with the proteasomal subunit S5a and the endocytic protein Eps15.
Parkin is a multidomain E3 ligase associated with autosomal recessive Parkinson disease. The N-terminal ubiquitin-like domain (Ubld) of parkin functions with the S5a proteasomal subunit, positioning substrate proteins for degradation. In addition the parkin Ubld recruits the endocytotic protein Eps15, allowing the E3 ligase to ubiquinate Eps15 distal from its parkin-interacting site. The recognition sequences in the S5a subunit and Eps15 for the parkin Ubld are ubiquitin-interacting motifs (UIM). Each protein has two UIM sequences separated by a 50-residue spacer in S5a, but only approximately 5 residues in Eps15. In this work we used NMR spectroscopy to determine how the parkin Ubld recognizes the proteasomal subunit S5a compared with Eps15, a substrate for ubiquitination. We show that Eps15 contains two flexible alpha-helices each encompassing a UIM sequence. The alpha-helix surrounding UIM II is longer than that for UIM I, a situation that is reversed from S5a. Furthermore, we show the parkin Ubld preferentially binds to UIM I in the S5a subunit. This interaction is strongly diminished in a K48A substitution, found near the center of the S5a interacting surface on the parkin Ubld. In contrast to S5a, parkin recruits Eps15 using both its UIM sequences resulting in a larger interaction surface that includes residues from beta1 and beta2, not typically known to interact with UIM sequences. These results show that the parkin Ubld uses differential surfaces to recruit UIM regions from the S5a proteasomal subunit compared with Eps15 involved in cell signaling.


Age of natural menopause and atrial fibrillation: the Framingham Heart Study.
Early menopausal age is associated with risk of cardiovascular events including myocardial infraction, stroke, and increased mortality. Relations between menopausal age and atrial fibrillation (AF) have not been investigated. We examined the association between menopausal age and AF. Framingham Heart Study women ≥ 60 years old without prevalent AF and natural menopause were followed up for 10 years or until incident AF. Menopausal age was modeled as a continuous variable and by categories (<45, 45-53, and >53 years). We used Cox proportional hazards regression to determine associations between menopausal age and AF risk. In 1,809 Framingham women (2,662 person-examinations, mean baseline age 71.4 ± 7.6 years, menopausal age 49.8 ± 3.6 years), there were 273 unique participants with incident AF. We did not identify a significant association between the SD of menopausal age (3.6 years) and AF (hazard ratio [HR] per SD 0.94, 95% CI 0.83-1.06; P = .29). In a multivariable model with established risk factors for AF, menopausal age was not associated with incident AF (HR per SD 0.97, 95% CI 0.86-1.09; P = .60). Examining categorical menopausal age, earlier menopausal age (<45 years) was not significantly associated with increased AF risk compared with older menopausal age >53 years (HR 1.20, 95% CI 0.74-1.94; P = .52) or menopausal age 45 to 53 years (HR 1.38, 95% CI 0.93-2.04; P = .11). In our moderate-sized, community-based sample, we did not identify menopausal age as significantly increasing AF risk. However, future larger studies will need to examine whether there is a small effect of menopausal age on AF risk.


Individual dairy cow management: achievements, obstacles and prospects.
This review deals with the prospects and achievements of individual dairy cow management (IDCM) and the obstacles and difficulties encountered in attempts to successfully apply IDCM into routine dairy management. All aspects of dairy farm management, health, reproduction, nutrition and welfare are discussed in relation to IDCM. In addition, new IDCM R&D goals in these management fields are suggested, with practical steps to achieve them. The development of management technologies is spurred by the availability of off-the-shelf sensors and expanded recording capacity, data storage, and computing capabilities, as well as by demands for sustainable dairy production and improved animal wellbeing at a time of increasing herd size and milk production per cow. Management technologies are sought that would enable the full expression of genetic and physiological potential of each cow in the herd, to achieve the dairy operation's economic goals whilst optimizing the animal's wellbeing. Results and conclusions from the literature, as well as practical experience supported by published and unpublished data are analyzed and discussed. The object of these efforts is to identify knowledge and management routine gaps in the practical dairy operation, in order to point out directions and improvements for successful implementation of IDCM in the dairy cows' health, reproduction, nutrition and wellbeing.


Who needs a blood culture? A prospectively derived and validated prediction rule.
The study objective was to derive and validate a clinical decision rule for obtaining blood cultures in Emergency Department (ED) patients with suspected infection. This was a prospective, observational cohort study of consecutive adult ED patients with blood cultures obtained. The study ran from February 1, 2000 through February 1, 2001. Patients were randomly assigned to derivation (2/3) or validation (1/3) sets. The outcome was "true bacteremia." Features of the history, co-morbid illness, physical examination, and laboratory testing were used to create a clinical decision rule. Among 3901 patients, 3730 (96%) were enrolled with 305 (8.2%) episodes of true bacteremia. A decision rule was created with "major criteria" defined as: temperature > 39.5 degrees C (103.0 degrees F), indwelling vascular catheter, or clinical suspicion of endocarditis. "Minor criteria" were: temperature 38.3-39.4 degrees C (101-102.9 degrees F), age > 65 years, chills, vomiting, hypotension (systolic blood pressure < 90 mm Hg), neutrophil% > 80, white blood cell count > 18 k, bands > 5%, platelets < 150 k, and creatinine > 2.0. A blood culture is indicated by the rule if at least one major criterion or two minor criteria are present. Otherwise, patients are classified as "low risk" and cultures may be omitted. Only 4 (0.6%) low-risk patients in the derivation set and 3 (0.9%) low-risk patients in the validation set had positive cultures. The sensitivity was 98% (95% confidence interval [CI] 96-100%) (derivation) and 97% (95% CI 94-100%) (validation). We developed and validated a promising clinical decision rule for predicting bacteremia in patients with suspected infection.


Multifaceted interactions of bacterial toxins with the gastrointestinal mucosa.
The digestive tract is one of the ecosystems that harbors the largest number and greatest variety of bacteria. Among them, certain bacteria have developed various strategies, including the synthesis of virulence factors such as toxins, to interact with the intestinal mucosa, and are responsible for various pathologies. A large variety of bacterial toxins of different sizes, structures and modes of action are able to interact with the gastrointestinal mucosa. Some toxins, termed enterotoxins, directly stimulate fluid secretion in enterocytes or cause their death, whereas other toxins pass through the intestinal barrier and disseminate by the general circulation to remote organs or tissues, where they are active. After recognition of a membrane receptor on target cells, toxins can act at the cell membrane by transducing a signal across the membrane in a hormone-like manner, by pore formation or by damaging membrane compounds. Other toxins can enter the cells and modify an intracellular target leading to a disregulation of certain physiological processes or disorganization of some structural architectures and cell death. Toxins are fascinating molecules, which mimic or interfere with eukaryotic physiological processes. Thereby, they have permitted the identification and characterization of new natural hormones or regulatory pathways. Besides use as protective antigens in vaccines, toxins offer multiple possibilities in pharmacology, such as immune modulation or specific delivery of a protein of interest into target cells.


Evaluation of the Prevalence of Drug Abuse and Smoking in Parents of Children with Attention Deficit Hyperactivity Disorder.
Attention deficit hyperactivity disorder (ADHD) affects 5% of children. In addition to pharmacotherapy, non-drug treatments such as appropriate parenting are also very important in the treatment of these children. Diagnosis and treatment of parents with psychiatric disorders and substance abuse and evaluation of the frequency of these disorders in parents is critical. In this case-control study, 200 parents were studied. The target population included parents of 7 to 12 year-old children who referred to child and adolescent psychiatric clinics. The control group included parents of children who referred to child non-psychiatric clinics. The parents were evaluated via a demographic information form, and structured interviews based on the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV) for nicotine and drug addiction. Then, the Hamilton Anxiety Rating Scale (HAM-A) and Hamilton Depression Rating Scale (HDRS) was used to assess their anxiety and depression. Among the studied parents, the comparison of drug abuse, smoking, and stress showed significant differences between the two groups. In terms of depression and ADHD, the difference between the case and control groups was not statistically significant. The higher prevalence of these disorders in parents of children with ADHD may indicate the possible role of this disorder in the etiology.


Women's experience of the decision-making process for home-based postnatal midwifery care when discharged early from hospital: A Swedish interview study.
Women and their families are often excluded from reproductive decision-making processes in postnatal care, and do not know which choices they have. Shared decision-making is a critical but challenging component of maternity care quality. The aim was to explore women's experience of the decision-making process about early return from hospital with home-based postnatal midwifery care. This is a descriptive qualitative study. In total, 24 women participated in a semi-structured telephone interview, averaging 58 minutes. Data were analyzed using thematic analysis according to Braun and Clarke. The main theme explored was 'The supremacy of giving new mothers autonomy to decide on the postnatal care model they would prefer'. Important aspects of the women's decision-making process were the time-point for receiving information about the home-based midwifery model of care, to receive sufficient time for consideration about the model, to have a rationale for choosing home-based care, and to comprehend the concept. Women must be given sufficient time for consideration and necessary information about postnatal care models, which is essential for making an informed decision. Parents' readiness for discharge must be identified by midwives who need to facilitate shared decision-making by introducing early postnatal care model choices, describe these options, and support women to explore their preferences. Midwives must ensure parents' participation in decision-making for the time of discharge from hospital.


HRQOLISP-26: A Concise, Multiculturally Valid, Multidimensional, Flexible, and Reliable Stroke-Specific Measure.
Background. A multidimensional, brief, and flexible stroke-specific health-related quality of life (HRQOL) measure is still needed. The aim was to develop a shortened version of the HRQOLISP-102, a multiculturally generated measure with excellent psychometric properties. Methods. Participants included 100 (Ibadan, Nigeria) and 103 (Berlin, Germany) stroke patients compared to 100 (Ibadan) and 50 (Berlin) apparently healthy adults. Using standard protocol, the 26-item version was generated, consisting of therapeutically relevant physical, psychological, cognitive, and ecosocial domains. Criterion validity of the HRQOLISP-26 was determined using Bland-Altman statistics. "Known groups" validity was assessed using NIHSS, stroke levity score, and modified Rankin scale. Results. HRQOLISP-26 was easily interpretable and precise with no significant floor/ceiling effect. It can be completed within 7 minutes. It showed good content, construct, "known groups," and criterion validity. It demonstrated good internal consistency (α = 0.81, 0.89) and test-retest reliability. Conclusions. HRQOLISP-26 is novel, brief, multiculturally-valid, and flexible for routine assessment of HRQOL in stroke patients.


Environmental Noise-Induced Effects on Stress Hormones, Oxidative Stress, and Vascular Dysfunction: Key Factors in the Relationship between Cerebrocardiovascular and Psychological Disorders.
The role of noise as an environmental pollutant and its adverse effects on health are being increasingly recognized. Beyond its direct effects on the auditory system (e.g., hearing loss and tinnitus induced by exposure to high levels of noise), chronic low-level noise exposure causes mental stress associated with known cardiovascular complications. According to recent estimates of the World Health Organization, exposure to traffic noise is responsible for a loss of more than 1.5 million healthy life years per year in Western Europe alone, a major part being related to annoyance, cognitive impairment, and sleep disturbance. Underlying mechanisms of noise-induced mental stress are centered on increased stress hormone levels, blood pressure, and heart rate, which in turn favor the development of cerebrocardiovascular disease such as stroke, arterial hypertension, ischemic heart disease, and myocardial infarction. Furthermore, traffic noise exposure is also associated with mental health symptoms and psychological disorders such as depression and anxiety, which further increase maladaptive coping mechanisms (e.g., alcohol and tobacco use). From a molecular point of view, experimental studies suggest that traffic noise exposure can increase stress hormone levels, thereby triggering inflammatory and oxidative stress pathways by activation of the nicotinamide adenine dinucleotide phosphate oxidase, uncoupling of endothelial/neuronal nitric oxide synthase inducing endothelial and neuronal dysfunction. This review elucidates the mechanisms underlying the relationship between noise exposure and cerebrocardiovascular and psychological disorders, focusing on mental stress signaling pathways including activation of the autonomous nervous system and endocrine signaling and its association with inflammation, oxidative stress, and vascular dysfunction.


A catalytic mechanism for cysteine N-terminal nucleophile hydrolases, as revealed by free energy simulations.
The N-terminal nucleophile (Ntn) hydrolases are a superfamily of enzymes specialized in the hydrolytic cleavage of amide bonds. Even though several members of this family are emerging as innovative drug targets for cancer, inflammation, and pain, the processes through which they catalyze amide hydrolysis remains poorly understood. In particular, the catalytic reactions of cysteine Ntn-hydrolases have never been investigated from a mechanistic point of view. In the present study, we used free energy simulations in the quantum mechanics/molecular mechanics framework to determine the reaction mechanism of amide hydrolysis catalyzed by the prototypical cysteine Ntn-hydrolase, conjugated bile acid hydrolase (CBAH). The computational analyses, which were confirmed in water and using different CBAH mutants, revealed the existence of a chair-like transition state, which might be one of the specific features of the catalytic cycle of Ntn-hydrolases. Our results offer new insights on Ntn-mediated hydrolysis and suggest possible strategies for the creation of therapeutically useful inhibitors.


Current status of immunological approaches for the treatment of prostate cancer.
The recent Food and Drug Administration approval of sipuleucel-T for metastatic castration-resistant prostate cancer and of the anticytotoxic T-lymphocyte antigen 4 antibody (Ipilimumab) for metastatic melanoma has led to a renewed interest in immunotherapy for prostate and other cancers. Ipilimumab has entered phase III testing for prostate cancer, as has a viral-based anti-prostate-specific antigen vaccine (ProstVac-VF). Complementing these phase III studies are a number of innovative phase II studies, aimed at bringing immunotherapy forward in the setting of less advanced disease, as well as a number of interesting trials combining immunotherapy with conventional therapy for prostate cancer. Although a number of immunotherapy trials have been initiated, few mature results are available at the current time. These data are likely to mature in the setting of an increasingly complex treatment paradigm in which multiple hormonal and novel agents are available. Immunotherapy for prostate cancer represents an attractive treatment approach, with the currently available agent sipuleucel-T providing a significant survival benefit without appreciable toxicity. Novel approaches to improve the efficacy of this and other immune-active agents are currently under evaluation.


A quantitative method for classification of EEG in the fetal baboon.
Electroencephalographic (EEG) activity is used as a primary indicator of sleep states in adults and infants of many species and in the ovine fetus. We recently reported that the baboon fetus exhibits visually discernable patterns of EEG activity. One pattern of activity, characterized by the intermittent presence of repetitive bursts of high-voltage EEG, is indistinguishable from trace alternant (TA). TA is a distinctive pattern of EEG activity found only during early stages of development in primates. TA is the predominant pattern of EEG activity during quiet sleep in human infants < 2 mo of age. The focus of this study was to derive quantitative parameters that would discriminate TA from other activity and then to develop a method for automated categorization of EEG patterns. Results demonstrate that several parameters derived from frequency-domain analyses are related to visually coded EEG states. Among these parameters, high-frequency power (12-24 Hz) and spectral-edge frequency are good discriminators of EEG patterns. This paper describes a new parameter, EEG ratio, computed as spectral power in the rectified EEG within a band that corresponds to the frequency of bursts of activity during TA (0.03-0.20 Hz) divided by power in the 12- to 24-Hz band. This new composite parameter of EEG activity provides a markedly better correlate of visually coded EEG than any of the individual parameters tested. Using cluster analysis, we devised a method for objective minute-by-minute dichotomization of EEG ratio. The method produces results that agree with visual coding of EEG activity 87.1% of the time.(ABSTRACT TRUNCATED AT 250 WORDS)


Sequence diversity of V1 and V2 domains of gp120 from human immunodeficiency virus type 1: lack of correlation with viral phenotype.
We analyzed by PCR and direct sequencing 57 viral sequences from 47 individuals infected with human immunodeficiency virus type 1, focussing on the V1 and V2 regions of gp120. There was extensive length polymorphism in the V1 region, which rendered sequence alignment difficult. The V2 hypervariable locus also displayed considerable length variations, whereas flanking regions were relatively conserved. Two-thirds of the amino acid residues in these flanking regions were highly conserved (> 80%), presumably reflecting their critical contribution to V2 structure or function. We also characterized the syncytium-inducing properties of the isolates from which we derived sequence information. There was no correlation between V1 or V2 sequences and the viral phenotype, contrary to a previous report (M. Groenink, R. A. M. Fouchier, S. Broersen, C. H. Baker, M. Koot, A. B. van't Wout, H. G. Huisman, F. Miedema, M. Tersmette, and H. Schuitemaker, Science 260:1513-1516, 1993). The sequence heterogeneity described in this study provides information to suggest that it would be most difficult to exploit the V1 and V2 domains for vaccine development.


[Changes in maternal stress with the age of her developmentally disabled child--comparison between mentally retarded and autistic children using cross-sectional data].
In order to explore a transitional pattern of mother's psycho-social stress accompanying the age of developmentally-disabled child, we analized the stress patterns obtained from 1 036 mothers of mental retarded and autistic children by means of 10 stress scales consisted of four or five items for each scale. Mothers of 3 to 15 yrs old children, classified by cross-sectional method, were divided by their child's age into seven groups, and multiple-range tests were applied to the mean stress scores of mothers in the seven groups for each of two disability. In the results, we found three kinds of transitional patterns of stress. They were named as follows: No change type, Decreased type, and increased-at-first, but later-decreased type. These transitional patterns are common to two disabilities in the same scale, and furthermore it is worthy of notice that the stage of changing point of increase or decrease of the amount of stress coincides with the periods of child's entrance to primary school and/or junior high school at almost all scales.


Glucose: archetypal biomarker in diabetes diagnosis, clinical management and research.
The clinical utility of diabetes biomarkers can be considered in terms of diagnosis, management and prediction of long-term vascular complications. Glucose satisfies all of these requirements. Thresholds of hyperglycemia diagnostic of diabetes reflect inflections that confer a risk of developing long-term microvascular complications. Degrees of hyperglycemia (impaired fasting glucose, impaired glucose tolerance) that lie below the diagnostic threshold for diabetes identify individuals at risk of progression to diabetes and/or development of atherothrombotic cardiovascular disease. Self-measured glucose levels usefully complement hemoglobin A1c levels to guide daily management decisions. Continuous glucose monitoring provides detailed real-time data that is of value in clinical decision making, assessing response to new diabetes drugs and the development of closed-loop artificial pancreas technology.


Effects of perinatal oxycodone exposure on the cardiovascular response to acute stress in male rats at weaning and in young adulthood.
Oxycodone (OXY) is one of the most commonly abused opiates during pregnancy. Perinatal opiate exposure (POE) is associated with neurobehavioral and hormone changes. Little is known about the effects of perinatal OXY on the cardiovascular (CV) responses to stress. to determine the effects of POE on: (1) CV responses to acute stress and ability to discriminate using a classical conditioning paradigm; (2) changes in CV response to the paradigm and retention of the ability to discriminate from postnatal day (PD) 40 to young adulthood. Pregnant rats were given i.v. OXY or vehicle (CON) daily. OXY and CON males were fitted with BP telemetry units. Offspring were classically conditioned by following a pulsed tone (CS+) with tail shock. A steady tone (CS-) was not followed by shock. BP and HR were recorded during resting periods and conditioning. Changes in BP, HR from composite analysis were compared. The paradigm was repeated on PD 75. At PD 40, OXY rats had a lower baseline mean BP (OXY: 114.8 ± 1.0 vs. CON: 118.3 ± 1.0 mm Hg; mean ± SEM) but larger amplitude of the conditional BP increase during the stress response (OXY: +3.9 ± 0.4 vs. CON: +1.7 ± 0.4 mm Hg). Both OXY and CON rats were able to discriminate between CS+ and CS-. At PD 75, the effects of OXY on the increased amplitude of the conditional BP had dissipated (CON: +3.4 ± 2.3 vs. OXY: +4.5 ± 1.4 mm Hg). BP responses to the stress and non-stress stimuli did not differ in the OXY group, suggesting that OXY may have decreased the ability of the offspring to discriminate (OXY: CS+: 147.1 ± 1.6, CS-: 145.9 ± 1.6 mm Hg vs. CON: CS+: 155.4 ± 2.7, CS-: 147.8 ± 2.7 mm Hg). POE is associated with subtle alterations in stress CV responses in weanling rats which dissipate when the conditioning is repeated at an early adult age. Although POE effect on the ability to discriminate at weanling age could not be detected, POE may impair retention of this ability in adulthood.


Mitogen-activated protein kinase phosphatase-1 in human breast cancer independently predicts prognosis and is repressed by doxorubicin.
Mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) dephosphorylates mitogen-activated protein kinase [extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38], mediates breast cancer chemoresistance, and is repressible by doxorubicin in breast cancer cells. We aimed to characterize doxorubicin effects on MKP-1 and phospho-MAPKs in human breast cancers and to further study the clinical relevance of MKP-1 expression in this disease. Doxorubicin effects on MKP-1, phospho-ERK1/2 (p-ERK1/2), phospho-JNK (p-JNK), and phospho-p38 were assayed in a panel of human breast cancer cells by Western blot and in human breast cancer were assayed ex vivo by immunohistochemistry (n = 50). MKP-1 expression was also assayed in a range of normal to malignant breast lesions (n = 30) and in a series of patients (n = 96) with breast cancer and clinical follow-up. MKP-1 was expressed at low levels in normal breast and in usual ductal hyperplasia and at high levels in in situ carcinoma. MKP-1 was overexpressed in approximately 50% of infiltrating breast carcinomas. Similar to what was observed in breast cancer cell lines, ex vivo exposure of breast tumors to doxorubicin down-regulated MKP-1, and up-regulated p-ERK1/2 and p-JNK, in the majority of cases. However, in a proportion of tumors overexpressing MKP-1, doxorubicin did not significantly affect MKP-1 or phospho-MAPKs. With regard to patient outcome, MKP-1 overexpression was an adverse prognostic factor for relapse both by univariate (P < 0.001) and multivariate analysis (P = 0.002). MKP-1 is overexpressed during the malignant transformation of the breast and independently predicts poor prognosis. Furthermore, MKP-1 is repressed by doxorubicin in many human breast cancers.


A method for familiarizing nurses with the hospital formulary.
A slide-tape audiovisual program was developed by a university-based drug information center for the purpose of familiarizing the nursing staff with our institution's formulary. The formulary contains useful information on medications stocked by our pharmacy, including costs, dosage forms, policies and procedures governing the pharmacy department, and many useful tables and figures. It was felt that the nursing staff was not aware of these components and thus, was not optimally utilizing the formulary. Nurses taking a test after the slide-tape presentation scored higher on the average than nurses from similar nursing stations who had not seen the presentation. These results, as well as the positive subjective responses from the individual nurses regarding the presentation, indicate that the slide-tape format is an effective means of acquainting the nursing staff with the hospital formulary. It is anticipated that this new awareness of the formulary will result in its more efficient use and further promote the pharmacy department as an information provider.


Motivators for participation in a whole-genome sequencing study: implications for translational genomics research.
The promise of personalized medicine depends on the ability to integrate genetic sequencing information into disease risk assessment for individuals. As genomic sequencing technology enters the realm of clinical care, its scale necessitates answers to key social and behavioral research questions about the complexities of understanding, communicating, and ultimately using sequence information to improve health. Our study captured the motivations and expectations of research participants who consented to participate in a research protocol, ClinSeq, which offers to return a subset of the data generated through high-throughput sequencing. We present findings from an exploratory study of 322 participants, most of whom identified themselves as white, non-Hispanic, and coming from higher socio-economic groups. Participants aged 45-65 years answered open-ended questions about the reasons they consented to ClinSeq and about what they anticipated would come of genomic sequencing. Two main reasons for participating were as follows: a conviction to altruism in promoting research, and a desire to learn more about genetic factors that contribute to one's own health risk. Overall, participants expected genomic research to help improve understanding of disease causes and treatments. Our findings offer a first glimpse into the motivations and expectations of individuals seeking their own genomic information, and provide initial insights into the value these early adopters of technology place on information generated by high-throughput sequencing studies.


[Equilibrium angioscintigraphic evaluation of sequential changes in right ventricle function in patients with lower myocardial infarct and ischemic co-involvement of the right ventricle: long-term study].
The purpose of this study was to define the sequential changes in global and regional right ventricular function, using equilibrium gated radionuclide angiography, following transmural inferior myocardial infarction (IMI) and associated ischemic right ventricular involvement. 24 patients with IMI underwent radionuclide angiography within 72 hours of onset of chest pain; subsequent studies were done at 13 +/- 5 days and at 6 +/- 2 months. Scintigraphic evidence of ischemic right ventricular involvement was defined by depression of right ventricular ejection fraction (less than 40%) and regional wall motion abnormalities (hypo-a-dyskinesia of right ventricular free wall). Significant improvement of global right ventricular ejection fraction was observed in 15 patients at the second study, and in 21 patients at the third study. Right ventricular regional wall motion showed similar improvement from the initial to the final studies. Significant changes in right ventricular function occurred without concurrent changes in global and regional left ventricular ejection fraction. We concluded that patients with IMI and ischemic right ventricular involvement show frequently improvement of global and regional right ventricular function over time. This changes tend to occur early and without significant modifications in left ventricular function. The good prognosis observed in our patients, despite the high incidence of in hospital complications, might be related to the improvement of right ventricular function.


Validation of cognitive and neuromuscular latencies using an iPad-based tool to quantify simple reaction times in asymptomatic subjects.
Simple reaction times (SRTs), measured in milliseconds (msec), are equal to the sum of subject-dependent latencies that occur during cognitive processing and neuromuscular responses to a preprogrammed stimulus presented to a subject. SRTs have the advantage of being a relatively pure, neurologically driven motor/sensory task that provides a clinician with a generalized assessment of functional deficits. SRTs are easily obtained, and studies have reported that the average number of finger-taps during a 10-s interval can be utilized to distinguish between patients with mild traumatic brain injury (TBI) and healthy controls. A stimulus/response SRT protocol, utilizing an iPad-based tool, was utilized to see if differentiation and quantification of individual components of cognitive latency (CL) and neuromuscular latency (NL) from a subject's total SRT could be accomplished. The study hypothesis was that cognitive latencies, related to a specific cognitive challenge, would remain constant even as latencies due to neuromuscular challenges changed. Two categories of SRT tests were utilized. The first was a simple finger tapping test (FTT), without any cognitive involvement, that was designed to quantify pure NL. The second test was a choice test (CT) that was characterized by the addition of a cognitive task to the FTT. The objective of the FTT was for the subject to simply tap a single target positioned on the iPad screen as fast as possible over an interval of 10 s. Measurement of the SRT began when a target was displayed and ended when the target was tapped. Two levels of challenge for the FTT and CT tests were presented to the subjects: a small random displacement (SD) of the target of up to 6 mm, and a large random displacement (LD) of the target of up to 24 mm. It was expected that the magnitude of the SRT (FTT) would be directly related to the magnitude of the displacement of the target due to kinematic response characteristics of the hand and arm. To validate the study hypothesis, CL for both small and large displacements was calculated by subtracting the respective NL from the total SRT. Utilizing a repeated measures t-test analysis utilizing SPSS, a significant difference between CL (SD) and CL (LD) at p=0.696 was not observed. Differentiation and quantification of individual components of CL and NL from a subject's total SRT was accomplished. An Apple iPad Pro was selected as a platform for our study because: (1) it is readily available, affordable, and programmable; (2) it meets a requirement for portability; and (3) it allows the modification/addition of test parameters to meet future needs. The ability to quantify the extent of cognitive and neuromuscular dysfunction in the TBI patient is an essential component of developing an effective treatment plan.


Amyloid positron emission tomography candidates may focus more on benefits than risks of results disclosure.
Given mounting calls to disclose biomarker test results to research participants, we explored factors underlying decisions by patients with mild cognitive impairment to receive amyloid imaging results. Prospective, qualitative interviews were conducted with 59 participants (30 = mild cognitive impairment patients, 29 = care partners) from the scan arm of a randomized controlled trial on the effects of amyloid PET results disclosure in an Alzheimer Disease Research Center setting. Sixty-three percent of the participants were female, with an average age of 72.9 years, and most had greater than a high school level of education (80%). Primary motivations included: (1) better understanding one's mild cognitive impairment etiology and prognosis to plan ahead, and (2) learning one's brain amyloid status for knowledge's sake, regardless of whether the information is actionable. Most participants demonstrated an adequate understanding of the scan's limitations, yet instances of characterizing amyloid PET as a definitive test for Alzheimer's disease occurred. Mention of potential drawbacks, such as negative psychological outcomes, was minimal, even among care partners. Findings demonstrate a risk of disproportionate focus on possible benefits of testing among amyloid scan candidates and suggest a need to clearly emphasize the limitations of amyloid PET when counseling cognitively impaired patients and their families before testing. Future research should examine whether minimizing drawbacks at the pre-imaging stage has adverse consequences on results disclosure.


Autologous bone marrow transplantation in high-risk remission T-lineage acute lymphoblastic leukemia using immunotoxins plus 4-hydroperoxycyclophosphamide for marrow purging.
Fourteen patients with high-risk T-lineage acute lymphoblastic leukemia (ALL) in complete remission underwent autologous bone marrow transplantation (BMT) in an attempt to eradicate their residual disease burden. A combined immunochemotherapy protocol using a cocktail of two immunotoxins directed against CD5/Tp67 and CD7/Tp41 T-lineage differentiation antigens in combination with the in vitro active cyclophosphamide congener 4-hydroperoxy-cyclophosphamide (4-HC) was used to purge autografts. Despite high dose pretransplant radiochemotherapy and effective purging of autografts, 9 of 14 patients relapsed at a median of 2.5 months (range, 1.2 to 16.8 months) post BMT. Two patients remain alive and disease free at 26 and 28 months post BMT. We used a novel quantitative minimal residual disease (MRD) detection assay, which combines fluorescence activated multiparameter flow cytometry and cell sorting with leukemic progenitor cell (LPC) assays, to analyze remission bone marrow (BM) samples from T-lineage ALL patients for the presence of residual LPCs. Notably, high numbers of residual LPC detected in remission BM before BMT constituted a poor prognostic indicator, providing the first evidence for the biologic significance and clinical value of in vitro T-lineage ALL LPC assays. The median value for the residual leukemia burden before BMT, was approximately 8.6 x 10(3) LPC/10(8) mononuclear cells (MNC) (approximately 0.0086% LPC). Patients with a residual leukemia burden less than this median value appeared to have a better outlook for remaining free of relapse after autologous BMT than patients with a greater leukemia burden (53 +/- 25% v 14 +/- 13%, P = .006, Mantel-Cox). By comparison, the log kill efficacy of purging, the remaining numbers of LPC in purged autografts, or the estimated numbers of reinfused LPC, did not correlate with the probability of disease-free survival (DFS). These results indicate that the primary reason for the recurrence of leukemia was inefficient pretransplant radiochemotherapy rather than inefficient purging of autografts.


Broad-spectrum XX and XY gonadal dysgenesis in patients with a homozygous L193S variant in PPP2R3C.
Homozygous and heterozygous variants in PPP2R3C are associated with syndromic 46,XY complete gonadal dysgenesis (Myo-Ectodermo-Gonadal Dysgenesis (MEGD) syndrome), and impaired spermatogenesis, respectively. This study expands the role of PPP2R3C in the aetiology of gonadal dysgenesis (GD). We sequenced the PPP2R3C gene in four new patients from three unrelated families. The clinical, laboratory, and molecular characteristics were investigated. We have also determined the requirement for Ppp2r3c in mice (C57BL6/N) using CRISPR/Cas9 genome editing. A homozygous c.578T>C (p.L193S) PPP2R3C variant was identified in one 46,XX girl with primary gonadal insufficiency, two girls with 46,XY complete GD, and one undervirilised boy with 46,XY partial GD. The patients with complete GD had low gonadal and adrenal androgens, low anti-Müllerian hormone, and high follicle-stimulating hormone and luteinizing hormone concentrations. All patients manifested characteristic features of MEGD syndrome. Heterozygous Ppp2r3c knockout mice appeared overtly normal and fertile. Inspection of homozygous embryos at 14.5, 9.5, and 8.5 days post coitum(dpc) revealed evidence of dead embryos. We conclude that loss of function of Ppp2r3c is not compatible with viability in mice and results in embryonic death from 7.5 dpc or earlier. Our data indicate the essential roles for PPP2R3C in mouse and human development. Germline homozygous variants in human PPP2R3C are associated with distinctive syndromic GD of varying severity in both 46,XY and 46,XX individuals.


Crystal structures of KPC-2 β-lactamase in complex with 3-nitrophenyl boronic acid and the penam sulfone PSR-3-226.
Class A carbapenemases are a major threat to the potency of carbapenem antibiotics. A widespread carbapenemase, KPC-2, is not easily inhibited by β-lactamase inhibitors (i.e., clavulanic acid, sulbactam, and tazobactam). To explore different mechanisms of inhibition of KPC-2, we determined the crystal structures of KPC-2 with two β-lactamase inhibitors that follow different inactivation pathways and kinetics. The first complex is that of a small boronic acid compound, 3-nitrophenyl boronic acid (3-NPBA), bound to KPC-2 with 1.62-Å resolution. 3-NPBA demonstrated a K(m) value of 1.0 ± 0.1 μM (mean ± standard error) for KPC-2 and blocks the active site by making a reversible covalent interaction with the catalytic S70 residue. The two boron hydroxyl atoms of 3-NPBA are positioned in the oxyanion hole and the deacylation water pocket, respectively. In addition, the aromatic ring of 3-NPBA provides an edge-to-face interaction with W105 in the active site. The structure of KPC-2 with the penam sulfone PSR-3-226 was determined at 1.26-Å resolution. PSR-3-226 displayed a K(m) value of 3.8 ± 0.4 μM for KPC-2, and the inactivation rate constant (k(inact)) was 0.034 ± 0.003 s(-1). When covalently bound to S70, PSR-3-226 forms a trans-enamine intermediate in the KPC-2 active site. The predominant active site interactions are generated via the carbonyl oxygen, which resides in the oxyanion hole, and the carboxyl moiety of PSR-3-226, which interacts with N132, N170, and E166. 3-NPBA and PSR-3-226 are the first β-lactamase inhibitors to be trapped as an acyl-enzyme complex with KPC-2. The structural and inhibitory insights gained here could aid in the design of potent KPC-2 inhibitors.


Functional human hepatocytes: isolation from small liver biopsy samples and primary cultivation with liver-specific functions.
Morphologically and functionally intact human hepatocytes were isolated from small liver biopsy samples weighing about 1-2 g by initial digestion with collagenase followed by repeated digestions with trypsin. The usual yield of hepatocytes was greater than 1 x 10(7) cells per g of liver sample and cell viability, as judged by dye exclusion test, was routinely over 90%. The isolated human hepatocytes showed intact morphology under scanning electron microscope. Formation of membrane protrusions upon phalloidin addition demonstrated that the actin in isolated hepatocytes was maintained with its structural integrity. The cultured human hepatocytes retained a variety of liver-specific functions which were similarly exhibited by rat hepatocytes isolated using the same procedure. The cultured human hepatocytes exhibited a specific cytochrome P-450 related enzyme activity, and active amino acid uptake that increased upon addition of hormones like glucagon and dexamethasone. Additionally, the cultured human hepatocytes synthesized DNA actively and, human serum albumin, and was found to be responsive to modulation by growth modulating hormones, cytokines and hepatotoxic agents. Based on the profile of activity described above, the presently established conditions for isolation and culturing of human hepatocytes demonstrate that functional liver cells can be obtained from small biopsied liver samples.