The effect of lactoferrin in aging: role and potential.
Aging is frequently accompanied by various types of physiological deterioration, which increases the risk of human pathologies. Global public health efforts to increase human lifespan have increasingly focused on lowering the risk of aging-related diseases, such as diabetes, neurodegenerative diseases, cardiovascular disease, and cancers. Dietary intervention is a promising approach to maintaining human health during aging. Lactoferrin (LF) is known for its physiologically pleiotropic properties. Anti-aging interventions of LF have proven to be safe and effective for various pharmacological activities, such as anti-oxidation, anti-cellular senescence, anti-inflammation, and anti-carcinogenic. Moreover, LF has a pivotal role in modulating the major signaling pathways that influence the longevity of organisms. Thus, LF is expected to be able to attenuate the process of aging and greatly ameliorate its effects.


Adenovirus-vectored African Swine Fever Virus Antigens Cocktail Is Not Protective against Virulent Arm07 Isolate in Eurasian Wild Boar.
: African swine fever (ASF) is a viral disease of domestic and wild suids for which there is currently no vaccine or treatment available. The recent spread of ASF virus (ASFV) through Europe and Asia is causing enormous economic and animal losses. Unfortunately, the measures taken so far are insufficient and an effective vaccine against ASFV needs to be urgently developed. We hypothesized that immunization with a cocktail of thirty-five rationally selected antigens would improve the protective efficacy of subunit vaccine prototypes given that the combination of fewer immunogenic antigens (between 2 and 22) has failed to elicit protective efficacy. To this end, immunogenicity and efficacy of thirty-five adenovirus-vectored ASFV antigens were evaluated in wild boar. The treated animals were divided into different groups to test the use of BioMize adjuvant and different inoculation strategies. Forty-eight days after priming, the nine treated and two control wild boar were challenged with the virulent ASFV Arm07 isolate. All animals showed clinical signs and pathological findings consistent with ASF. This lack of protection is in line with other studies with subunit vaccine prototypes, demonstrating that there is still much room for improvement to obtain an effective subunit ASFV vaccine.


Circadian variations of proopiocortin-related peptides in children with migraine.
Basal and throughout-the-day variations of B-lipotropin (BLPH), B-endorphin (BEP), ACTH and cortisol plasma levels were studied in seven prepubertal children who had been affected by common migraine for periods of 6-26 months and in six healthy volunteers. Despite normal cortisol concentrations, children with migraine show higher BLPH (15.1 +/- 2.7 fmol/ml, M +/- SE), ACTH (25 +/- 2.7) and BEP (9.1 +/- 1.1) levels than controls. In both groups of children, evening values (8 p.m.) were significantly lower than morning values, but in migraineurs the decrease of the three peptides was less. The raised proopiocortin-related peptide plasma levels found in children suffering from migraine cannot be explained at present, although the discomfort experienced by the patients may create a situation of chronic stress which could explain such a finding. Whatever the explanation is, these findings differentiate prepubertal migraine from the most severe forms of headache occurring in adult life where lower than normal opioid levels have been demonstrated.


Ribosomal protein S14 negatively regulates c-Myc activity.
The ribosomal gene RPS14 is associated with the cancer-prone 5q-syndrome, which is caused by an interstitial deletion of the long arm of human chromosome 5. Previously, we found that ribosomal protein S14 (RPS14) binds to and inactivates MDM2, consequently leading to p53-dependent cell-cycle arrest and growth inhibition. However, it remains elusive whether RPS14 regulates cell proliferation in a p53-independent manner. Here, we show that RPS14 interacts with the Myc homology box II (MBII) and the C-terminal basic helix-loop-helix leucine zipper (bHLH-LZ) domains of the oncoprotein c-Myc. Further, RPS14 inhibited c-Myc transcriptional activity by preventing the recruitment of c-Myc and its cofactor, TRRAP, to the target gene promoters, as thus suppressing c-Myc-induced cell proliferation. Also, siRNA-mediated RPS14 depletion elevated c-Myc transcriptional activity determined by its target gene, Nucleolin, expression. Interestingly, RPS14 depletion also resulted in the induction of c-Myc mRNA and subsequent protein levels. Consistent with this, RPS14 promoted c-Myc mRNA turnover through an Argonaute 2 (Ago2)- and microRNA-mediated pathway. Taken together, our study demonstrates that RPS14 negates c-Myc functions by directly inhibiting its transcriptional activity and mediating its mRNA degradation via miRNA.


HIV vaccine rationale, design and testing.
A central obstacle to the design of a global HIV vaccine is viral diversity. Antigenic differences in envelope proteins result in distinct HIV serotypes, operationally defined such that antibodies raised against envelope molecules from one serotype will not bind envelope molecules from a different serotype. The existence of serotypes has presented a similar challenge to vaccine development against other pathogens. In such cases, antigenic diversity has been addressed by vaccine design. For example, the poliovirus vaccine includes three serotypes of poliovirus, and Pneumovax presents a cocktail of 23 pneumococcal variants to the immune system. It is likely that a successful vaccine for HIV must also comprise a cocktail of antigens. Here, data relevant to the development of cocktail vaccines, designed to harness diverse, envelope-specific B-cell and T-cell responses, are reviewed.


Preventive health risk appraisal for older people and impact on GPs' patient management: a prospective study.
Health risk appraisals (HRAs) are recommended for detection of potentially modifiable risk factors for health status decline of older people. Little is known how family physicians manage detected risk factors. We evaluated (i) if risk factors in one or more of five predefined domains were detected in a primary care-based HRA and (ii) how often these findings had an impact on the further management of patients. We performed a prospective observational study in a rural community in Austria and included persons (age >or= 70 years) living at home. We applied the standardized assessment for elderly people in primary care (STEP) instrument and evaluated risk factors for status decline assessing five domains (cognitive function, depression, urinary incontinence, hearing impairment and mobility/falls). Two hundred and sixty-four persons participated and the HRA revealed a wide range of risk factors for health status decline [from 4.5% (12/264) in the depression domain up to 31% (81/264) for mobility/falls and 41% (107/264) in the cognitive domain]. The findings had an impact on the further management in four domains: hearing impairment (100% of findings with impact), mobility/falls (93%), depression (83%) and urinary incontinence (65%). In contrast, abnormal cognitive findings lead to action only in every fifth participant (18%; 19/107). In contrast to other domains, family physicians are hesitant to act upon abnormal findings of cognitive testing. Additional knowledge is needed to clarify the value of abnormal cognitive findings for management of patients and support of their carers.


Candidate Enzymes for Saffron Crocin Biosynthesis Are Localized in Multiple Cellular Compartments.
Saffron is the dried stigmas of Crocus sativus and is the most expensive spice in the world. Its red color is due to crocins, which are apocarotenoid glycosides that accumulate in the vacuole to a level up to 10% of the stigma dry weight. Previously, we characterized the first dedicated enzyme in the crocin biosynthetic pathway, carotenoid cleavage dioxygenase2 (CsCCD2), which cleaves zeaxanthin to yield crocetin dialdehyde. In this work, we identified six putative aldehyde dehydrogenase (ALDH) genes expressed in C. sativus stigmas. Heterologous expression in Escherichia coli showed that only one of corresponding proteins (CsALDH3I1) was able to convert crocetin dialdehyde into the crocin precursor crocetin. CsALDH3I1 carries a carboxyl-terminal hydrophobic domain, similar to that of the Neurospora crassa membrane-associated apocarotenoid dehydrogenase YLO-1. We also characterized the UDP-glycosyltransferase CsUGT74AD1, which converts crocetin to crocins 1 and 2'. In vitro assays revealed high specificity of CsALDH3I1 for crocetin dialdehyde and long-chain apocarotenals and of CsUGT74AD1 for crocetin. Following extract fractionation, CsCCD2, CsALDH3I1, and CsUGT74AD1 were found in the insoluble fraction, suggesting their association with membranes or large insoluble complexes. Analysis of protein localization in both C. sativus stigmas and following transgene expression in Nicotiana benthamiana leaves revealed that CsCCD2, CsALDH3I, and CsUGT74AD1 were localized to the plastids, the endoplasmic reticulum, and the cytoplasm, respectively, in association with cytoskeleton-like structures. Based on these findings and current literature, we propose that the endoplasmic reticulum and cytoplasm function as transit centers for metabolites whose biosynthesis starts in the plastid and are accumulated in the vacuole.


The translational machinery is an optimized molecular network that affects cellular homoeostasis and disease.
Translation involves interactions between mRNAs, ribosomes, tRNAs and a host of translation factors. Emerging evidence on the eukaryotic translational machinery indicates that these factors are organized in a highly optimized network, in which the levels of the different factors are finely matched to each other. This optimal factor network is essential for producing proteomes that result in optimal fitness, and perturbations to the optimal network that significantly affect translational activity therefore result in non-optimal proteomes, fitness losses and disease. On the other hand, experimental evidence indicates that translation and cell growth are relatively robust to perturbations, and viability can be maintained even upon significant damage to individual translation factors. How the eukaryotic translational machinery is optimized, and how it can maintain optimization in the face of changing internal parameters, are open questions relevant to the interaction between translation and cellular disease states.


Advances of phytotherapy in ischemic stroke targeting PI3K/Akt signaling.
The pathogenesis of ischemic stroke is complex, and PI3K/Akt signaling is considered to play a crucial role in it. The PI3K/Akt pathway regulates inflammation, oxidative stress, apoptosis, autophagy, and vascular endothelial homeostasis after cerebral ischemia; therefore, drug research targeting the PI3K/Akt pathway has become the focus of scientists. In this review, we analyzed the research reports of antiischemic stroke drugs targeting the PI3K/Akt pathway in the past two decades. Because of the rich sources of natural products, increasing studies have explored the value of natural compounds, including Flavonoids, Quinones, Alkaloids, Phenylpropanoids, Phenols, Saponins, and Terpenoids, in alleviating neurological impairment and achieved satisfactory results. Herbal extracts and medicinal formulas have been applied in the treatment of ischemic stroke for thousands of years in East Asian countries. These precious clinical experiences provide a new avenue for research of antiischemic stroke drugs. Finally, we summarize and discuss the characteristics and shortcomings of the current research and put forward prospects for further in-depth exploration.


Structure and origin of the acute promyelocytic leukemia myl/RAR alpha cDNA and characterization of its retinoid-binding and transactivation properties.
Acute promyelocytic leukemia (APL) is characterized by the 15;17 chromosomal translocation. Cloning experiments have established that the chromosome 17 breakpoint maps to the RAR alpha and the 15 to the myl locus. The resulting chimeric gene is transcribed as a myl/RAR alpha fusion mRNA. By isolating both normal myl and APL myl/RAR alpha cDNAs, we showed that the myl/RAR alpha mRNA encodes for a putative fusion protein with a molecular weight of about 103 kDa, which is made up of 530 amino acids derived from the myl N-terminus and 402 amino acids originating from the RAR alpha C-terminus. The protein includes the RAR alpha DNA and retinoid-binding regions but lacks the A portion of the N-terminal region (A/B region) which is thought to contain one of the RAR alpha transactivation domains. The myl/RAR alpha protein acted as a retinoid-inducible transcription factor with both ligand-independent repressor and ligand-dependent activator functions in transactivation experiments of a retinoic acid-responsive gene. Myl/RAR alpha exerted this dual function three times more effectively than RAR alpha and had about 10-fold greater affinity for RA than RAR alpha. Comparison of myl/RAR alpha genomic and cDNA sequences from the same case demonstrated that both chromosome 15 and 17 breakpoints occurred within introns and the myl and RAR alpha sequences are spliced in the same polyadenylated RNA.


Enhanced expression of glucose-6-phosphate dehydrogenase in human cells sustaining oxidative stress.
Recent reports have demonstrated that glucose-6-phosphate dehydrogenase (G6PD) activity in mammalian cells is necessary in order to ensure cell survival when damage is produced by reactive oxygen intermediates. In this paper we demonstrate that oxidative stress, caused by agents acting at different steps in the biochemical pathway controlling the intracellular redox status, determines the increase in G6PD-specific activity in human cell lines of different tissue origins. The intracellular level of G6PD-specific mRNA also increases, with kinetics compatible with the induction of new enzyme synthesis. We carried out experiments in which cells were exposed to oxidative stress in the presence of inhibitors of protein or RNA synthesis. These demonstrated that increased G6PD expression is mainly due to an increased rate of transcription, with a minor but significant contribution of regulatory mechanisms acting at post-transcriptional levels. These results provide new information on the defence systems that eukaryotic cells possess in order to prevent damage caused by potentially harmful oxygen derivatives.


Seminested PCR for detection and identification of Candida species directly from blood culture bottles.
We investigated the performance of a seminested PCR (snPCR) assay carried out directly from overnight incubated blood culture bottles of 50 newborn intensive care unit (NICU) patients with suspected candidemia and compared these, for sensitivity, specificity and reliability with results from blood cultures. All positive blood cultures (n = 17) yielded positive results for snPCR, which detected the same Candida species, as did the yeast isolates of which 13 were C. parapsilosis and 4 were C. albicans. With both assays showing 32 negative samples and one sample positive with snPCR but negative with blood culture, sensitivity and specificity of snPCR were 100% and 97%, respectively. The patient with contradictory results exhibited a positive blood culture one week later yielding the same species as identified by snPCR. These are the first data demonstrating that snPCR from overnight blood culture bottles can be a potential tool for rapid detection and identification of Candida species, allowing follow-up of the "gold standard" blood culturing, as well.


Glycosylated superparamagnetic nanoparticle gradients for osteochondral tissue engineering.
In developmental biology, gradients of bioactive signals direct the formation of structural transitions in tissue that are key to physiological function. Failure to reproduce these native features in an in vitro setting can severely limit the success of bioengineered tissue constructs. In this report, we introduce a facile and rapid platform that uses magnetic field alignment of glycosylated superparamagnetic iron oxide nanoparticles, pre-loaded with growth factors, to pattern biochemical gradients into a range of biomaterial systems. Gradients of bone morphogenetic protein 2 in agarose hydrogels were used to spatially direct the osteogenesis of human mesenchymal stem cells and generate robust osteochondral tissue constructs exhibiting a clear mineral transition from bone to cartilage. Interestingly, the smooth gradients in growth factor concentration gave rise to biologically-relevant, emergent structural features, including a tidemark transition demarcating mineralized and non-mineralized tissue and an osteochondral interface rich in hypertrophic chondrocytes. This platform technology offers great versatility and provides an exciting new opportunity for overcoming a range of interfacial tissue engineering challenges.


Alkali metal atom-aromatic ring: a novel interaction mode realizes large first hyperpolarizabilities of M@AR (M=Li, Na, and K, AR=pyrrole, indole, thiophene, and benzene).
Several new electride compounds M@pyrrole (M = Li, Na, and K), Li@AR (AR = indole, thiophene, and benzene), Li@tryptophan and Li@serotonin were designed and investigated, which exhibit considerably large first hyperpolarizabilities (β(0)) (6705, 1116, 11399, 5781, 4808, 1536, 8106, and 9389 au, respectively) by comparison with their corresponding sole molecules pyrrole (β(0) = 30 au), indole (104 au), thiophene (6 au), benzene (0 au), tryptophan (159 au) and serotonin (151 au), respectively. The computational results revealed that the interaction of the alkali metal atom with π-conjugated aromatic ring (AR) is one effectively new approach to produce diffuse excess electron to get a large β(0) value, which is advantageous for the design of the novel high-performance NLO materials with π-conjugated AR: alkali metal atoms doped nanomaterials and biomolecules.


Nonlinear vocal phenomena affect human perceptions of distress, size and dominance in puppy whines.
While nonlinear phenomena (NLP) are widely reported in animal vocalizations, often causing perceptual harshness and roughness, their communicative function remains debated. Several hypotheses have been put forward: attention-grabbing, communication of distress, exaggeration of body size and dominance. Here, we use state-of-the-art sound synthesis to investigate how NLP affect the perception of puppy whines by human listeners. Listeners assessed the distress, size or dominance conveyed by synthetic puppy whines with manipulated NLP, including frequency jumps and varying proportions of subharmonics, sidebands and deterministic chaos. We found that the presence of chaos increased the puppy's perceived level of distress and that this effect held across a range of representative fundamental frequency (fo) levels. Adding sidebands and subharmonics also increased perceived distress among listeners who have extensive caregiving experience with pre-weaned puppies (e.g. breeders, veterinarians). Finally, we found that whines with added chaos, subharmonics or sidebands were associated with larger and more dominant puppies, although these biases were attenuated in experienced caregivers. Together, our results show that nonlinear phenomena in puppy whines can convey rich information to human listeners and therefore may be crucial for offspring survival during breeding of a domesticated species.


Challenges in managing patients referred for eosinophilic esophagitis: A telephone survey and retrospective review.
Background: Eosinophilic esophagitis (EoE) in patients referred to allergists can be difficult to manage. This is due to multiple factors, including overlaps in presentation with gastroesophageal reflux disease and proton-pump inhibitor responsive eosinophilia, remaining uncertainties regarding the role of different forms of allergy testing, and a variety of patient adherence issues. Objective: To highlight, in an objectively studied fashion, complications that can be faced when managing patients referred for EoE. Methods: We conducted a telephone survey of 78 patients (pediatric and adult) who were referred to our academic allergy clinic for EoE. The survey focused on patients' perspectives regarding their symptoms and treatments. We then conducted a chart review to determine if there had been a proton-pump inhibitor (PPI) trial before diagnosis, and we compared patient responses with documented allergy test results, treatment plans, and biopsy results. Results: Only 22 of 78 patients (28%) had a ≥8-week PPI trial before diagnosis and/or referral. There was considerable variability in the type of allergy testing done for patients, and how the results were used to guide therapy. More than one-third of the patients reported being on a different treatment regimen (PPI, swallowed steroid, and/or diet) than planned, and the majority of patients on dietary therapy reported being on a different diet than planned. Also, nearly half of the adult patients did not have follow-up biopsies done despite recommendations for this. Conclusion: We identified several challenges in EoE management, including potential misdiagnosis or overtreatment, lack of standardization in testing and dietary recommendations, and patient adherence issues. We hope this information will prompt increased vigilance for these issues and promote solutions when needed.


Molecular forms, binding functions, and developmental expression patterns of cytotactin and cytotactin-binding proteoglycan, an interactive pair of extracellular matrix molecules.
Cytotactin is an extracellular matrix protein that is found in a restricted distribution and is related to developmental patterning at a number of neural and non-neural sites. It has been shown to bind specifically to other extracellular matrix components including a chondroitin sulfate proteoglycan (cytotactin-binding [CTB] proteoglycan) and fibronectin. Cell binding experiments have revealed that cytotactin interacts with neurons and fibroblasts. When isolated from brain, both cytotactin and CTB proteoglycan contain the HNK-1 carbohydrate epitope. Here, specific antibodies prepared against highly purified cytotactin and CTB proteoglycan were used to correlate the biochemical alterations and modes of binding of these proteins with their differential tissue expression as a function of time and place during chicken embryo development. It was found that, during neural development, both the levels of expression of cytotactin and CTB proteoglycan and of the molecular forms of each molecule varied, following different time courses. In addition, a novel Mr 250,000 form of cytotactin was detected that contained chondroitin sulfate. The intermolecular binding of cytotactin and CTB proteoglycan and the binding of cytotactin to fibroblasts were characterized further and found to be inhibited by EDTA, consistent with a dependence on divalent cations. Unlike the molecules from neural tissue, cytotactin and CTB proteoglycan isolated from non-neural tissues such as fibroblasts lacked the HNK-1 epitope. Nevertheless, the intermolecular and cellular binding activities of cytotactin isolated from fibroblast culture medium were comparable to those of the molecule isolated from brain, suggesting that the HNK-1 epitope is not directly involved in binding. Binding experiments involving enzymatically altered molecules that lack chondroitin sulfate suggested that this glycosaminoglycan is also not directly involved in binding. Although they clearly formed a binding couple, the spatial distributions of cytotactin and CTB proteoglycan in the embryo were not always coincident. They were similar in tissue sections from the cerebellum, gizzard, and vascular smooth muscle. In contrast, CTB proteoglycan was present in cardiac muscle where no cytotactin is present, and it was seen in cartilage throughout development unlike cytotactin, which was present only in immature chondrocytes. Cell culture experiments were consistent with the previous conclusion that cytotactin was specifically synthesized by glia, whereas CTB proteoglycan was specifically synthesized by neurons.(ABSTRACT TRUNCATED AT 400 WORDS)


Development of a Nicotinic Acetylcholine Receptor nAChR α7 Binding Activity Prediction Model.
Despite the well-known adverse health effects associated with tobacco use, addiction to nicotine found in tobacco products causes difficulty in quitting among users. Nicotinic acetylcholine receptors (nAChRs) are the physiological targets of nicotine and facilitate addiction to tobacco products. The nAChR-α7 subtype plays an important role in addiction; therefore, predicting the binding activity of tobacco constituents to nAChR-α7 is an important component for assessing addictive potential of tobacco constituents. We developed an α7 binding activity prediction model based on a large training data set of 843 chemicals with human α7 binding activity data extracted from PubChem and ChEMBL. The model was tested using 1215 chemicals with rat α7 binding activity data from the same databases. Based on the competitive docking results, the docking scores were partitioned to the key residues that play important roles in the receptor-ligand binding. A decision forest was used to train the human α7 binding activity prediction model based on the partition of docking scores. Five-fold cross validations were conducted to estimate the performance of the decision forest models. The developed model was used to predict the potential human α7 binding activity for 5275 tobacco constituents. The human α7 binding activity data for 84 of the 5275 tobacco constituents were experimentally measured to confirm and empirically validate the prediction results. The prediction accuracy, sensitivity, and specificity were 64.3, 40.0, and 81.6%, respectively. The developed prediction model of human α7 may be a useful tool for high-throughput screening of potential addictive tobacco constituents.


Diagnostic accuracy and recall rates for digital mammography and digital mammography combined with one-view and two-view tomosynthesis: results of an enriched reader study.
The purpose of this study was to compare two methods of combining tomosynthesis with digital mammography by assessing diagnostic accuracy and recall rates for digital mammography alone and digital mammography combined with one-view tomosynthesis and two-view tomosynthesis. Three hundred ten cases including biopsy-proven malignancies (51), biopsy-proven benign findings (47), recalled screening cases (138), and negative screening cases (74) were reviewed by 15 radiologists sequentially using digital mammography, adding one-view tomosynthesis, and then two-view tomosynthesis. Cases were assessed for recall and assigned a BI-RADS score and probability of malignancy for each imaging method. Diagnostic accuracy was assessed using receiver operating characteristic (ROC) analysis. Screening recall rates were compared using pooled logistical regression analysis. A p value of < 0.0167 was considered significant. The area under the ROC curve (AUC) for digital mammography (DM), DM plus one-view tomosynthesis, and DM plus two-view tomosynthesis was 0.828, 0.864, and 0.895, respectively. Both one-view and two-view tomosynthesis plus DM were significantly better than DM alone (Δ AUCs 0.036 [p = 0.009] and 0.068 [p < 0.001]). Average noncancer recall rates for digital mammography, DM plus one-view tomosynthesis, and DM plus two-view tomosynthesis were 44.2%, 27.2%, and 24.0%, respectively. Combined with DM, one-view and two-view tomosynthesis both showed significantly lower noncancer recall rates than digital mammography alone (p < 0.001). Digital mammography with two-view tomosynthesis showed a significantly lower recall rate than digital mammography with one-view tomosynthesis (p < 0.001). Diagnostic accuracy for dense (Δ AUC, 0.091%; p < 0.001) and nondense (Δ AUC, 0.035%; p = 0.001) breasts improved with DM plus two-view tomosynthesis compared with digital mammography alone. Compared with digital mammography, diagnostic sensitivity for invasive cancers increased with the addition of both one-view (Δ12.0%, p < 0.001) and two-view (Δ21.7%, p < 0.001) tomosynthesis. The addition of one-view tomosynthesis to conventional digital mammography improved diagnostic accuracy and reduced the recall rate; however, the addition of two-view tomosynthesis provided twice the performance gain in diagnostic accuracy while further reducing the recall rate.


Human leukocyte antigen (HLA) class II peptide flanking residues tune the immunogenicity of a human tumor-derived epitope.
CD4+ T-cells recognize peptide antigens, in the context of human leukocyte antigen (HLA) class II molecules (HLA-II), which through peptide-flanking residues (PFRs) can extend beyond the limits of the HLA binding. The role of the PFRs during antigen recognition is not fully understood; however, recent studies have indicated that these regions can influence T-cell receptor (TCR) affinity and pHLA-II stability. Here, using various biochemical approaches including peptide sensitivity ELISA and ELISpot assays, peptide-binding assays and HLA-II tetramer staining, we focused on CD4+ T-cell responses against a tumor antigen, 5T4 oncofetal trophoblast glycoprotein (5T4), which have been associated with improved control of colorectal cancer. Despite their weak TCR-binding affinity, we found that anti-5T4 CD4+ T-cells are polyfunctional and that their PFRs are essential for TCR recognition of the core bound nonamer. The high-resolution (1.95 Å) crystal structure of HLA-DR1 presenting the immunodominant 20-mer peptide 5T4111-130, combined with molecular dynamic simulations, revealed how PFRs explore the HLA-proximal space to contribute to antigen reactivity. These findings advance our understanding of what constitutes an HLA-II epitope and indicate that PFRs can tune weak affinity TCR-pHLA-II interactions.


An evaluation of mechanical property and microstructural development in HAP-Ca polycarboxylate biocomposites prepared by hot pressing.
A hot-pressing technique was used to prepare composites anticipated to be biocompatible. Ca(4)(PO(4))(2)O (TetCP) was reacted with an acrylic-itaconic copolymer (CoP) in the absence of a solvent to form composites comprised of Ca(10)(PO(4))(6)(OH)(2') (hydroxyapatite, or HAp) and the Ca polyalkenoate salt. The effect of temperature, pressure, and hot-pressing time on the mechanical properties and microstructure of the composites were studied. Results showed that both tensile strength and elastic modulus increased when temperature and time were increased. When the compaction pressure was increased, these properties initially increased but decreased at high pressures. These variations in the mechanical properties were correlated with the microstructure of these composites. The mechanism of the reaction was also studied. Reaction starts when the copolymer is heated to above its T(g) permitting it to flow and react with the TetCP grains. The COOH groups on the polymer are neutralized by Ca(2+) ions liberated from the TetCP. At the end of reaction, a network of the Ca polyalkenoate salt is formed in which HAp crystals are embedded.


Factors and Conditions That Impact Electroporation of Clostridioides difficile Strains.
An important risk factor for acquiring Clostridioides difficile infection is antibiotic use. Therefore, a detailed knowledge of the physiology and the virulence factors can help drive the development of new diagnostic tools and nonantibiotic therapeutic agents to combat these organisms. Several genetic systems are available to study C. difficile in the laboratory environment, and all rely on stably replicating or segregationally unstable plasmids. Currently, the transfer of plasmids into C. difficile can only be performed by conjugation using Escherichia coli or Bacillus subtilis as conjugal donors. Here we report a method to introduce plasmid DNA into C. difficile using electroporation and test factors that might contribute to higher transformation efficiencies: osmolyte used to stabilize weakened cells, DNA concentration, and recovery time postelectroporation. Depending on the C. difficile strain and plasmid used, this transformation protocol achieves between 20 and 200 colonies per microgram of DNA and is mostly influenced by the recovery time postelectroporation. Based on our findings, we recommend that each strain be tested for the optimum recovery time in each lab.IMPORTANCE Understanding the underlying biology of pathogens is essential to develop novel treatment options. To drive this understanding, genetic tools are essential. In recent years, the genetic toolbox available to Clostridioides difficile researchers has expanded significantly but still requires the conjugal transfer of DNA from a donor strain into C. difficile Here we describe an electroporation-based transformation protocol that was effective at introducing existing genetic tools into different C. difficile strains.


[Fever of unknown origin: differences in two different clinical series in a universitary hospital].
We compare two clinical series of patients with fever of unknown origin (FUO) during two different periods of time (1982-1989 and 1997-2004) in the department of Internal Medicine at La Paz University Hospital, Madrid. We have analysed the changes in aetiology and diagnostic methods in these separated periods. We describe a decrease in the incidence of this clinical entity which we think to be due to the improvement of the diagnostic methods. This provides an earlier diagnosis and so patients do not reach criteria for classical FUO. Due to these improvements we have also assessed a change in the final diagnoses and the means used to reach them which, in general, have been less interventionist.


Expression of microRNAs and its regulation in plants.
MicroRNAs (miRNAs) have emerged as an essential regulatory component in plants. Many of the known miRNAs are evolutionarily conserved across diverse plant species and function in the regulatory control of fundamentally important biological processes such as developmental timing, patterning, and response to environmental changes. Expression of miRNAs in plants involves transcription from MIRNA loci by RNA polymerase II (pol II), multi-step processing of the primary transcripts by the DICER-LIKE1 (DCL1) complex, and formation of effector complexes consisting of mature miRNAs and ARGONAUTE (AGO) family proteins. In this short review, we present the most recent advances in our understanding of the molecular machinery as well as the regulatory mechanisms involved in the expression of miRNAs in plants.


The 10th Japan Bioanalysis Forum Symposium conference: a report.
The Japan Bioanalysis Forum Symposium was held on 12-14 February 2019 (Yokohama, Japan), in celebration of its 10th anniversary, and over 370 participants from pharmaceutical industries, contractors, academia and regulatory authorities from home and abroad came together in Yokohama. The 3-day symposium particularly aimed to foster collaboration with the scientists surrounding bioanalysts, according to the theme 'Open to the Public.' The symposium also included a broad range of pioneering programs, such as lectures by speakers from DMPK/metabolomics fields, discussions of future bioanalysis and poster presentations by publicly offered presenters as well as the regular ones we had organized. This report summarizes the major topics as a conference report.


Shedding of c-Met is regulated by crosstalk between a G-protein coupled receptor and the EGF receptor and is mediated by a TIMP-3 sensitive metalloproteinase.
A wide repertoire of transmembrane proteins are proteolytically released from the cell surface by a process known as 'ectodomain shedding', under both normal and pathophysiological conditions. Little is known about the physiological mechanisms that regulate this process. As a model system, we have investigated the metalloproteinase-mediated cleavage of the hepatocyte growth factor receptor, Met. We show that epidermal growth factor (EGF) receptor activation, either directly by EGF or indirectly via the G-protein coupled receptor (GPCR) agonist lysophosphatidic acid (LPA), induces cleavage of Met through activation of the Erk MAP kinase signalling cascade. The tyrosine kinase activity of the EGFR was a prerequisite for this stimulation, since treatment of cells with a synthetic inhibitor of this receptor, AG1478, completely abrogated shedding. The metalloproteinase mediating Met cleavage was specifically inhibited by the tissue inhibitor of metalloproteinases (TIMP)-3, but not by TIMP-1 or TIMP-2. Furthermore, the level of Met shedding could be modulated by different cell-matrix interactions. Our results indicate that ectodomain shedding is a highly regulated process that can be stimulated by EGFR signalling pathways and integrin ligation.


Role of microglia in central nervous system infections.
The nature of microglia fascinated many prominent researchers in the 19th and early 20th centuries, and in a classic treatise in 1932, Pio del Rio-Hortega formulated a number of concepts regarding the function of these resident macrophages of the brain parenchyma that remain relevant to this day. However, a renaissance of interest in microglia occurred toward the end of the 20th century, fueled by the recognition of their role in neuropathogenesis of infectious agents, such as human immunodeficiency virus type 1, and by what appears to be their participation in other neurodegenerative and neuroinflammatory disorders. During the same period, insights into the physiological and pathological properties of microglia were gained from in vivo and in vitro studies of neurotropic viruses, bacteria, fungi, parasites, and prions, which are reviewed in this article. New concepts that have emerged from these studies include the importance of cytokines and chemokines produced by activated microglia in neurodegenerative and neuroprotective processes and the elegant but astonishingly complex interactions between microglia, astrocytes, lymphocytes, and neurons that underlie these processes. It is proposed that an enhanced understanding of microglia will yield improved therapies of central nervous system infections, since such therapies are, by and large, sorely needed.


Visual prognosis and vitreous molecules after vitrectomy for macular edema with branch retinal vein occlusion.
This study investigated whether vascular endothelial growth factor (VEGF), soluble intercellular adhesion molecule-1 (sICAM-1), and pigment epithelium-derived factor (PEDF) influence the visual prognosis of patients with macular edema and branch retinal vein occlusion (BRVO). In 47 consecutive patients (47 eyes) undergoing vitrectomy, retinal thickness was examined by optical coherence tomography. Best-corrected visual acuity and the vitreous fluid levels of VEGF, sICAM-1, and PEDF were also determined. Patients were followed for at least 6 months after surgery. Vitreous fluid levels of VEGF and sICAM-1 were significantly lower in the patients with more marked improvement of visual acuity after vitrectomy, while PEDF was significantly higher. VEGF and sICAM-1 levels were significantly higher in patients with greater postoperative improvement of macular edema, while PEDF was significantly lower. In BRVO patients, vitreous fluid levels of VEGF, sICAM-1, and PEDF may influence both the response of macular edema to vitrectomy and the visual prognosis.


Theoretical characterization of ion channel blockade: ligand binding to periodically accessible receptors.
With repetitive stimulation, the time course of use-dependent blockade as assessed by peak membrane ion currents can be described by a sequence of blocking relationships that have the form of recurrence equations. The equations of the sequence describe blockade acquired during each interval of a stimulus where the possibly different binding and unbinding rates are assumed constant during each interval. The solution predicts that use-dependent uptake follows an exponential time course. Furthermore, the exponential uptake rate is a linear function of uptake rates associated with the stimulus time intervals. Similarly, the fraction of blocked channels at steady state is a linear function of the interval dependent blockade equilibria. Several novel tests of consistency between the model and observations are derived from these theoretical results. It is also shown that as the stimulus interval increases to infinity, steady state dissociation constants measured by peak membrane currents are theoretically equivalent to those measured with true equilibrium methods such as radioligand binding studies.


Nm23: an invasion suppressor gene in CNS tumours?
The nm23 gene family is widely documented as a suppressor of the metastatic phenotype in many human cancers. However, its role with respect to the invasive behavior of primary central nervous system tumours has scarcely been addressed. This review describes the members of this family identified to date and outlines the in vitro and in vivo data gathered from nearly ten years of nm23 research within the context of malignant progression in peripheral tumours. It is clear that this is an area which can no longer be ignored by the brain tumor community. Human nm23 bears strong sequence homology to the awd gene in Drosophila, mutations in which cause abnormal morphology of larval neural tissue. The functions and possible mechanisms underlying the ability of nm23 to suppress metastatic behavior are discussed.


[Lifestyle and global cardiovascular risk: a prospective study on a borderline hypertensive population].
Arterial hypertension is a well known risk factor for cardiovascular diseases. Today, it is possible to calculate the cardiovascular risk at 10 years with the risk card. The reduction of cardiovascular risk is based on a multi-factorial approach including the lifestyle modification. In Italy, OEC study has calculated that a certain proportion of borderline hypertensives, not eligible for a pharmacological treatment, remain at risk. Borderline arterial hypertension (140-150/90-95 mmHg) in Italian population is documented in 19% of males and 14% of females.: Aim of the study is to examine the efficacy of the lifestyle changes in reducing the global cardiovascular risk in bordeline hypertensives. 102 patients affected by borderline hypertensive (46 M and 56 F, mean age: 55.6 yrs ) were enclosed in a 12 months prospective study. Three checks were programmed during the follow-up, i.e., at beginning, 6 months and 12 months later. At the start of the study every patient received a list of lifestyle changes to be respected. Pressure arterial systolic and diastolic were obtained at beginning and at the end of successive. At the last check each patient received a questionnaire to be filled up. According to the calculated score, each patient was classified as: non-responder (score: 0-3), partially responder (score: 4-6), responder (score: 7-9). A significant reduction of the globalcardiovascular risk has been observed at the end of the study in both the responders and partially responders. Such a reduction was seen to be due to positive changes in blood pressure and total, HDL, LDL cholesterol. This study confirmed that a non-pharmacological therapy based on lifestyle changes has to be preventively administered in the presence of a borderline hypertension because of its beneficial effects in reducing the global risk of cardiovascular disease. Therefore, we firmly think that a prompt utilization of a correct lifestyle can sort the triple effect of improving the expectancy of life, ameliorating the quality of life, reducing the social costs of arterial hypertension.


Diagnosis clusters for emergency medicine.
Aggregated emergency department (ED) data are useful for research, ED operations, and public health surveillance. Diagnosis data are widely available as The International Classification of Diseases, version, 9, Clinical Modification (ICD-9-CM) codes; however, there are over 24,000 ICD-9-CM code-descriptor pairs. Standardized groupings (clusters) of ICD-9-CM codes have been developed by other disciplines, including family medicine (FM), internal medicine (IM), inpatient care (Agency for Healthcare Research and Quality [AHRQ]), and vital statistics (NCHS). The purpose of this study was to evaluate the coverage of four existing ICD-9-CM cluster systems for emergency medicine. In this descriptive study, four cluster systems were used to group ICD-9-CM final diagnosis data from a southeastern university tertiary referral center. Included were diagnoses for all ED visits in July 2000 and January 2001. In the comparative analysis, the authors determined the coverage in the four cluster systems, defined as the proportion of final diagnosis codes that were placed into clusters and the frequencies of diagnosis codes in each cluster. The final sample included 7,543 visits with 19,530 diagnoses. Coverage of the ICD-9-CM codes in the ED sample was: AHRQ, 99%; NCHS, 88%; FM, 71%; IM, 68%. Seventy-six percent of the AHRQ clusters were small, defined as grouping <1% of the diagnosis codes in the sample. The AHRQ system provided the best coverage of ED ICD-9-CM codes. However, most of the clusters were small and not significantly different from the raw data.


Comparison of hydrology of wetlands in Pennsylvania and Oregon (USA) as an indicator of transferability of hydrogeomorphic (HGM) functional models between regions.
The hydrogeomorphic (HGM) approach to wetland classification and functional assessment is becoming more widespread in the United States but its use has been limited by the length of time needed to develop appropriate data sets and functional assessment models. One particularly difficult aspect is the transferability among geographic regions of specific models used to assess wetland function. Sharing of models could considerably shorten development and implementation of HGM throughout the United States and elsewhere. As hydrology is the driving force behind wetland functions, we assessed the comparability of hydrologic characteristics of three HGM subclasses (slope, headwater floodplain, mainstem floodplain) using comparable long-term hydrologic data sets from different regions of the United States (Ridge and Valley Province in Pennsylvania and the Willamette Valley in Oregon). If hydrology by HGM subclass were similar between different geographic regions, it might be possible to more readily transfer extant models between those regions. We found that slope wetlands (typically groundwater-driven) had similar hydrologic characteristics, even though absolute details (such as depth of water) differed. We did not find the floodplain subclasses to be comparable, likely due to effects of urbanization in Oregon, regional differences in soils and, perhaps, climate. Slight differences in hydrology can shift wetland functions from those mediated by aerobic processes to those dominated by anaerobic processes. Functions such as nutrient cycling can be noticeably altered as a result. Our data suggest considerable caution in the application of models outside of the region for which they were developed.


Social support for exercise by experts in older women post-hip fracture.
Using the data of the Baltimore Hip Study 5 (a home-based exercise intervention), this study examined how social support for exercise by experts (SSE-E) affected the self-efficacy, outcome expectations, and exercise behavior among older women following a hip fracture. The total sample included 164 females aged 65 years (M = 81.0; SD = 6.9) who had surgical repair of a nonpathologic hip fracture. Model testing showed a direct relationship between SSE-E and outcome expectations for exercise. There was, however, no direct or indirect relationship between SSE-E and self-efficacy or exercise behavior. The positive effect of SSE-E on the outcome expectations for exercise in older women recovering from a hip fracture provides an opportunity for health care providers in improving physical activity in this population.


Analysis of sedimentation equilibrium distributions reflecting nonideal macromolecular associations.
A rigorous statistical-mechanical approach is adopted to derive general quantitative expressions that allow for the effects of thermodynamic nonideality in equilibrium measurements reflecting interaction between dissimilar macromolecular reactants. An analytical procedure based on these expressions is then formulated for obtaining global estimates of equilibrium constants and the corresponding reference thermodynamic activities of the free reactants in each of several sedimentation equilibrium experiments. The method is demonstrated by application to results from an ultracentrifugal study of an electrostatic interaction between ovalbumin and cytochrome c (Winzor, D. J., M. P. Jacobsen, and P. R. Wills. 1998. Biochemistry. 37:2226-2233). It is demonstrated that reliable estimates of relevant thermodynamic parameters are extracted from the data through statistical analysis by means of a simple nonlinear fitting procedure.


Effect of fludarabine and arabinosylcytosine on multidrug resistant cells.
Anthracyclines are first-line drugs in the treatment of acute leukemia, but the sensitivity of leukemic cells to anthracyclines can be downmodulated by multidrug resistance (MDR) transport proteins like Pgp. Pgp overexpression is negatively related to treatment response. Alternative drugs may be required to overcome the MDR problem. Arabinosylcytosine (ara-C) and 9-beta-D-arabinofuranosyl-2-fluoro-adenine monophosphate (fludarabine, F-ara) were tested alone and in combination in four pairs of leukemia and tumor non-MDR and MDR cell lines. Toxicity was assayed by growth inhibition with the microcultured MTT assay. MDR cells were more sensitive than or as sensitive as non-MDR cells to ara-C and to F-ara alone. The resistance index to ara-C was decreased upon pre-exposure of the MDR cells to low-dose F-ara (10 ng/mL), showing that the combination of ara-C and F-ara was more active on MDR cells than on non-MDR parental ones. Neither sensitivity to ara-C nor sensitivity to F-ara was influenced by Pgp overexpression. These data provide a rationale for more extensive and more intensive testing of combinations of ara-C and F-ara in Pgp-mediated MDR acute leukemia. In relapsed/resistant and in secondary acute leukemias, increasing the dose of ara-C and combining ara-C with F-ara might be more rewarding than administering anthracyclines or other Pgp-processable compounds.


Disease- and Therapy-Specific Impact on Humoral Immune Responses to COVID-19 Vaccination in Hematologic Malignancies.
Coronavirus disease-19 (COVID-19) vaccine response data for patients with hematologic malignancy, who carry high risk for severe COVID-19 illness, are incomplete. In a study of 551 hematologic malignancy patients with leukemia, lymphoma, and multiple myeloma, anti-SARS-CoV-2 spike IgG titers and neutralizing activity were measured at 1 and 3 months from initial vaccination. Compared with healthy controls, patients with hematologic malignancy had attenuated antibody titers at 1 and 3 months. Furthermore, patients with hematologic malignancy had markedly diminished neutralizing capacity of 26.3% at 1 month and 43.6% at 3 months, despite positive seroconversion rates of 51.5% and 68.9% at the respective time points. Healthy controls had 93.2% and 100% neutralizing capacity at 1 and 3 months, respectively. Patients with leukemia, lymphoma, and multiple myeloma on observation had uniformly blunted responses. Treatment with Bruton tyrosine kinase inhibitors, venetoclax, phosphoinositide 3-kinase inhibitors, anti-CD19/CD20-directed therapies, and anti-CD38/B-cell maturation antigen-directed therapies substantially hindered responses, but single-agent immunomodulatory agents did not. Patients with hematologic malignancy have compromised COVID-19 vaccine responses at baseline that are further suppressed by active therapy, with many patients having insufficient neutralizing capacity despite positive antibody titers. Refining vaccine response parameters is critical to guiding clinical care, including the indication for booster vaccines, for this vulnerable population.See related article by Tamari et al., p. 577. This article is highlighted in the In This Issue feature, p. 549.


Role of a health psychologist in the management of functional esophageal complaints.
Upper gastrointestinal complaints are common among patients in a gastrointestinal clinic. Outside of typical gastroesophageal reflux disease symptoms that are treated with medication, the symptom presentations of esophageal patients, particularly those with functional conditions, are often difficult to treat and account for high health-care utilization. This manuscript describes the role of a health psychologist in the treatment of esophageal disorders using behavioral medicine interventions. Observations over the course of a 1-year period indicate that the sample presents with a relatively low level of psychological distress but reports negative effects of their symptoms on health-related quality of life. Five case examples of commonly treated disorders (globus, non-cardiac chest pain, functional dysphagia, rumination syndrome, supragastric belching) are described to highlight how behavioral treatment can improve patients' symptoms, decrease health-care utilization, and improve overall quality of life in a timely and relatively simple manner. Successful treatment outcomes are associated with a collaborative working alliance between patient, health psychologist, and gastroenterologist. Results indicate the benefit of referring appropriate esophageal patients to a health psychologist with specialization in gastroenterology.


How GPs implement clinical guidelines in everyday clinical practice--a qualitative interview study.
Clinical guidelines are considered to be essential for improving quality and safety of health care. However, interventions to promote implementation of guidelines have demonstrated only partial effectiveness and the reasons for this apparent failure are not yet fully understood. To investigate how GPs implement clinical guidelines in everyday clinical practice and how implementation approaches differ between practices. Individual semi-structured open-ended interviews with seven GPs who were purposefully sampled with regard to gender, age and practice form. Interviews were recorded, transcribed verbatim and then analysed using systematic text condensation. Analysis of the interviews revealed three different approaches to the implementation of guidelines in clinical practice. In some practices the GPs prioritized time and resources on collective implementation activities and organized their everyday practice to support these activities. In other practices GPs discussed guidelines collectively but left the application up to the individual GP whilst others again saw no need for discussion or collective activities depending entirely on the individual GP's decision on whether and how to manage implementation. Approaches to implementation of clinical guidelines vary substantially between practices. Supporting activities should take this into account.


Effects of curved mirror misalignment in a folded waveguide.
We have previously discussed the transmission and coupling losses of a Gaussian beam in a folded waveguide structure made of two hollow square waveguides placed symmetrically above a spherical mirror with a nonzero on-axis angle between. Here we investigate the additional loss due to the misalignment of the mirror (axial displacement and/or angular tilt). The theoretical results are in good agreement with experiment and provide alignment tolerances for folded waveguide design.


Role of particle interaction on distribution of liver lysosomes in colloidal silica.
Rat liver mitochondrial-lysosomal fractions were separated on gradients of colloidal silica. Lysosomal enzymes were distributed bimodally. The dense peak (1.117 g/ml) was nearly free from contaminants; beta-N-acetyl-D-glucosaminidase was enriched nearly 60-fold. By contrast, the buoyant peak (1.085 g/ml) co-sedimented with mitochondria, microsomes, peroxisomes, and Golgi particles. Decreasing the amount of protein layered on the gradient medium or dispersing a full sample through it shifted lysosomal marker from the buoyant to the dense peak. Thus the majority of lysosomes in the two peaks appeared to have equivalent densities. Electron microscopic examination of particles separated from gradients with layered samples showed that the dense peak contained most of the dense bodies, whereas the buoyant peak was relatively enriched in autophagic vacuoles. Dispersion, however, shifted autophagic vacuoles from the buoyant to the dense peak without affecting the distribution of dense bodies. We conclude that the bulk of buoyant particles act as a sieve to retard the density equilibration of autophagic vacuoles without specifically affecting other lysosomal enzyme-containing components.


Efficacy and Safety of Empagliflozin in the Management of Diabetes Mellitus in Heart Transplant Recipients.
Type 2 diabetes mellitus (T2DM) is prevalent in patients undergoing heart transplant, and in those without preexisting T2DM, posttransplant diabetes mellitus may develop. Both T2DM and posttransplant diabetes mellitus have been associated with increased morbidity and mortality following heart transplantation. Empagliflozin is an effective glucose-lowering therapy that reduces the incidence of major cardiovascular events in patients with T2DM. The safety and efficacy of empagliflozin in transplant patients with diabetes mellitus has yet to be established. Clinical outcomes were retrospectively examined in 22 heart transplant recipients treated with empagliflozin and compared with those of 79 heart transplant patients with diabetes mellitus receiving alternative glucose-lowering therapies. Three adverse events were recorded in empagliflozin-treated patients, leading to treatment discontinuation in 1. There were no genitourinary infections. Treatment with empagliflozin for 12 months was associated with reductions in weight, body mass index, glycated hemoglobin, and frusemide dose that were not seen in the control group. There were no large changes observed in blood pressure (systolic or diastolic) or renal function (serum urea, creatinine, or estimated glomerular filtration rate) after 12 months of treatment with empagliflozin or alternative glucose-lowering therapies. Empagliflozin appears safe and effective in the management of selected patients with diabetes mellitus following heart transplantation.


Expression of the hepatocellular chloride-dependent sulfobromophthalein uptake system in Xenopus laevis oocytes.
The expression of the basolateral chloride-activated organic anion uptake system of rat hepatocytes has been studied in Xenopus laevis oocytes. Injection of oocytes with rat liver poly(A)+RNA resulted in the functional expression of chloride-dependent sulfobromophthalein (BSP) uptake within 3-5 d. This expressed chloride-dependent BSP uptake system exhibited saturation kinetics (apparent Km approximately 6.2 microM) and efficiently extracted BSP from its binding sites on BSA. Furthermore, the chloride-activated portion of BSP uptake was inhibited by bilirubin (10 microM; inhibition 53%), 4,4'-diisothiocyano-2,2-disulfonic acid stilbene (DIDS, 100 microM; 80%), taurocholate (100 microM; 80%), and cholate (200 microM; 95%). In contrast to results with total rat liver mRNA, injection of mRNA derived from the Na+/bile acid cotransporter cDNA (Hagenbuch, B., B. Stieger, M. Foguet, H. Lübbert, and P. J. Meier. 1991. Proc. Natl. Acad. Sci. USA. In press.) had no effect on BSP uptake into oocytes. Size fractionation of total rat liver mRNA revealed that a 2.0- to 3.5-kb size-class mRNA was sufficient to express the hepatic chloride-dependent BSP uptake system. These data indicate that "expression cloning" in oocytes represents a promising approach to ultimately clone the cDNA coding for the hepatocyte high affinity, chloride-dependent organic anion uptake system. Furthermore, the results confirm that the Na+/bile acid cotransport system does not mediate BSP uptake.


Detection of amyloid beta peptides in body fluids for the diagnosis of alzheimer's disease: Where do we stand?
Alzheimer's disease (AD) is an incurable neurodegenerative disease characterized by progressive decline of cognitive abilities. Amyloid beta peptides (Aβ), Tau proteins and the phosphorylated form of the Tau protein, p-Tau, are the core pathological biomarkers of the disease, and their detection for the diagnosis of patients is progressively being implemented. However, to date, their quantification is mostly performed on cerebrospinal fluid (CSF), the collection of which requires an invasive lumbar puncture. Early diagnosis has been shown to be important for disease-modifying treatment, which is currently in development, to limit the progression of the disease. Nevertheless, the diagnosis is often delayed to the point where the disease has already progressed, and the tools currently available do not allow for a systematic follow-up of patients. Thus, the search for a molecular signature of AD in a body fluid such as blood or saliva that can be collected in a minimally invasive way offers hope. A number of methods have been developed for the quantification of core biomarkers, especially in easily accessible fluids such as the blood, that improve their accuracy, specificity and sensitivity. This review summarizes and compares these approaches, focusing in particular on their use for Aβ detection, the earliest biomarker to be modified in the course of AD. The review also discusses biomarker quantification in CSF, blood and saliva and their clinical applications.


From Toxoplasmosis to Schizophrenia via NMDA Dysfunction: Peptide Overlap between Toxoplasma gondii and N-Methyl-d-Aspartate Receptors As a Potential Mechanistic Link.
The present work aims at investigating how Toxoplasma gondii (T. gondii) infection may be linked to N-methyl-d-aspartate receptor (NMDAR) dysfunction in schizophrenia and related disorders and puts forward the hypothesis that immune responses against T. gondii may involve NMDARs. Indeed, the analysis of the protozoan proteome and NMDAR subunits for peptide commonalities shows a massive peptide overlap and supports the possibility that anti-T. gondii immune responses raised during active protozoan infection may cross-react with host NMDARs, determining disruption of neural circuits and cognitive deficits. In particular, the NMDA 2D subunit, which is mainly expressed in parvalbumin-positive interneurons, appears to be a hotspot for potential T. gondii-induced cross-reactive immune attacks.


[220.127.116.11]Paracyclophanetetraenes (PCTs): cyclic structural analogues of poly( p‑phenylene vinylene)s (PPVs).
Background: Poly( p-phenylene vinylene)s ( PPVs) and [22.214.171.124]paracyclophanetetraene ( PCT) are both composed of alternating π-conjugated para-phenylene and vinylene units. However, while the former constitute a class of π-conjugated polymers that has been used in organic electronics for decades, the latter is a macrocycle that only recently revealed its potential for applications such as organic battery electrodes. The cyclic structure endows PCT with unusual properties, and further tuning of these may be required for specific applications. Methods: In this article, we adopt an approach often used for tuning the properties of PPVs, the introduction of alkoxy (or alkylthio) substituents at the phenylene units, for tuning the optoelectronic properties of PCT. The resulting methoxy- and methylthio-substituted PCTs, obtained by Wittig cyclisation reactions, are studied by UV-vis absorption, photoluminescence, and cyclic voltammetry measurements, and investigated computationally using the visualisation of chemical shielding tensors (VIST) method. Results: The measurements show that substitution leads to slight changes in terms of absorption/emission energies and redox potentials while having a pronounced effect on the photoluminescence intensity. The computations show the effect of the substituents on the ring currents and chemical shielding and on the associated local and global (anti)aromaticity of the macrocycles, highlighting the interplay of local and global aromaticity in various electronic states. Conclusions: The study offers interesting insights into the tuneability of the properties of this versatile class of π-conjugated macrocycles.


Effects of butylated hydroxyanisole on the tumorigenicity and metabolism of N-nitrosodimethylamine and N-nitrosopyrrolidine in A/J mice.
Female A/J mice were maintained on NIH-07 diet or on NIH-07 diet containing butylated hydroxyanisole (BHA, a mixture of 2- and 3-tert-butyl-4-hydroxyanisole), 5 mg/g, for 1 week prior to and during 10 weeks of treatment with N-nitrosodimethylamine (NDMA) or N-nitrosopyrrolidine (NPYR), administered in the drinking water. Twenty weeks after nitrosamine treatment ended, mice were sacrificed and lung adenomas were counted. BHA inhibited NDMA tumorigenesis but enhanced NPYR tumorigenesis. Treatment of A/J mice for three weeks with BHA (5 mg/g) added to semisynthetic diet increased whole lung microsomal alpha-hydroxylation of NDMA and NPYR, as measured by aldehyde production, and increased lung and hepatic glutathione-S-transferase activities. No evidence was found for formation of S-methylglutathione in incubations with NDMA and hepatic supernatants obtained from BHA treated or control mice. Four h after gavage of NDMA, levels of 7-methylguanine in the lung DNA of mice treated with BHA were higher than in the lung DNA of control mice, but levels of O6-methylguanine in the two groups were the same. The results of this study indicate that BHA treatment increases the microsomal metabolic alpha hydroxylation of both NDMA and NPYR, but has differential effects on their tumorigenic activities.


Enculturing brains through patterned practices.
Recent findings in neuroscience have shown differential patterns in brain activity in response to similar stimuli and activities across cultural and social differences. This calls for a framework to understand how such differences may come to be implemented in brains and neurons. Based on strands of research in social anthropology, we argue that human practices are characterized by particular patterns, and that participating in these patterns orders how people perceive and act in particular group- and context-specific ways. This then leads to a particular patterning of neuronal processes that may be detected using e.g. brain imaging methods. We illustrate this through (a) a classical example of phoneme perception (b) recent work on performance in experimental game play. We then discuss these findings in the light of predictive models of brain function. We argue that a 'culture as patterned practices' approach obviates a rigid nature-culture distinction, avoids the problems involved in conceptualizing 'culture' as a homogenous grouping variable, and suggests that participating as a competent participant in particular practices may affect both the subjective (first person) experience and (third person) objective measures of behavior and brain activity.


Telomere length in relation to insulin resistance, inflammation and obesity among Arab youth.
The aim of this study was to determine the associations of telomere length to markers of obesity, insulin resistance and inflammation in Saudi children. A total of 69 boys and 79 girls, aged 5-12 years, participated in this cross-sectional study. Anthropometrics were measured. Serum glucose and lipid profile were measured using routine laboratory methods. Serum insulin, leptin, adiponectin, resistin, tumour necrosis factor-alpha and active plasminogen activator inhibitor 1 were quantified using customized multiplex assay kits. C-reactive protein and angiotensin II were quantified using ELISA. Leucocyte telomere length was examined by quantitative real time PCR utilizing IQ cycler. Mean telomere length was significantly shorter in obese boys compared with their lean counterparts (p = 0.049), not in girls. It was not associated to insulin resistance, adipocytokines and markers of inflammation. In girls, the significant predictor of telomere length was waist circumference, explaining 24% of variance (p = 0.041) while in boys, systolic blood pressure explained 84% of the variance (p = 0.01). Childhood obesity in boys corresponds to shorter leucocyte telomere length which is not evident in girls. The association of leucocyte telomere length to blood pressure and waist circumference in children suggests clinical implications as to the contribution of these parameters in premature ageing.


Recombination and gene flux caused by gene conversion and crossing over in inversion heterokaryotypes.
A theoretical analysis of the effects of inversions on recombination and gene flux between arrangements caused by gene conversion and crossing over was carried out. Two different mathematical models of recombination were used: the Poisson model (without interference) and the Counting model (with interference). The main results are as follows. (1) Recombination and gene flux are highly site-dependent both inside and outside the inverted regions. (2) Crossing over overwhelms gene conversion as a cause of gene flux in large inversions, while conversion becomes relatively significant in short inversions and in regions around the breakpoints. (3) Under the Counting model the recombination rate between two markers depends strongly on the position of the markers along the inverted segment. Two equally spaced markers in the central part of the inverted segment have less recombination than if they are in a more extreme position. (4) Inversions affect recombination rates in the univerted regions of the chromosome. Recombination increases in the distal segment and decreases in the proximal segment. These results provide an explanation for a number of observations reported in the literature. Because inversions are ubiquitous in the evolutionary history of many Drosophila species, the effects of inversions on recombination are expected to influence DNA variation patterns.


Resting energy expenditure and nutritional state in patients with liver cirrhosis before and after liver transplantation.
Resting energy expenditure (REE), body composition, and the biochemical parameters of liver function were measured in 26 patients before and 432 days (range: 103-1022 days) after liver transplantation (LTX). PreLTX REE was variable (mean: 1638 +/- 308 kcal/day, range: 1220-2190 kcal/day or +10 +/- 11% of Harris Benedict = HB prediction, range: -19 - +33%) and was closely related to body cell mass (r = 0.66, p < 0.0003). PostLTX REE was variable (mean: 1612 +/- 358 kcal/day, range: 1010-2490 kcal/day or +5 +/- 15% of HB prediction, range: -20 - +37%) and was closely related to body cell mass (r = 0.65, p < 0.0006). When compared with preLTX values only small changes in mean REE (-71 +/- 43 kcal/day) and a close correlation between pre and postLTX REE (r = 0.82, p < 0.001) were observed. In contrast to REE, changes in body weight were highly variable (-16.5 - +32.7 kg/year). This variance was not explained by the number of postoperative complications, pre and postLTX liver function, possible graft rejection and/or hepatitis reinfection. Pre-operative hypermetabolism (i.e. REE >+20% of HB prediction) was associated with postoperative hypermetabolism and a reduced liver function before and after LTX. Hypermetabolic patients had a poorer nutritional outcome after LTX (weight change: 0 +/- 8.4 kg/year) when compared with normometabolic controls (weight change: +5.7 +/- 7.4 kg/year; p < 0.05). There was no significant association between deviations in pre and postLTX REE and changes in body weight. When corrected for changes in the nutritional state our data provide evidence for the persistence of resting energy expenditure in liver transplant patients.


[Characteristics of the strategy of intensive care of unconscious children].
Three major groups can be distinguished among children hospitalized in resuscitation and intensive care wards in a state of unconsciousness: patients with bacterial meningitis and meningoencephalitis, viral meningoencephalitis, and noninfectious involvement of the NCS. Common strategy of treatment of these patients is characterized by some specific features: etiotropic antibiotic therapy in group 1 patients, antiviral drugs in group 2, and mainly symptomatic measures in group 3. Immunotherapy used in each of these groups is substitute in group 1, immunomodulating and immunostimulating in group 2 and even more so in group 3. Better results in group 1 are due to highly effective etiotropic antibiotics and drugs.


Job satisfaction in psychiatric nursing.
In recent years, mental health services across Europe have undergone major organizational change with a move from institutional to community care. In such a context, the impact of change on the job satisfaction of psychiatric nurses has received little attention in the literature. This paper reports on the job satisfaction of psychiatric nurses and data were collected in 2003. The population of qualified psychiatric nurses (n = 800) working in a defined geographical health board area was surveyed. Methodological triangulation with a between-methods approach was used in the study. Data were collected on job satisfaction using a questionnaire adopted from the Occupational Stress Indicator. A response rate of 346 (43%) was obtained. Focus groups were used to collect qualitative data. Factors influencing levels of job satisfaction predominantly related to the nurses work location. Other factors influencing job satisfaction included choice of work location, work routine, off duty/staff allocation arrangements, teamwork and working environment. The results of the study highlight to employers of psychiatric nurses the importance of work location, including the value of facilitating staff with choices in their working environment, which may influence the recruitment and retention of nurses in mental health services.


Effects of seasonal change in rugby league on the incidence of injury.
To assess the effects that the recent seasonal change in rugby league from winter to summer has had on the incidence of injury. All injuries that occurred during games and training were recorded throughout four consecutive seasons (three winter, one summer) for the first team squad of a British professional rugby league club. Data included nature of injury, days lost as the result of injury, player position, month, and season. An increasing incidence of injury over the four seasons was observed, with the summer seasons having an incidence almost double that of the first winter season recorded (696.8 per 1000 hours and 363.55 per 1000 hours respectively). The severity of injury was shown to decline over the four seasons. Most injuries were incurred in matches (74.9%) rather than during training (25.1%). The findings show an increasing incidence of injury in summer rugby but with decreased severity. It is not known if this is the result of playing back to back rugby seasons or is a consequence of a change in playing season and ground conditions.


Effects of hypochlorous acid and ascorbic acid on conductance, permeability, and structure of equine colonic mucosa in vitro.
To study effects of hypochlorous acid (HOCl) on equine colonic mucosa in vitro, and determine whether addition of ascorbic acid protects against the effects. 6 healthy horses and ponies. Short-circuit current was measured in mucosa mounted in Ussing chambers. Incubation conditions were: control (no additions); 5 mM HOCl; 1 mM HOCl; same and 5 mM ascorbic acid; 3 mM HOCl; 3 mM HOCl and 5 mM ascorbic acid; 7 mM HOCl; and 7 mM HOCl plus 5 mM ascorbic acid. Permeability was measured with [3H]mannitol and, at the conclusion of each experiment, tissues were examined microscopically to assess the effects of HOCl and ascorbic acid, alone or in combination. Short-circuit current and conductance increased transiently in response to 1 mM HOCl. Tissues had mild surface epithelial damage, as evident by swelling and separation of isolated cells. These changes were abolished when tissues were coincubated with 5 mM ascorbic acid and 1 mM HOCl. At 3 and 7 mM concentrations, HOCl caused marked increase in tissue conductance, short-circuit current, and permeability to mannitol; these changes were associated with histologic damage. Again, coincubation with 5 mM ascorbic acid protected against these changes. Additional studies indicated that the effects of HOCl and the protective effects of ascorbic acid were not mediated through changes in pH. HOCl in low concentrations is capable or increasing the short-circuit current in equine colon, possibly by increasing secretions; however, higher concentrations can cause tissue damage. The addition of 5 mM ascorbic acid blocks these changes. The concentration of HOCl produced by activated neutrophils could damage equine colonic mucosa and potentially contribute to or cause reperfusion injury. The ability of ascorbic acid to ameliorate this injury in an in vitro setting offers a potential method for pharmacologic evaluation of this injury and for treatment.


Prenatal diagnosis and natural history of fetuses presenting with pleural effusion.
To describe the natural history of fetuses presenting with pleural effusion. Between January 2005 and December 2009 all fetuses diagnosed with pleural effusion were followed up. Fetuses were divided into three groups: I, isolated pleural effusion; II, associated structural anomalies but normal karyotype; and III, chromosomal anomalies. Univariate and multivariate analyses were performed to determine the association between prenatal ultrasound findings and perinatal death. Fifty-six fetuses were included in the study. Associated structural or chromosomal anomalies occurred in 75.0% (42/56) of cases. Bilateral pleural effusion and fetal hydrops were associated with each other (p < 0.01) and with perinatal death (p < 0.05). Multivariate analysis indicated that only the presence of associated abnormalities was a statistical determinant of perinatal death (OR, 3.56; 95% CI, 1.48-5.64; p < 0.01). Fetal pleural effusion is often associated with other abnormalities, and this association has poor perinatal outcome.


A validated collection of mouse monoclonal antibodies to human glycosyltransferases functioning in mucin-type O-glycosylation.
Complex carbohydrates serve a wide range of biological functions in cells and tissues, and their biosynthesis involves more than 200 distinct glycosyltransferases (GTfs) in human cells. The kinetic properties, cellular expression patterns and subcellular topology of the GTfs direct the glycosylation capacity of a cell. Most GTfs are ER or Golgi resident enzymes, and their specific subcellular localization is believed to be distributed in the secretory pathway according to their sequential role in the glycosylation process, although detailed knowledge for individual enzymes is still highly fragmented. Progress in quantitative transcriptome and proteome analyses has greatly advanced our understanding of the cellular expression of this class of enzymes, but availability of appropriate antibodies for in situ monitoring of expression and subcellular topology have generally been limited. We have previously used catalytically active GTfs produced as recombinant truncated secreted proteins in insect cells for generation of mouse monoclonal antibodies (mAbs) to human enzymes primarily involved in mucin-type O-glycosylation. These mAbs can be used to probe subcellular topology of active GTfs in cells and tissues as well as their presence in body fluids. Here, we present several new mAbs to human GTfs and provide a summary of our entire collection of mAbs, available to the community. Moreover, we present validation of specificity for many of our mAbs using human cell lines with CRISPR/Cas9 or zinc finger nuclease (ZFN) knockout and knockin of relevant GTfs.


The large isoforms of A-kinase anchoring protein 18 mediate the phosphorylation of inhibitor-1 by protein kinase A and the inhibition of protein phosphatase 1 activity.
Inhibitor-1 (I-1) is phosphorylated on threonine residue 35 (Thr35) by the cAMP-dependent protein kinase (PKA), inducing the potent inhibition of the serine-threonine-specific protein phosphatase 1 (PP1). We now report that the formation of a signaling complex containing PKA and I-1 by the A-kinase anchoring protein 18 (AKAP18) facilitates this regulation in cells. AKAP18 directly bound I-1, and AKAP18/I-1 complexes were isolated from both rat heart extract and transfected heterologous cells. It is noteworthy that prevention of PKA binding to the AKAP18 scaffold decreased I-1 phosphorylation by 48% in cells. Moreover, the I-1 target PP1 was also associated with AKAP18 complexes. The cAMP-mediated inhibition of phosphatase activity was contingent on PKA binding to the scaffold. These observations reveal an additional level of complexity in PP1 regulation because of its association with AKAP18 multimolecular signaling complexes and suggest that targeting of AKAP18 complexes may be an alternative method to alter phosphatase activity and modulate specific substrate dephosphorylation.


Disparities in Perinatal and Emergency Care Receipt Among Women With Perinatal Opioid Use Disorder in Medicaid, 2007 to 2012.
This study aimed to better understand receipt of perinatal and emergency care among women with perinatal opioid use disorder (OUD) and explore variation by race/ethnicity. We used 2007-2012 Medicaid Analytic eXtract (MAX) data from all 50 states and the District of Columbia to examine 6,823,471 deliveries for women 18 to 44 years old. Logistic regressions modeled the association between (1) OUD status and receipt of perinatal and emergency care, and (2) receipt of perinatal and emergency care and race/ethnicity, conditional on OUD diagnosis and controlling for patient and county characteristics. We used robust SEs, clustered at the individual level, and included state and year fixed effects. Women with perinatal OUD were less likely to receive adequate prenatal care (adjusted odds ratio [aOR], 0.45; 95% confidence interval [CI], 0.44-0.46) and attend the postpartum visit (aOR, 0.46; 95% CI, 0.45-0.47) and more likely to seek emergency care (aOR, 1.48; 95% CI, 1.45-1.51) than women without perinatal OUD. Among women with perinatal OUD, Black, Hispanic, and American Indian and Alaskan Native (AI/AN) women were less likely to receive adequate prenatal care (aOR, 0.68 [95% CI, 0.64-0.72]; aOR, 0.86 [95% CI, 0.80-0.92]; aOR, 0.71 [95% CI, 0.64-0.79]) and attend the postpartum visit (aOR, 0.85 [95% CI, 0.80-0.91]; aOR, 0.86 [95% CI, 0.80-0.93]; aOR, 0.83 [95% CI, 0.73-0.94]) relative to non-Hispanic White women. Black and AI/AN women were also more likely to receive emergency care (aOR, 1.13 [95% CI, 1.05-1.20]; aOR, 1.12 [95% CI, 1.00-1.26]). Our findings suggest that women with perinatal OUD, in particular Black, Hispanic, and AI/AN women, may be missing opportunities for preventive care and comprehensive management of their physical and behavioral health during pregnancy.


P-selectin glycoprotein ligand-1 as a potential target for humoral immunotherapy of multiple myeloma.
Monoclonal antibodies (mAbs), successfully adopted in the treatment of several haematological malignancies, have proved almost ineffective in multiple myeloma (MM), because of the lack of an appropriate antigen for targeting and killing MM cells. Here, we demonstrate that PSGL1, the major ligand of P-Selectin, a marker of plasmacytic differentiation expressed at high levels on normal and neoplastic plasma cells, may represent a novel target for mAb-mediated MM immunotherapy. The primary effectors of mAb-induced cell-death, complement-mediated lysis (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC), were investigated using U266B1 and LP1 cell-lines as models. Along with immunological mechanisms, the induction of apoptosis by PSGL1 cross-linking was assessed. The anti-PSGL1 murine mAb KPL1 induced death of MM cells in a dose- and time-dependent fashion and mediated a significant amount of ADCC. KPL1 alone mediated C1q deposition on target cells but proved unable to induce CDC due to inhibition of the lytic activity of complement by membrane complement regulators (mCRP) expressed on the cell surface. Consistently, CDC was induced by KPL1 upon mCRP blockage. Our results suggest a role for PSGL1 in MM humoral immunotherapy and support further in vivo studies assessing the effects of anti-PSGL1 mAbs on MM growth and interaction with the bone marrow microenvironment.


Comparative analysis of anterior cervical discectomy and fusion in the inpatient versus outpatient surgical setting.
OBJECTIVEIn a large, consecutive series of patients treated with anterior cervical discectomy and fusion (ACDF) performed by a single surgeon, the authors compared the clinical and surgical outcomes of patients who underwent ACDF in an inpatient versus outpatient setting.METHODSPatients undergoing primary ACDF were retrospectively reviewed and stratified by surgical setting: hospital or ambulatory surgical center (ASC). Data regarding perioperative characteristics, including hospital length of stay and complications, were collected. Neck Disability Index (NDI) and visual analog scale (VAS) scores were used to analyze neck and arm pain in the preoperative period and at 6 weeks, 3 months, 6 months, and 12 months postoperatively. Postoperative outcomes were compared using chi-square analysis and linear regression.RESULTSThe study included 272 consecutive patients undergoing a primary ACDF, of whom 172 patients underwent surgery at a hospital and 100 patients underwent surgery at an ASC. Patients undergoing ACDF in the hospital setting were older, more likely to be diabetic, and had a higher comorbidity burden. Patients receiving treatment in the ASC were more likely to carry Workers' Compensation insurance. Patients in the hospital cohort were more likely to have multilevel procedures, had greater blood loss, and experienced a longer length of stay. In the hospital cohort, 48.3% of patients were discharged within 24 hours, while 43.0% were discharged between 24 and 48 hours after admission. Both cohorts had similar VAS pain scores on postoperative day (POD) 0; however, the hospital cohort consumed more narcotics on POD 0. One patient in the ASC cohort had a pretracheal hematoma that was evacuated immediately in the same surgical center. There were 8 cases of dysphagia in the hospital cohort and 3 cases in the ASC cohort, all of which resolved before the 6-month follow-up. Both cohorts demonstrated similar NDI and VAS neck and arm pain scores preoperatively and at every postoperative time point.CONCLUSIONSAlthough patients undergoing ACDF in the hospital setting were older, had a greater comorbidity burden, and underwent surgery on more levels than patients undergoing ACDF at an outpatient center, this study demonstrated comparable surgical and clinical outcomes for both patient groups. Based on the results of this single surgeon's experience, 1- to 2-level ACDFs may be performed successfully in the outpatient setting in appropriately selected patient populations.


Granulocyte-colony stimulating factor alleviates perinatal hypoxia-induced decreases in hippocampal synaptic efficacy and neurogenesis in the neonatal rat brain.
Using various animal models, studies have greatly expanded our understanding of perinatal hypoxia-induced neuronal injury in the newborn at the cellular/molecular levels. However, the synapse-basis pathogenesis and therapeutic strategy for such detrimental alterations in the neonatal brain remain to be addressed. We investigated whether the damaged synaptic efficacy and neurogenesis within hippocampal CA1 region (an essential integration area for mammalian learning and memory) of the neonatal rat brain after perinatal hypoxia were restored by granulocyte-colony stimulating factor (G-CSF) therapy. Ten-day-old (P10) rat pups were subjected to experimentally perinatal hypoxia. G-CSF (10, 30, or 50 μg/kg, single injection/d, P11-16) was s.c. administered to neonatal rats which were analyzed on P17. Perinatal hypoxia reduced the expression in pRaf-pERK1/2-pCREB(Ser-133) signaling, the synaptic complex of postsynaptic density protein-95 (PSD-95) with N-methyl-D-aspartate receptor (NMDAR) subunits (NR1, NR2A, and NR2B), synaptic efficacy, and neurogenesis. A representatively effective dosage of G-CSF (30 μg/kg) alleviated the perinatal hypoxia-induced detrimental changes and improved the performance in long-term cognitive function. In summary, our results suggest a novel concept that synaptic efficacy defects exist in the neonatal brain previously exposed to perinatal hypoxia and that G-CSF could be a clinical potential for the synapse-basis recovery in the perinatal hypoxia suffers.


[Changes in the immune status indices of patients with diffuse toxic goiter in different stages of the disease].
The purpose was to study some indices of immunity in the period of mercazolyl therapy and during DTG remission. T4, T3, TSH and some indices of the immunological status: RFCc, RFCl, RFCn, stimulation index (SI) in the lymphocyte blast transformation reaction, serum immunoglobulins (Ig A, M, G) were studied in 44 patients. In diffuse toxic goiter (DTG) RFCc, RFCl and RFCn were two times and more as less, the Ig G level was raised 1.5-fold. Mercazolyl therapy corrected RFCc, RFCn whereas RFCl remained lowered. The Ig G level correlated with the T4 and T3 levels. In the DTG patients an initial drop of SI (50%) was undetectable. Mercazolyl therapy raised SI by the 4-5th week of treatment (113%). SI returned to normal by the 10-12th week. In DTG remission most of the immunologic indices returned to normal.


Cell-to-cell communication in guided bone regeneration: molecular and cellular mechanisms.
This overview provides insights into the molecular and cellular mechanisms involved in guided bone regeneration, in particular focusing on aspects presented in the 3D movie, Cell-To-Cell Communication in Guided Bone Regeneration. The information presented here is based almost exclusively on genetic mouse models in which single genes can be deleted or overexpressed, even in a specific cell type. This information needs to be extrapolated to humans and related to aspects relevant to graft consolidation under the clinical parameters of guided bone regeneration. The overview follows the ground tenor of the Cell-To-Cell Communication series and focuses on aspects of cell-to-cell communication in bone regeneration and guided bone regeneration. Here, we discuss (1) the role of inflammation during bone regeneration, including (2) the importance of the fibrin matrix, and (3) the pleiotropic functions of macrophages. We highlight (4) the origin of bone-forming osteoblasts and bone-resorbing osteoclasts as well as (5) what causes a progenitor cell to mature into an effector cell. (6) We touch on the complex bone adaptation and maintenance after graft consolidation and (7) how osteocytes control this process. Finally, we speculate on (8) how barrier membranes and the augmentation material can modulate graft consolidation.


Altered Sleep Microarchitecture and Cognitive Impairment in Patients With Temporal Lobe Epilepsy.
To investigate the spatiotemporal characteristics of sleep waveforms in temporal lobe epilepsy (TLE) and examine their association with cognition. In this retrospective, cross-sectional study, we examined overnight EEG data from adult patients with TLE and nonepilepsy comparisons (NECs) admitted to the epilepsy monitoring unit at Mass General Brigham hospitals. Automated algorithms were used to characterize sleep macroarchitecture (sleep stages) and microarchitecture (spindles, slow oscillations [SOs]) on scalp EEG and to detect hippocampal interictal epileptiform discharges (hIEDs) from foramen ovale electrodes simultaneously recorded in a subset of patients with TLE. We examined the association of sleep features and hIEDs with memory and executive function from clinical neuropsychological evaluations. A total of 81 adult patients with TLE and 28 NEC adult patients were included with similar mean ages. There were no significant differences in sleep macroarchitecture between groups, including relative time spent in each sleep stage, sleep efficiency, and sleep fragmentation. By contrast, the spatiotemporal characteristics of sleep microarchitecture were altered in TLE compared with NEC and were associated with cognitive impairments. Specifically, we observed a ∼30% reduction in spindle density in patients with TLE compared with NEC, which was significantly associated with worse memory performance. Spindle-SO coupling strength was also reduced in TLE and, in contrast to spindles, was associated with diminished executive function. We found no significant association between sleep macroarchitectural and microarchitectural parameters and hIEDs. There is a fundamental alteration of sleep microarchitecture in TLE, characterized by a reduction in spindle density and spindle-SO coupling, and these changes may contribute to neurocognitive comorbidity in this disorder.


On constructing radiation hybrid maps.
Radiation hybrid (RH) mapping is a somatic cell method for obtaining ordering information of markers on a chromosome, using relatively few experiments. Given the results of a typical RH experiment, finding the true order of the markers is a challenging algorithmic problem. In this work we present several simple algorithms for ordering and mapping the markers, where the input is the genomic data obtained from RH experiments. We provide a rigorous analysis of these algorithms. In particular, we show that under the standard statistical model for RH, our algorithms are "statistically consistent." That is, given enough hybrids, the algorithms do reconstruct the true markers' order (with high probability). We also prove a simple lower bound for the number of hybrids required (by any algorithm) to correctly reconstruct the order. We have implemented these algorithms, and tested them on synthetic and real data. These simulations show that for practical input sizes (number of markers and hybrids) our algorithms produce outputs that are very close to the true ordering. The simulations also indicate that the true ordering of the markers is usually not the one which minimizes the number of obligate chromosome breaks.


Advantages and applications of CAR-expressing natural killer cells.
In contrast to donor T cells, natural killer (NK) cells are known to mediate anti-cancer effects without the risk of inducing graft-versus-host disease (GvHD). In order to improve cytotoxicity against resistant cancer cells, auspicious efforts have been made with chimeric antigen receptor (CAR) expressing T- and NK cells. These CAR-modified cells express antigen receptors against tumor-associated surface antigens, thus redirecting the effector cells and enhancing tumor-specific immunosurveillance. However, many cancer antigens are also expressed on healthy tissues, potentially leading to off tumor/on target toxicity by CAR-engineered cells. In order to control such potentially severe side effects, the insertion of suicide genes into CAR-modified effectors can provide a means for efficient depletion of these cells. While CAR-expressing T cells have entered successfully clinical trials, experience with CAR-engineered NK cells is mainly restricted to pre-clinical investigations and predominantly to NK cell lines. In this review we summarize the data on CAR expressing NK cells focusing on the possible advantage using these short-lived effector cells and discuss the necessity of suicide switches. Furthermore, we address the compliance of such modified NK cells with regulatory requirements as a new field in cellular immunotherapy.


Prevalence of CDKN2A mutations in pancreatic cancer patients: implications for genetic counseling.
Germline mutations in CDKN2A have been reported in pancreatic cancer families, but genetic counseling for pancreatic cancer risk has been limited by lack of information on CDKN2A mutation carriers outside of selected pancreatic or melanoma kindreds. Lymphocyte DNA from consecutive, unselected white non-Hispanic patients with pancreatic adenocarcinoma was used to sequence CDKN2A. Frequencies of mutations that alter the coding of p16INK4 or p14ARF were quantified overall and in subgroups. Penetrance and likelihood of carrying mutations by family history were estimated. Among 1537 cases, 9 (0.6%) carried germline mutations in CDKN2A, including three previously unreported mutations. CDKN2A mutation carriers were more likely to have a family history of pancreatic cancer (P=0.003) or melanoma (P=0.03), and a personal history of melanoma (P=0.01). Among cases who reported having a first-degree relative with pancreatic cancer or melanoma, the carrier proportions were 3.3 and 5.3%, respectively. Penetrance for mutation carriers by age 80 was calculated to be 58% for pancreatic cancer (95% confidence interval (CI) 8-86%), and 39% for melanoma (95% CI 0-80). Among cases who ever smoked cigarettes, the risk for pancreatic cancer was higher for carriers compared with non-carriers (HR 25.8, P=2.1 × 10⁻¹³), but among nonsmokers, this comparison did not reach statistical significance. Germline mutations in CDKN2A among unselected pancreatic cancer patients are uncommon, although notably penetrant, especially among smokers. Carriers of germline mutations of CDKN2A should be counseled to avoid tobacco use to decrease risk of pancreatic cancer in addition to taking measures to decrease melanoma risk.


The locust genome provides insight into swarm formation and long-distance flight.
Locusts are one of the world's most destructive agricultural pests and represent a useful model system in entomology. Here we present a draft 6.5 Gb genome sequence of Locusta migratoria, which is the largest animal genome sequenced so far. Our findings indicate that the large genome size of L. migratoria is likely to be because of transposable element proliferation combined with slow rates of loss for these elements. Methylome and transcriptome analyses reveal complex regulatory mechanisms involved in microtubule dynamic-mediated synapse plasticity during phase change. We find significant expansion of gene families associated with energy consumption and detoxification, consistent with long-distance flight capacity and phytophagy. We report hundreds of potential insecticide target genes, including cys-loop ligand-gated ion channels, G-protein-coupled receptors and lethal genes. The L. migratoria genome sequence offers new insights into the biology and sustainable management of this pest species, and will promote its wide use as a model system.