Magnitude of negative arterial-portal glucose gradient alters net hepatic glucose balance in conscious dogs.
To examine the relationship between the magnitude of the negative arterial-portal glucose gradient and net hepatic glucose uptake, two groups of 42-h fasted, conscious dogs were infused with somatostatin, to suppress endogenous insulin and glucagon secretion, and the hormones were replaced intraportally to create hyperinsulinemia (3- to 4-fold basal) and basal glucagon levels. The hepatic glucose load to the liver was doubled and different negative arterial-portal glucose gradients were established by altering the ratio between portal and peripheral vein glucose infusions. In protocol 1 (n = 6) net hepatic glucose uptake was 42.2 +/- 6.7, 35.0 +/- 3.9, and 33.3 +/- 4.4 mumol.kg-1.min-1 at arterial-portal plasma glucose gradients of -4.1 +/- 0.9, -1.8 +/- 0.4, and -0.8 +/- 0.1 mM, respectively. In protocol 2 (n = 6) net hepatic glucose uptake was 26.1 +/- 2.8 and 12.2 +/- 1.7 mumol.kg-1.min-1 at arterial-portal plasma glucose gradients of -0.9 +/- 0.2 and -0.4 +/- 0.1 mM, respectively. No changes in the hepatic insulin or glucose loads were evident within a given protocol. Although net hepatic glucose uptake was lower in protocol 2 when compared with protocol 1 (26.1 +/- 2.8 vs. 33.3 +/- 4.4 mumol.kg-1.min-1) in the presence of a similar arterial-portal plasma glucose gradient (-0.9 vs. -0.8 mM) the difference could be attributed to the hepatic glucose load being lower in protocol 2 (i.e., hepatic fractional glucose extraction was not significantly different) primarily as a result of lower hepatic blood flow. In conclusion, in the presence of fixed hepatic glucose and insulin loads, the magnitude of the negative arterial-portal glucose gradient can modify net hepatic glucose uptake in vivo.


Proteomic analysis identifies cytoskeleton-interacting proteins as major downstream targets of altered folate status in the aorta of adult rat.
Mild folate deficiency and subsequently elevated plasma level of homocysteine are associated with an increased risk for vascular diseases in adults. Conversely, high intakes of folic acid (FA) may have beneficial effects on vascular function, presumably in part through homocysteine lowering. However, these effects have not yet been translated in terms of prevention or treatment of vascular pathologies. Besides, the complex biologic perturbation induced by variations of the folate supply is still not fully deciphered. We thus carried out a proteomic analysis of the aorta of adult rats after a dietary FA depletion or supplementation. Nine month-old rats were fed a FA-depleted, FA-supplemented or control diet for 8 weeks. Total proteins from adventitia-free aortas were separated by 2DE and differentially expressed proteins were identified by MS. FA depletion or supplementation resulted in significantly changed abundance of 29 spots (p < 0.05), of which 20 proteins were identified. Bioinformatic analysis revealed that most of these proteins are involved in cytoskeleton-related processes important to cell function/maintenance, assembly/organization, and movement. Our proteomic study supports that expression of proteins essential to vascular structure and, presumably, function is modulated by high intake as well as deprivation of FA.


Feasibility of clinical electron beam formation using polymer materials produced by fused deposition modeling.
The main challenge in electron external beam radiation therapy with clinical accelerators is the absence of integrated systems to form irregular fields. The current approach to provide conformal irradiation is to use additional metallic shaping blocks, with inefficient and expensive workflows. This work presents a simple method to form therapeutic electron fields using 3D printed samples. These samples are manufactured by fused deposition modeling, which can affect crucial properties, such as material homogeneity, due to the presence of residual air-filled cavities. The applicability of this method was therefore investigated with a set of experiments and Monte Carlo simulations aimed at determining the electron depth dose distribution in polymer materials. The results show that therapeutic electron beams with energies 6-20 MeV can be effectively absorbed using these polymeric samples. The model developed in this study provides a way to assess the dose distribution in such materials and to calculate the appropriate thickness of polymer samples for therapeutic electron beam formation. It is shown that for total absorption of 6 MeV electron beams the material thickness should be at least 4 cm, while this value should be at least 8 cm for 12 MeV and 11 cm for 20 MeV, respectively. The results can be used to further develop 3D printing procedures for medical electron beam profile formation, allowing the creation of a collimator or absorber with patient-specific configuration using rapid prototyping systems, thus contributing to improve the accuracy of dose delivery in electron radiotherapy within a short manufacturing time.


Failure of infused beta-hydroxybutyrate to decrease proteolysis in man.
Ketone bodies have been suggested to have a protein-sparing effect, since infusion of Na-beta-hydroxybutyrate in man decreases plasma alanine concentrations and urinary nitrogen (N) excretion. To test this hypothesis, six normal postabsorptive volunteers were infused with Na-beta-hydroxybutyrate for 3 h. Rates of glucose, leucine carbon, and alanine appearance and disappearance from the plasma space were traced with [3-3H]glucose, L-[6,6,6-2H3]leucine, and [2,3,3,3-2H4]alanine. Rates of leucine N appearance and disappearance and the rate of transfer of leucine N to alanine were assessed with [15N]leucine. During ketone body infusion, plasma alanine decreased (P less than 0.05), whereas plasma leucine increased (P less than 0.05). Rates of alanine appearance increased (5.3 +/- 0.3 to 7.8 +/- 0.6 mumol/kg X min), but the increase in its rate of disappearance was slightly greater, accounting for the decrease in plasma alanine concentration. Leucine N flux and the rate and percent of leucine N transferred to alanine increased, whereas leucine carbon flux was unchanged. To determine the effect of the alkalemia induced by Na-beta-hydroxybutyrate, four additional subjects were infused with NaHCO3. Alkalemia had no effect on leucine N or carbon flux or on the rate of appearance of alanine, but increased the rate of alanine disappearance, resulting in a decrease in the plasma alanine concentration. Since the rate of appearance of leucine carbon was unaltered during the infusion of Na-beta-hydroxybutyrate, it is unlikely that hyperketonemia per se decreases proteolysis in postabsorptive man.


[Antibiotic therapy of purulent-inflammatory diseases].
The analysis of 161 case record of the patients with purulent-inflammatory diseases was carried out. The dependence of duration of treatment on the age of a patient, character of pathology, body temperature, leukocytic count, presence of pathologic changes in the urine was established. The expediency to limit antibiotic therapy in purulent-inflammatory diseases is substantiated.


Flavor Compounds in Pixian Broad-Bean Paste: Non-Volatile Organic Acids and Amino Acids.
Non-volatile organic acids and amino acids are important flavor compounds in Pixian broad-bean paste, which is a traditional Chinese seasoning product. In this study, non-volatile organic acids, formed in the broad-bean paste due to the metabolism of large molecular compounds, are qualitatively and quantitatively determined by high-performance liquid chromatography (HPLC). Amino acids, mainly produced by hydrolysis of soybean proteins, were determined by the amino acid automatic analyzer. Results indicated that seven common organic acids and eighteen common amino acids were found in six Pixian broad-bean paste samples. The content of citric acid was found to be the highest in each sample, between 4.1 mg/g to 6.3 mg/g, and malic acid were between 2.1 mg/g to 3.6 mg/g ranked as the second. Moreover, fumaric acid was first detected in fermented bean pastes albeit with a low content. For amino acids, savory with lower sour taste including glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp) and asparagines (Asn) were the most abundant, noted to be 6.5 mg/g, 4.0 mg/g, 6.4 mg/g, 4.9 mg/g, 6.2 mg/g and 10.2 mg/g, and bitter taste amino acids followed. More importantly, as important flavor materials in Pixian broad-bean paste, these two groups of substances are expected to be used to evaluate and represent the flavor quality of Pixian broad-bean paste. Moreover, the results revealed that citric acid, glutamic acid, methionine and proline were the most important flavor compounds. These findings are agreat contribution for evaluating the quality and further assessment of Pixian broad-bean paste.


Interaction of Chrysin and Its Main Conjugated Metabolites Chrysin-7-Sulfate and Chrysin-7-Glucuronide with Serum Albumin.
Chrysin (5,7-dihydroxyflavone) is a flavonoid aglycone, which is found in nature and in several dietary supplements. During the biotransformation of chrysin, its conjugated metabolites chrysin-7-sulfate (C7S) and chrysin-7-glucuronide (C7G) are formed. Despite the fact that these conjugates appear in the circulation at much higher concentrations than chrysin, their interactions with serum albumin have not been reported. In this study, the complex formation of chrysin, C7S, and C7G with human (HSA) and bovine (BSA) serum albumins was investigated employing fluorescence spectroscopic, ultrafiltration, and modeling studies. Our major observations/conclusions are as follows: (1) Compared to chrysin, C7S binds with a threefold higher affinity to HSA, while C7G binds with a threefold lower affinity; (2) the albumin-binding of chrysin, C7S, and C7G did not show any large species differences regarding HSA and BSA; (3) tested flavonoids likely occupy Sudlow's Site I in HSA; (4) C7S causes significant displacement of Sudlow's Site I ligands, exerting an even stronger displacing ability than the parent compound chrysin. Considering the above-listed observations, the high intake of chrysin (e.g., through the consumption of dietary supplements with high chrysin contents) may interfere with the albumin-binding of several drugs, mainly due to the strong interaction of C7S with HSA.


[Articulation disorder as the initial manifestation of facioscapulohumeral muscular dystrophy in childhood].
A five year old boy who had received logopedic treatment for more than two years was seen as an outpatient because of speech retardation. He presented with myopathic face, incomplete closure of both lids and severe weakness of facial muscles, bilateral winging of scapulae and hyperlordosis. Extraocular and pharyngeal muscles were not affected. Motor and sensory nerve fibre conductions and electromyography were within normal limits. CPK was moderately elevated (320 U/I). Muscle biopsy of right deltoid muscle revealed unspecific myopathic changes. The patients brother aged 7 also presented with facial weakness, elevated CPK and neurogenic changes in EMG of deltoid muscle. Both parents were clinically and electrophysiologically unremarkable. Although problems to speak distinctly are usually not the first manifestation, we found in this family facio-scapulo-humeral muscular dystrophy.


Plant diversity changes during the postglacial in East Asia: insights from Forest Refugia on Halla Volcano, Jeju Island.
Understanding how past climate changes affected biodiversity is a key issue in contemporary ecology and conservation biology. These diversity changes are, however, difficult to reconstruct from paleoecological sources alone, because macrofossil and pollen records do not provide complete information about species assemblages. Ecologists therefore use information from modern analogues of past communities in order to get a better understanding of past diversity changes. Here we compare plant diversity, species traits and environment between late-glacial Abies, early-Holocene Quercus, and mid-Holocene warm-temperate Carpinus forest refugia on Jeju Island, Korea in order to provide insights into postglacial changes associated with their replacement. Based on detailed study of relict communities, we propose that the late-glacial open-canopy conifer forests in southern part of Korean Peninsula were rich in vascular plants, in particular of heliophilous herbs, whose dramatic decline was caused by the early Holocene invasion of dwarf bamboo into the understory of Quercus forests, followed by mid-Holocene expansion of strongly shading trees such as maple and hornbeam. This diversity loss was partly compensated in the Carpinus forests by an increase in shade-tolerant evergreen trees, shrubs and lianas. However, the pool of these species is much smaller than that of light-demanding herbs, and hence the total species richness is lower, both locally and in the whole area of the Carpinus and Quercus forests. The strongly shading tree species dominating in the hornbeam forests have higher leaf tissue N and P concentrations and smaller leaf dry matter content, which enhances litter decomposition and nutrient cycling and in turn favored the selection of highly competitive species in the shrub layer. This further reduced available light and caused almost complete disappearance of understory herbs, including dwarf bamboo.


Computed tomography imaging superimposition protocols to assess outcomes in orthognathic surgery: a systematic review with comprehensive recommendations.
A systematic review was performed to analyze the current evidence on three-dimensional (3D) computed tomography (CT) superimposition protocols used to assess dentomaxillofacial changes after orthognathic and orthofacial surgery. Accuracy, reproducibility, and efficiency were evaluated. The search was divided into Main Search (PubMed, EMBASE, Cochrane Library, LILACS, and SciELO), Grey Literature search (Google Scholar and Open Grey), and Manual search. Thirteen studies were included. Of these, 10 reported data on accuracy, 10 on reproducibility and five on efficiency. Seven proposed or evaluated methods of voxel-based superimposition, three focused on the surface-based technique, one compared surface- and voxel-based superimposition protocols, one used the maximum mutual information algorithm, and one described a landmark-based superimposition method. Cone-beam computed tomography (CBCT) was the most common imaging technique, being used in 10 studies. The accuracy of most methods was high, showing mean differences smaller than voxels' dimensions, ranging between 0.05 and 1.76 mm for translational accuracy, and 0.10-1.09° for rotational accuracy. The overall reproducibility was considered good as demonstrated by the small mean error (range: 0.01-0.26 mm) and high correlation coefficients (range: 0.53-1.00). Timing to complete virtual superimposition techniques ranged between a few seconds up to 40 min. Voxel-based superimposition protocols presented the highest accuracy and reproducibility. Moreover, superimposition protocols that used automated processes and involved only one software were the most efficient.


The role of social cognition in early trust.
Language serves as a means of learning from other people. However, communication systems allow the truth to be disguised, whether out of ignorance, fear, deviousness or mistaken judgment. Recent research has explored the extent to which young children are selective in whom they trust for information and the mechanisms that support such selectivity.


The impact of amyloid-beta and tau on prospective cognitive decline in older individuals.
Amyloid-beta (Aβ) and tau pathologies are commonly observed among clinically normal older individuals at postmortem and can now be detected with in vivo neuroimaging. The association and interaction of these proteinopathies with prospective cognitive decline in normal aging and preclinical Alzheimer's disease (AD) remains to be fully elucidated. One hundred thirty-seven older individuals (age = 76.3 ± 6.22 years) participating in the Harvard Aging Brain Study underwent Aβ (11 C-Pittsburgh compound B) and tau (18 F-flortaucipir) positron emission tomography (PET) with prospective neuropsychological assessments following PET imaging (mean number of cognitive visits = 2.8 ± 1.1). Tau and Aβ PET measures were assessed in regions of interest (ROIs) as well as vertex-wise map analyses. Cognitive change was evaluated with Memory and Executive Function composites. Higher levels of Aβ and tau were both associated with greater memory decline, but not with change in executive function. Higher cortical Aβ was associated with higher tau levels in all ROIs, independent of age, and very elevated levels of tau were observed primarily in clinically normal with elevated Aβ. A significant interaction between tau and Aβ was observed in both ROI and map-level analyses, such that rapid prospective memory decline was observed in participants who had high levels of both pathologies. Our results are consistent with the supposition that both Aβ and tau are necessary for memory decline in the preclinical stages of AD. These findings may be relevant for disambiguating aging and early cognitive manifestations of AD, and to inform secondary prevention trials in preclinical AD. Ann Neurol 2019;00:1-3 ANN NEUROL 2019;85:181-193.


GOLPH3 and YB-1 Are Novel Markers Correlating With Poor Prognosis in Prostate Cancer.
Prostate cancer is a common and aggressive cancer among men. Despite advances in the treatment, the mechanisms involved in progression are still unclear. New prognostic markers should be explored for better design of patient-specific therapeutic regimens. This study was performed on 120 patients stratified as 76 with prostatic carcinoma, 12 with low-grade prostate intraepithelial lesion, 12 with high-grade prostate intraepithelial lesion and 20 with benign prostate hyperplasia. Immunohistochemical study was done for Golgi phosphoprotein 3 (GOLPH3) and Y-box binding protein-1 (YB-1) analysis. Correlation with clinicopathological data and overall survival was analyzed. Both GOLPH3 and YB-1 showed increased expression from benign to malignant tumors. In prostatic carcinoma, cytoplasmic GOLPH3 was associated with Gleason score, stage and androgen receptor (P = 0.034, P < 0.001, and P = 0.008 respectively). Nuclear YB-1 expression was associated with Gleason score and androgen receptor (P = 0.018 and P = 0.024 respectively). Cytoplasmic YB-1 expression was associated with Gleason score, stage and androgen receptor (P = 0.008, P = 0.027, and P < 0.001 respectively). High Gleason score (P = 0.004), high stage (P < 0.001) and androgen receptor (P = 0.006) were the only detected adverse prognostic clinicopathological factors. Moderate/intense GOLPH3 and high nuclear and cytoplasmic YB-1 expression were correlated with shorter overall survival (P < 0.001, P = 0.020, and P < 0.001 respectively). In the multivariate analysis, moderate/intense GOLPH3 expression was the only predictor of overall survival (P = 0.025). High GOLPH3 and nuclear/cytoplasmic YB-1 expression correlated with poor prognosis in prostate cancer. Both markers can be promising targets for new treatment strategies.


Hepatocarcinoma: considerations on surgical treatment in a personal series of 23 patients.
Surgical treatment of primary liver tumors has undergone significant changes in recent years because of improved surgical and anesthesiological techniques and better pre- and post-operative care. We review our personal series from 1987-1995. Of 31 cases of hepatocellular carcinoma (HCC) observed in the years 1987-1995, 23 underwent curative resective surgery for a total of 24 liver resections: 6 hepatectomies; 10 segmentectomies; 4 atypical subsegmentectomies; 2 extended resections, with excision of neoplastic thrombi within the portal vein; 1 orthotopic liver transplantation in another institution, and 1 limited segmental resection for tumor recurrence. In 7 recent cases, pre-operative transcatheter arterial chemoembolization (TAE) was used. The mean survival of the 13 patients that are known to be deceased is 27 months (range: 7-114 months). Perioperative mortality was nil. Actuarial 5-year survival rate is 27%. Pre-operative TAE was used in 7 patients: 4 out of 7 lesions were significantly reduced at computed tomography (CT) scan control 21 days following TAE, while in 3 the tumor size was unchanged. Liver surgery, even major resections, has become safe with no perioperative mortality in our series. In our experience, pre-operative TAE has often produced significant reduction of the mass, but its real efficacy is still the subject of debate. TAE and percutaneous ethanol injection (PET) should be evaluated as part of combined multimodality treatment in the therapy of large lesions previously considered inoperable.


Identification of protein receptors for coronaviruses by mass spectrometry.
As obligate intracellular parasites, viruses need to cross the plasma membrane and deliver their genome inside the cell. This step is initiated by the recognition of receptors present on the host cell surface. Receptors can be major determinants of tropism, host range, and pathogenesis. Identifying virus receptors can give clues to these aspects and can lead to the design of intervention strategies. Interfering with receptor recognition is an attractive antiviral therapy, since it occurs before the viral genome has reached the relative safe haven within the cell. This chapter describes the use of an immunoprecipitation approach with Fc-tagged viral spike proteins followed by mass spectrometry to identify and characterize the receptor for the Middle East respiratory syndrome coronavirus. This technique can be adapted to identify other viral receptors.


Advances in Achieving Opioid Analgesia Without Side Effects.
Opioids are the most effective drugs for the treatment of severe pain, but they also cause addiction and overdose deaths, which have led to a worldwide opioid crisis. Therefore, the development of safer opioids is urgently needed. In this article, we provide a critical overview of emerging opioid-based strategies aimed at effective pain relief and improved side effect profiles. These approaches comprise biased agonism, the targeting of (i) opioid receptors in peripheral inflamed tissue (by reducing agonist access to the brain, the use of nanocarriers, or low pH-sensitive agonists); (ii) heteromers or multiple receptors (by monovalent, bivalent, and multifunctional ligands); (iii) receptor splice variants; and (iv) endogenous opioid peptides (by preventing their degradation or enhancing their production by gene transfer). Substantial advancements are underscored by pharmaceutical development of new opioids such as peripheral κ-receptor agonists, and by treatments augmenting the action of endogenous opioids, which have entered clinical trials. Additionally, there are several promising novel opioids comprehensively examined in preclinical studies, but also strategies such as biased agonism, which might require careful rethinking.


Legally Recognizing Reproductive Coercion while Questioning Sexual Violence Exceptionalism.
While sexual violence should not be the prerequisite for legal abortion, expanding definitions of abuse to include reproductive coercion can open avenues of access to abortion following the Dobbs decision. Understanding the increased danger and compounding challenges of intimate partner violence can inform legislative initiatives, healthcare responses, and movements for reproductive justice.


Productive herpesvirus lytic replication in primary effusion lymphoma cells requires S-phase entry.
Gammaherpesviruses establish lifelong latent infection in B lymphocytes and are the causative agent of several B-cell malignancies and lymphoproliferative disorders. While a quiescent latent infection allows these pathogens to evade immune detection, initiation of an alternative lifecycle stage, known as lytic replication, is an essential step in the production and dissemination of infectious progeny. Although cessation of cellular proliferation is an eventual consequence of lytic induction, exactly how gammaherpesviruses manipulate the cell cycle prior to amplification of viral DNA remains under debate. Here we show that the onset of Kaposi's sarcoma-associated herpesvirus (KSHV) lytic reactivation in B cells leads to S-phase accumulation and that exit from G1 is required for efficient viral DNA replication. We also show that lytic replication leads to an S-phase-specific activation of the DNA damage response (DDR) that is abrogated when lytic replication is restricted to G0/G1. Finally, we observe that expression of early lytic viral genes results in cellular replication stress with increased stalling of DNA replication forks. Overall, we demonstrate that S-phase entry is important for optimal KSHV replication, that G1 arresting compounds are effective inhibitors of viral propagation, and that lytic-induced cell-cycle arrest could occur through the obstruction of cellular replication forks and subsequent activation of the DDR.


Thermodynamic state-dependent structure-based coarse-graining of confined water.
We develop thermodynamic state-dependent single-site isotropic coarse-grained potentials to predict the structure of water confined inside graphene slit-like channels by two multiscale simulation approaches: the coarse-grained molecular dynamics (CG-MD) and the empirical potential-based quasi-continuum theory (EQT). The structurally-consistent coarse-grained potentials for the CG-MD and EQT are systematically determined from the reference all-atom SPC/E water MD (AA-MD) results. For optimization of the CG-MD potentials, the relative entropy based coarse-graining method is used, and for determination of the EQT potentials, we develop a potential of mean force matching scheme. The optimized coarse-grained potentials are found to be dependent on the thermodynamic state. They are evaluated for their ability to predict the density profile of confined water, and it is found that the results obtained by the CG-MD and EQT simulations are in good agreement with the reference AA-MD results.


Systems medicine dissection of chr1q-amp reveals a novel PBX1-FOXM1 axis for targeted therapy in multiple myeloma.
Understanding the biological and clinical impact of copy number aberrations (CNAs) on the development of precision therapies in cancer remains an unmet challenge. Genetic amplification of chromosome 1q (chr1q-amp) is a major CNA conferring an adverse prognosis in several types of cancer, including in the blood cancer multiple myeloma (MM). Although several genes across chromosome 1 (chr1q) portend high-risk MM disease, the underpinning molecular etiology remains elusive. Here, with reference to the 3-dimensional (3D) chromatin structure, we integrate multi-omics data sets from patients with MM with genetic variables to obtain an associated clinical risk map across chr1q and to identify 103 adverse prognosis genes in chr1q-amp MM. Prominent among these genes, the transcription factor PBX1 is ectopically expressed by genetic amplification and epigenetic activation of its own preserved 3D regulatory domain. By binding to reprogrammed superenhancers, PBX1 directly regulates critical oncogenic pathways and a FOXM1-dependent transcriptional program. Together, PBX1 and FOXM1 activate a proliferative gene signature that predicts adverse prognosis across multiple types of cancer. Notably, pharmacological disruption of the PBX1-FOXM1 axis with existing agents (thiostrepton) and a novel PBX1 small molecule inhibitor (T417) is selectively toxic against chr1q-amp myeloma and solid tumor cells. Overall, our systems medicine approach successfully identifies CNA-driven oncogenic circuitries, links them to clinical phenotypes, and proposes novel CNA-targeted therapy strategies in MM and other types of cancer.


Solution structure of the CD3epsilondelta ectodomain and comparison with CD3epsilongamma as a basis for modeling T cell receptor topology and signaling.
Invariant CD3 subunit dimers (CD3epsilongamma, CD3epsilondelta, and CD3zetazeta) are the signaling components of the alphabeta T cell receptor (TCR). The recently solved structure of murine CD3epsilongamma revealed a unique side-to-side interface and central beta-sheets conjoined between the two C2-set Ig-like ectodomains, with the pairing of the parallel G strands implying a potential concerted piston-type movement for signal transduction. Although CD3gamma and CD3delta each dimerize with CD3epsilon, there are differential CD3 subunit requirements for receptor assembly and signaling among T lineage subpopulations, presumably mandated by structural differences. Here we present the solution structure of the heterodimeric CD3epsilondelta complex. Whereas the CD3epsilon subunit conformation is virtually identical to that in CD3epsilongamma, the CD3delta ectodomain adopts a C1-set Ig fold, with a narrower GFC front face beta-sheet that is more parallel to the ABED back face than those beta-sheets in CD3epsilon and CD3gamma. The dimer interface between CD3delta and CD3epsilon is highly conserved among species and of similar character to that in CD3epsilongamma. Glycosylation sites in CD3delta are arranged such that the glycans may point away from the membrane, consistent with a model of TCR assembly that allows the CD3delta chain to be in close contact with the TCR alpha-chain. This and many other structural and biological features provide a basis for modeling putative TCR/CD3 extracellular domain associations. The fact that the two clusters of transmembrane helices, namely, the three CD3epsilon-CD3gamma-TCRbeta segments and the five CD3epsilon-CD3delta-TCRalpha-CD3zeta-CD3zeta segments, are presumably centered beneath the G strand-paired CD3 heterodimers has important implications for TCR signaling.


Iml3p, a component of the Ctf19 complex of the budding yeast kinetochore is required to maintain kinetochore integrity under conditions of spindle stress.
The Ctf19 multi-protein complex of the central kinetochore in Saccharomyces cerevisiae is required for precise chromosome segregation during mitosis. Of 11 proteins of this complex, at least six are required for cell survival when microtubules are defective. To find individual roles of these proteins in kinetochore stability, double deletion mutants of the corresponding genes were constructed in several combinations. The growth phenotype of all the mutants was in accordance with the current model of hierarchical assembly of kinetochore proteins except one that lacked CHL4 and IML3 genes in a tubulin-defective background (tub1-1 chl4 iml3). tub1-1 chl4 iml3 showed synergistic growth defect, decrease in minichromosome stability compared with its single mutants, and a greater accumulation of cells at the G2/M checkpoint of the cell cycle. Furthermore, in the absence of Iml3p, the two-hybrid interaction between Ctf19p (a member of the Ctf19 complex) and Dam1p (a member of the outer kinetochore DASH complex) was disrupted and the localization of Dam1p at the kinetochore was also compromised. These results indicate a role for Iml3p distinct from Chl4p at the kinetochore. Iml3p may be acting as a link between the central and the outer complexes thus contributing to a functional kinetochore.


Increased AQP2 targeting in primary cultured IMCD cells in response to angiotensin II through AT1 receptor.
Vasopressin and angiotensin II (ANG II) play a major role in renal water and Na(+) reabsorption. We previously demonstrated that ANG II AT(1) receptor blockade decreases dDAVP-induced water reabsorption and AQP2 levels in rats, suggesting cross talk between these two peptide hormones (Am J Physiol Renal Physiol 288: F673-F684, 2005). To directly address this issue, primary cultured inner medullary collecting duct (IMCD) cells from male Sprague-Dawley rats were treated for 15 min with 1) vehicle, 2) ANG II, 3) ANG II + the AT(1) receptor blocker candesartan, 4) dDAVP, 5) ANG II + dDAVP, or 6) ANG II + dDAVP + candesartan. Immunofluorescence microscopy revealed that 10(-8) M ANG II or 10(-11) M dDAVP (protocol 1) was associated with increased AQP2 labeling of the plasma membrane and decreased cytoplasmic labeling, respectively. cAMP levels increased significantly in response to 10(-8) M ANG II and were potentiated by cotreatment with 10(-11) M dDAVP. Consistent with this finding, immunoblotting revealed that this cotreatment significantly increased expression of phosphorylated AQP2. ANG II-induced AQP2 targeting was blocked by 10(-5) M candesartan. In protocol 2, treatment with a lower concentration of dDAVP (10(-12) M) or ANG II (10(-9) M) did not change subcellular AQP2 distribution, whereas 10(-12) M dDAVP + 10(-9) M ANG II enhanced AQP2 targeting. This effect was inhibited by cotreatment with 10(-5) M candesartan. ANG II-induced cAMP accumulation and AQP2 targeting were inhibited by inhibition of PKC activity. In conclusion, ANG II plays a role in the regulation of AQP2 targeting to the plasma membrane in IMCD cells through AT(1) receptor activation and potentiates the effect of dDAVP on AQP2 plasma membrane targeting.


Influence of patient, physician, and hospital factors on 30-day readmission following pancreatoduodenectomy in the United States.
IMPORTANCE It is not known whether hospital and surgeon volumes have an association with readmission among patients undergoing pancreatoduodenectomy. To evaluate patient-, surgeon-, and hospital-level factors associated with readmission. Retrospective cohort study using the Surveillance, Epidemiology, and End Results (SEER)-Medicare data with cases diagnosed from January 1, 1998, to December 31, 2005, and followed up until December 2007. Population-based cancer registry data were linked to Medicare data for the corresponding patients. A total of 1488 unique individuals who underwent a pancreatoduodenectomy were identified. Undergoing pancreatoduodenectomy at hospitals classified by volume of pancreatoduodenectomy procedures performed at the facility were either very-low, low, medium, or high volume. Undergoing pancreatoduodenectomy by surgeons classified by volume of pancreatoduodenectomy procedures performed by the surgeon were either very-low, low, medium, or high volume. In-hospital morbidity, mortality, and 30-day readmission were examined. The median age was 74 years, and 1436 patients (96.5%) had a least 1 medical comorbidity. Patients were treated by 575 distinct surgeons at 298 distinct hospitals. Length of stay was longest (median, 17 days) and 90-day mortality highest (17.2%) at very-low-volume hospitals (P < .001). Among all pancreatoduodenectomy patients, 292 (21.3%) were readmitted within 30 days of discharge. There was no effect of surgeon volume and a modest effect of hospital volume (odds ratio for highest- vs lowest-volume quartiles, 1.85; 95% CI, 1.22-2.80; P = .02). The presence of significant preoperative medical comorbidities was associated with an increased risk for hospital readmission after pancreatoduodenectomy. A comorbidity score greater than 13 had a pronounced effect on the chance of readmission following pancreatoduodenectomy (odds ratio, 2.06; 95% CI, 1.56-2.71; P < .001). The source of variation in readmission was primarily attributable to patient-related factors (95.4%), while hospital factors accounted for 4.3% of the variability and physician factors for only 0.3%. Nearly 1 in 5 patients are readmitted following pancreatoduodenectomy. While variation in readmission is, in part, attributable to differences among hospitals, the largest share of variation was found at the patient level.


Studies of the Bradyrhizobium japonicum nodD1 promoter: a repeated structure for the nod box.
Induction of nod genes in Rhizobium and Bradyrhizobium species is dependent on the presence of plant-produced flavonoids, the NodD protein, and the cis-acting nod box promoter sequence. Although the nodD (nodD1) gene in Rhizobium species is constitutively expressed, nodD1 expression in Bradyrhizobium japonicum is inducible by isoflavones in a manner similar to that of the nodYABC operon. A consensus nod box sequence is found 5' of the nodYABC operon, whereas a presumptive, nod box-like sequence is found 5' of the nodD1 gene. As an initial step toward examining the nodD1 promoter, the transcriptional start sites of the nodD1 and nodYABC operons were determined and found to be 44 and 28 bp, respectively, downstream of their respective nod box sequences. A series of deletions of the nodD1 promoter were constructed and fused to the lacZ gene. Analysis of the activity of these deletions clearly showed that the divergent nod box sequence was essential for nodD1 induction by isoflavones or soybean seed extract. The induction of nodD1 expression requires NodD1, as tested in B. japonicum and in a heterologous system, Agrobacterium tumefaciens. On the basis of these data, we analyzed the published nod box sequences and propose a new consensus sequence composed of paired 9-bp repeats. Analysis of the nodD1 nod box and synthetic constructs of the nocYABC nod box indicate that at least two 9-bp repeats are required for NodD1-mediated induction. Furthermore, insertions between the paired repeats of the nodYABC nod box suggest that orientation of the repeats on opposite faces of the DNA helix is essential for maximum nod gene expression.


Effect of m-3m3FBS on Ca2+ handling and viability in OC2 human oral cancer cells.
The effect of 2,4,6-trimethyl-N-(meta-3-trifluoromethyl-phenyl)-benzenesulfonamide (m-3M3FBS), a presumed phospholipase C activator, on cytosolic free Ca2+ concentrations ([Ca2+]i) in OC2 human oral cancer cells is unclear. This study explored whether m-3M3FBS changed basal [Ca2+]i levels in suspended OC2 cells by using fura-2 as a Ca2+-sensitive fluorescent dye. M-3M3FBS at concentrations between 10-60 μM increased [Ca2+]i in a concentration-dependent manner. The Ca2+ signal was reduced partly by removing extracellular Ca2+. M-3M3FBS-induced Ca2+ influx was inhibited by the store-operated Ca2+ channel blockers nifedipine, econazole and SK&F96365, and by the phospholipase A2 inhibitor aristolochic acid. In Ca2+-free medium, 30 μM m-3M3FBS pretreatment inhibited the [Ca2+]i rise induced by the endoplasmic reticulum Ca2+ pump inhibitors thapsigargin and 2,5-di-tert-butylhydroquinone (BHQ). Conversely, pretreatment with thapsigargin, BHQ or cyclopiazonic acid partly reduced m-3M3FBS-induced [Ca2+]i rise. Inhibition of inositol 1,4,5-trisphosphate formation with U73122 did not alter m-3M3FBS-induced [Ca2+]i rise. At concentrations between 5 and 100 μM m-3M3FBS killed cells in a concentration-dependent manner. The cytotoxic effect of m-3M3FBS was not reversed by prechelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Propidium iodide staining data suggest that m-3M3FBS (20 or 50 μM) induced apoptosis in a Ca2+-independent manner. Collectively, in OC2 cells, m-3M3FBS induced [Ca2+]i rise by causing inositol 1,4,5-trisphosphate-independent Ca2+ release from the endoplasmic reticulum and Ca2+ influx via phospholipase A2-sensitive store-operated Ca2+ channels. M-3M3FBS also induced Ca2+-independent cell death and apoptosis.


Evaluation of subclinical cardiac damage in biopsy-proven nonalcoholic fatty liver disease: a systematic review and meta-analysis.
Data on the association of nonalcoholic fatty liver disease (NAFLD) with subclinical cardiac damage are scanty. We performed a systematic review to provide comprehensive information on subclinical cardiac alterations among NAFLD subjects. PubMed and the Cochrane Library were searched to identify studies comparing subclinical cardiac damage between NAFLD and healthy adults. We also searched PROSPERO to check for any similar meta-analysis in progress in order to avoid duplication with our study. Conference abstracts and the reference lists of relevant studies and systematic reviews were perused. The Newcastle-Ottawa quality assessment scale for case-control and cohort studies were used to assess study quality. Seven studies were finally included in the meta-analysis (1 cross sectional and 6 case-control), with a total of 602 individuals (362 patients with NAFLD). Epicardial fat thickness were statistically significantly higher in patients with NAFLD than in controls (mean difference [MD] 1.17, 95% confidence interval [CI] 0.45-1.89, I 2=89%). Global longitudinal strain was lower in NAFLD, to a statistically significant degree (MD -3.17, 95%CI -5.09 to -1.24, I 2=89%). However, significant heterogeneity of the findings was observed. Our findings indicate that NAFLD is related to subclinical cardiac damage. Further studies with a larger number of biopsy-proven NAFLD patients are needed to confirm this finding. Preventive and therapeutic interventions early in the course of the disease might decrease morbidity in this high-risk patient group.


Laparoscopic-assisted spleen-preserving and pylorus-preserving total pancreatectomy for main duct type intraductal papillary mucinous tumors of the pancreas: a case report.
Minimally invasive and function-preserving pancreatectomy would be the ideal approach for benign and borderline malignant tumors of the pancreas. Total pancreatectomy can be indicated for the main duct type of intraductal papillary mucin-producing tumor (IPMT) to achieve radical resection. Recently, several studies advocating total pancreatectomy in IPMT have been published, but they are all believed to be done by conventional laparotomy. Herein, we report a case of a 72-year-old female patient who successfully underwent laparoscopic-assisted total pancreatectomy with the spleen and pylorus preserved in borderline malignant main duct type IPMT. A marginal ulcer around the duodenojejunostomy was developed, but managed by a proton-pump inhibitor. She was discharged 20 days after surgery. She was followed for more than 2 years without evidence of tumor recurrence. Her blood sugar level was well controlled by insulin pump therapy and image study showed well-preserved spleen function.


Introns encode dsRNAs undetected by RIG-I/MDA5/interferons and sensed via RNase L.
Double-stranded RNA (dsRNA), a hallmark viral material that activates antiviral interferon (IFN) responses, can appear in human cells also in the absence of viruses. We identify phosphorothioate DNAs (PS DNAs) as triggers of such endogenous dsRNA (endo-dsRNA). PS DNAs inhibit decay of nuclear RNAs and induce endo-dsRNA via accumulation of high levels of intronic and intergenic inverted retroelements (IIIR). IIIRs activate endo-dsRNA responses distinct from antiviral defense programs. IIIRs do not turn on transcriptional RIG-I/MDA5/IFN signaling, but they trigger the dsRNA-sensing pathways of OAS3/RNase L and PKR. Thus, nuclear RNA decay and nuclear-cytosolic RNA sorting actively protect from these innate immune responses to self. Our data suggest that the OAS3/RNase L and PKR arms of innate immunity diverge from antiviral IFN responses and monitor nuclear RNA decay by sensing cytosolic escape of IIIRs. OAS3 provides a receptor for IIIRs, whereas RNase L cleaves IIIR-carrying introns and intergenic RNAs.


Expression of the neurosecretory process in PC12 cells is governed by REST.
The neurosecretory process is acquired during differentiation and can be lost en block by differentiated cells. To investigate the role of REST/NRSF, a transcription repressor, in the maintenance of the process we studied two PC12 clones, one wt and one defective, expressing low and high levels of endogenous RE-1 silencing transcription (factor) (REST), respectively. Stable transfection of constructs demonstrated that REST represses 10 genes coding for proteins of neurosecretory vesicles and their exocytosis, eight including and two lacking the REST-binding sequence, RE-1. Of these genes, those of chromogranins were strongly repressed by fewfold increases of REST, those of VAMP2 and syntaxin1a required much higher levels. Moreover, in wt cells transfected with an active construct the dense-core vesicles, still competent for regulated exocytosis, were much smaller, with lighter cores; in defective cells, the dominant-negative construct induced the rescue of many vesicle/exocytosis genes but not of those of chromogranins. Small dense-core vesicles, exocytized upon stimulation, were rescued when the construct-transfected defective cells were transfected also with chromograninA or treated with trichostatinA, a blocker of histone deacetylases. Our results identify REST, working by direct and indirect mechanisms, as the factor governing the maintenance of the neurosecretory process and the properties of dense-core vesicles in PC12 cells.


Cyfluthrin (EW 050)-impregnated bednets in a malaria control program in Ghassreghand (Baluchistan, Iran).
In a study carried out in the Ghassreghand Division (Baluchistan, Iran) from March through November 1995, efficacy of cyfluthrin-impregnated bednets was compared to that of untreated nets, in relation to malaria control. Ten villages with a total population of 4,572 and 3 villages with a total population of 1,935 were used as treatment and control, respectively. The collection, impregnation (target dosage of 40 mg active ingredient [AI]/m2), and redistribution of the nets (9% nylon, 52% light cotton, 30% medium cotton, and 9% heavy cotton), carried out in mid-April, were done by local health workers, supervised by the senior research staff. Anopheles culicifacies was considered to be the main vector of malaria in the named area. This species is mainly zoophilic, endophilic, and exophagic. The initial uptake of the insecticide was lower than the target dosage, with high variation (nylon, 12.5 +/- 5.4 mg AI/m2; light cotton, 33.3 +/- 26.1 mg AI/m2; medium cotton, 25.9 +/- 20 mg AI/m2; heavy cotton, 17.6 +/- 12.5 mg AI/m2). The use of impregnated mosquito nets (used primarily outside) had no significant effect on the incidence of malaria. No difference was detected in the parasite density of patients with positive slides. No significant effect was observed in the parous rate, human blood index, and sporozoite rate of anopheline vectors. Only the indoor resting densities of An. culicifacies and other malaria vectors were drastically reduced after the introduction of the cyfluthrin-impregnated nets into the treatment villages. The residual activity of cyfluthrin was lower than expected. The mortality of anophelines brought in contact with the treated nets for 3 min in bioassays dropped to less than 55% in 3 months. The loss of chemical activity was greatest for the light cotton nets, followed by the medium cotton nets. Cyfluthrin-treated nets were mildly irritating to host-seeking female anophelines in the laboratory. The protective rate of impregnation (all fabric kinds included) in preventing female mosquitoes from biting through the impregnated nets was initially 5-6 times that of the nonimpregnated nets. The study did not detect any significant difference between the use of untreated versus impregnated bednets in the Ghassreghand area. In planning future medium-scale trials, comparison of new compounds and formulations to the more widely used pyrethroids such as permethrin and deltamethrin is highly recommended.


Use of a Pressure Cooker to Achieve Sterilization for an Expeditionary Environment.
Sterilization of healthcare instruments in an expeditionary environment presents a myriad of challenges including portability, cost, and sufficient electrical power. Using pressure cookers to sterilize instruments presents a low-cost option for sterilization in prehospital settings. This project's objective was to determine if sterility can be achieved using a commercially available pressure cooker. Presto® 4-quart stainless steel pressure cookers were heated using Cuisinart® CB-30 cast-iron single burners. One 3M™ Attest™ 1292 Rapid Readout Biological Indicator and one 3M™ Comply™ SteriGage™ integrator strip were sealed in a Henry Schein® Sterilization Pouch and placed in a pressure cooker and brought to a pressure of 103.4kPa. Sterility was verified after 20 minutes at pressure. The Attest vials were incubated in a 3M Attest 290 Auto-Reader for 3 hours with a control vial. Sterility using the pressure cooker was achieved in all tested bags, integrator strips, and Attest vials (n = 128). The mean time to achieve the necessary 103.4kPa was 379 seconds (standard deviation (SD) = 77). Neither the ambient temperature nor humidity were found to affect the pressure cooker's time to achieve adequate pressure, nor the achieved depth on the integrator strip (all p > .05). This study provides evidence that sterilization is possible with offthe- shelf pressure cookers. Though lacking US Food and Drug Administration (FDA) approval, the use of this commercially available pressure cooker may provide a method of sterilization requiring minimal resources from providers working in expeditionary environments.


[Nutrition characteristics and ascorbic acid requirements in students living in an arid zone].
Data are presented on the actual nutrition and providing with vitamin C in students of the V. I. Lenin Pedagogical Institute (Ashkhabad, Turkmenia). Twenty male students aged 18-24 years were investigated during the examination session in summer. Group 1 (12 subjects) received an additional dose of ascorbic acid (150 mg/day), group 2 (8 subjects) served as control. Diversion from the principles of the rational nutrition was established in the actual nutrition of the students investigated: in summer, due to low appetite, students eat low amounts of meat products, that results in the decreased content of animal proteins and fats in their daily rations as compared to the values recommended for this group of population. Vitamin C deficiency was also detected in the students studied. Additional intake of ascorbic acid in a dose of 150 mg/day has normalized their providing with this vitamin.


Relationship between abdominal aortic and coronary artery calcification as detected by computed tomography in chronic kidney disease patients.
The purpose of this study was to investigate the relationship between abdominal aortic calcification (AAC) and coronary artery calcification (CAC) in chronic kidney disease (CKD) patients. We evaluated 126 asymptomatic CKD patients (mean estimated glomerular filtration rate: 36.1 ± 14.1 mL/min/1.73 m(2), mean age 70.3 ± 10.1 years). A non-contrast computed tomography scan was used to determine the abdominal aortic calcification index (ACI) and CAC score, and this relationship was investigated. Among the subjects, AAC was present in 109 patients (86.5 %) as defined by ACI >0 and median ACI was 11.7 %. ACI increased in accordance with advances in CAC score grades (3.0, 5.2, 17.2, and 32.8 % for CAC score 0, 1-100, 101-400, and 401 or more, respectively, p < 0.001). Even after multivariate adjustment, ACI was independently associated with severe CAC score as defined by CAC score >400 [odds ratio 1.08, 95 % confidence interval (CI) 1.04-1.12, p < 0.001]. Receiver-operating curve analysis showed that the ACI optimal cut-off value predicting severe CAC score was 16.5 % (area under the curve = 0.79, 95 % CI 0.69-0.90, p < 0.001). The C statics for predicting CAC score was significantly increased by adding ACI values to the model including other risk factors (0.853 versus 0.737, p = 0.023). In conclusion, the ACI value of 16.5 % allows us to predict the presence of severe CAC in CKD patients, and that the addition of ACI to the model with traditional risk factors significantly improves the predictive ability of severe CAC score. These data reinforce the utility of ACI as a screening tool in clinical practice.


Perirenal fat diameter measured by echography could be an early predictor of lipodystrophy in HIV type 1-infected patients receiving highly active antiretroviral therapy.
Echographically measured thicknesses of perirenal and subcutaneous fat, as well as serum metabolic and anthropometric parameters, were evaluated in 74 human immunodeficiency virus-infected patients receiving highly active antiretroviral therapy (HAART), 22 of whom were HAART-naive at baseline, who were followed-up for 27 months to detect predictive factors of lipodystrophy. Perirenal fat diameter (PRFD) at baseline differed in HAART-naive and HAART-experienced patients (P<.001), and it was the best predictor of lipodystrophy changes after 12 months of follow-up in the HAART-naive patients (hazard ratio, 7.34; 95% confidence interval, 1.18-45.49; P=.032). In addition, HAART-experienced patients in whom lipodystrophy improved had thinner baseline perirenal fat than those in whom lipodystrophy did not improve (P=.04). A PRFD of >2.6 mm at baseline or >4.9 mm during receipt of HAART suggested lipodystrophy predisposition. PRFD correlated significantly with other metabolic and anthropometric parameters. Echographically measured PRFD is associated with lipodystrophy and could be used as an early predictor of this syndrome in treatment-naive patients starting HAART.


Does change in positive affect mediate and/or moderate the impact of symptom distress on psychological adjustment after cancer diagnosis? A prospective analysis.
Physical symptom distress is one of the commonest correlates of psychological adjustment in cancer patients. Positive affect (PA) can be a dynamic resource for patients to cope with the cancer-related physical demands. The present study examined whether differential patterns of change in PA were associated with anxiety and depressed mood, and whether PA modified the association between change in symptom distress and psychological distress in 215 Chinese people newly diagnosed with colorectal cancer (CRC). Participants completed measures of physical symptoms, PA, and anxiety and depression at diagnosis and again at 3-month follow-up. Multivariate analyses of covariance revealed that at follow-up, people reporting higher anxiety and depressed mood demonstrated loss in PA, whereas those reporting lower depressed mood demonstrated a gain in PA. Structural equation modelling revealed that change in PA significantly mediated and moderated the associations between increased symptom distress and anxiety and depressed mood. We conclude that in line with Hobfoll's conservation of resources theory, continuous physical symptom distress depletes PA of newly diagnosed cancer patients, resulting in higher levels of anxiety and depressed mood. Effectiveness of symptom management intervention could be enhanced by preserving or enhancing PA in patients.


More than Taking a Chair: The Perceived Group Social Dynamics of Alcoholics Anonymous Related to Changes in Spiritual Practices.
Alcoholics Anonymous (AA) meeting attendance and spiritual practices are established predictors of abstinence. This study utilized longitudinal data from two studies of AA to investigate 1) how perceived within meeting social dynamics in AA meetings affect later AA attendance, and thus exposure to the emphasis of spiritual practices and 2) influence the extent that spiritual gains are mobilized, beyond AA attendance. Findings revealed that greater group cohesion was associated with lower AA attrition, and that expressiveness, or openness, of the group predicted reported practice of spiritual behaviors. Findings reveal distinct group dynamics may foster distinct mechanism of change of AA attendees.


Biochemical characterization of RNA coliphage MX1.
In order to clarify the taxonomic status of an RNA coliphage, MX1 (a serological intermediate between groups III and IV), we examined (i) read-through protein synthesis in a cell-free protein-synthesizing system, (ii) the peptide map of the coat protein and (iii) the RNA sequence in the 3'-terminal region of MX1 RNA. We found that the characteristics of MX1 were closer to those of group III phages than to those of group IV. For example, the gel elecrophoretic pattern of the protein coded for by MX1 RNA was the same as that of group III phage proteins. Peptide fingerprints of the coat protein of MX1 showed that seven tryptic peptides overlapped with corresponding peptides of Q beta (group III phage), whereas only two peptides overlapped with those of SP (group IV phage). Furthermore, in the base sequence of the first 200 nucleotides from the 3'-end of MX1 RNA, about 70% of the nucleotides were homologous to those of the Q beta RNA, whereas the homology to SP RNA was only 53%. These results suggest that MX1 is more closely related to group III phages than to group IV phages and we propose that it should be assigned to the former group.


Clonal dynamics limits detection of selection in tumour xenograft CRISPR/Cas9 screens.
Transplantable in vivo CRISPR/Cas9 knockout screens, in which cells are edited in vitro and inoculated into mice to form tumours, allow evaluation of gene function in a cancer model that incorporates the multicellular interactions of the tumour microenvironment. To improve our understanding of the key parameters for success with this method, we investigated the choice of cell line, mouse host, tumour harvesting timepoint and guide RNA (gRNA) library size. We found that high gRNA (80-95%) representation was maintained in a HCT116 subline transduced with the GeCKOv2 whole-genome gRNA library and transplanted into NSG mice when tumours were harvested at early (14 d) but not late time points (38-43 d). The decreased representation in older tumours was accompanied by large increases in variance in gRNA read counts, with notable expansion of a small number of random clones in each sample. The variable clonal dynamics resulted in a high level of 'noise' that limited the detection of gRNA-based selection. Using simulated datasets derived from our experimental data, we show that considerable reductions in count variance would be achieved with smaller library sizes. Based on our findings, we suggest a pathway to rationally design adequately powered in vivo CRISPR screens for successful evaluation of gene function.


Detection of Haemophilus influenzae type B DNA in a murine pneumonia model by in situ PCR.
This study estimated the value of in situ PCR (ISPCR) in the detection of Haemophilus influenzae type b (Hib) DNA in paraffin-embedded lung tissues of a murine pneumonia model. ICR mice were infected with Hib solution intranasally. In study group A (n=20), physiological changes and the number of deaths were recorded for 7 consecutive days after infection. In study group B (n=10), blood samples and lung tissues were obtained from the infected mice on the brink of death. In both groups, portions of the lung tissue were cultured for Hib, while other portions were submitted for histopathological studies. Conventional PCR, PCR followed by Southern blotting and ISPCR were performed to detect Hib in paraffin-embedded lung tissues. In control group A, six mice were inoculated intranasally with the same concentration of heat-inactivated Hib solution. In control group B, six healthy mice served as a blank control. Both control groups were managed using the same methods as those used in the study groups. The white blood cell count of the mice in the study group increased (F=3.295, P<0.01), with a high neutrophil count (F=0.127, P<0.05). In the histopathological study, various stages of pneumonia were found in the lung tissues of the infected mice examined by microscope; 80% of the mice had moderate or severe pneumonia. Cultures of lung tissues in the study groups were all positive for Hib, while no bacteria were found in the control groups. Hib was detected in only 4 of 30 samples (13.3%) of the study groups using conventional PCR, but in all 30 samples (100%) using both Southern blotting and ISPCR. All three methods did not detect Hib in the control groups. Because of its sensitivity and specificity and its ability to locate the micro-organism, ISPCR can be considered suitable for the detection of Hib in paraffin-embedded lung tissues.


Skull Base Tumors.
This article reviews the presenting features, molecular characteristics, diagnosis, and management of selected skull base tumors, including meningiomas, vestibular schwannomas, pituitary neuroendocrine tumors, craniopharyngiomas, chordomas, ecchordosis physaliphora, chondrosarcomas, esthesioneuroblastomas, and paragangliomas. Skull base tumors pose a management challenge given their complex location and, as a result, the tumors and treatment can result in significant morbidity. In most cases, surgery, radiation therapy, or both yield high rates of disease control, but the use of these therapies may be limited by the surgical accessibility of these tumors and their proximity to critical structures. The World Health Organization classification of pituitary neuroendocrine tumors was updated in 2022. Scientific advances have led to an enhanced understanding of the genetic drivers of many types of skull base tumors and have revealed several potentially targetable genetic alterations. This information is being leveraged in the design of ongoing clinical trials, with the hope of rendering these challenging tumors treatable through less invasive and morbid measures. Tumors involving the skull base are heterogeneous and may arise from bony structures, cranial nerves, the meninges, the sinonasal tract, the pituitary gland, or embryonic tissues. Treatment often requires a multidisciplinary approach, with participation from radiation oncologists, medical oncologists, neuro-oncologists, and surgical specialists, including neurosurgeons, otolaryngologists, and head and neck surgeons. Treatment has largely centered around surgical resection, when feasible, and the use of first-line or salvage radiation therapy, with chemotherapy, targeted therapy, or both considered in selected settings. Our growing understanding of the molecular drivers of these diseases may facilitate future expansion of pharmacologic options to treat skull base tumors.


Pneumocystis carinii exhibits a conserved meiotic control pathway.
Pneumocystis carinii is an opportunistic fungus causing severe pneumonia in immune-compromised hosts. Recent evidence suggests that Pneumocystis exists as separate sex types, though definitive evidence is currently lacking. These studies were undertaken to determine whether Pneumocystis maintains functional meiotic control molecules, which are required for sexual life stages in eukaryotes. Using the Pneumocystis carinii Genome Project database, two partial sequences for meiotic control molecules were detected, namely, PCRan1, a presumptive meiotic control kinase, and PCMei2, a homologue to a primary activator of meiosis in Schizosaccharomyces pombe. Rapid amplification of cDNA ends was employed to obtain the full open reading frames and to further investigate the functions of these proteins. These presumptive meiotic control molecules were most homologous to molecules present in S. pombe (52% identical and 67% homologous for PCRan1 and 75% identical and 88% homologous for PCMei2 by BLAST analysis). Heterologous expression of these Pneumocystis meiotic genes in corresponding temperature-sensitive and knockout strains of S. pombe, respectively, further verified the functions of the PCRan1 and PCMei2 proteins. These proteins were further shown to control downstream components of the meiotic pathway in S. pombe. Lastly, in vitro kinase assays were used to determine that PCRan1p phosphorylates PCMei2p. These experiments represent the first characterization of any proteins in P. carinii involved in meiosis and indicate the presence of a conserved meiotic pathway in Pneumocystis. Elucidation of this pathway will be essential in gaining a greater understanding of this important opportunistic fungal pathogen.


[Glutamatergic mechanisms in alcohol dependence--genetic, molecular-biological and neuropharmacological findings].
A number of preclinical and clinical findings suggest that the glutamate system, especially NMDA-receptors has a major function in neuropsychiatric disorders related to chronic alcoholism. Other findings suggest, that glutamatergic mechanisms may also play a role for the addictive process itself including craving and addiction memory. A number of genetic and molecular-biological findings and more recent neuropharmacological studies point in this direction. NMDA-modulators and antagonists have been found to be possible or even clinically effective so called anti-craving-drugs. A better understanding of the glutamatergic mechanisms in alcohol dependence may therefore also be of relevance for the development of new anti-craving-drugs and therapy-research.


False positive seroreactivity to brucellosis in tuberculosis patients: a prevalence study.
The rising worldwide incidence of tuberculosis (TB) increases the demand for knowledge about its potential seroreactivity with other microbial agents. A few reports and the authors' experiences indicate that tuberculosis may result in a false-positive brucellosis serology. This may cause a diagnostic challenge because of the close clinical resemblance of these two infections. The aim of the present prevalence study was to elucidate brucellosis seroreactivity in patients with active TB. Ninety-eight patients with newly diagnosed and active TB were studied using an enzyme-linked immunosorbent assay (ELISA) and Wright's and Coombs-Wright's tests. Seventy-five healthy individuals were used as controls. The patients showed signs of recovery after starting a standard anti-TB regimen and had no clinical evidence of brucellosis at a subsequent 6-month follow-up. The data were analyzed statistically by Fisher's exact test using SPSS 11.0. We found that 9.2% of TB patients versus 1.3% of healthy controls had positive results on the anti-Brucella IgG ELISA (P = 0.04). Five TB patients were found to have agglutination on Wright's tests, while none of the controls showed agglutination. Active TB patients may have some seroreactivity with Brucella antigens, and Brucella IgG ELISA may give a false positive in these patients. Clinicians should consider false positive brucellosis seroreactivity in patients with active TB.


Geo-epidemiologic mapping in the new public health surveillance. The malaria case in Chiapas, Mexico, 2002.
The new public health surveillance requires at the global, national and local levels the use of new authoritative analytical approaches and tools for better recognition of the epidemiologic characteristics of the priority health events and risk factors affecting the population health. The identification of the events in time and space is of fundamental importance so that the geo-spatial description of the situation of diseases and health events facilitates the identification of social, environmental and health care related risks. This assessment examines the application and use of geo-spatial tools for identifying relevant spatial and epidemiological conglomerates of malaria in Chiapas, Mexico. The study design was ecological and the level of aggregation of the collected information of the epidemiological and spatial variables was municipalities. The data were collected in all municipalities of the state of Chiapas, Mexico during the years 2000-2002. The main outcome variable was cases and types of malaria diagnosed by blood smears in weekly reports. Independent variables were age, sex, ethnicity, literacy of the cases of malaria and environmental factors such as altitude, road type and network in the municipalities and cities of Chiapas. The production of thematic maps and the application of geo-spatial analytical tools such Moran and local indicator of spatial autocorrelation metrics for malaria clustering allowed the visualization and recognition that the important population risk factors associated with high malaria incidence in Chiapas were low literacy rate, areas with high percentage of indigenous population that reflects the social inequalities gaps in health and the great burden of disease that is affecting this important vulnerable group in Chiapas. The presence of road networks allowed greater spatial diffusion of Malaria. An important epidemiological and spatial cluster of malaria was identified in the areas and populations in the proximity of the southern border. The use of geospatial metrics in local areas will assist in the epidemiological stratification of malaria for better targeting more effective and equitable prevention and control interventions.


Syphilis epidemiology in 1994-2013, molecular epidemiological strain typing and determination of macrolide resistance in Treponema pallidum in 2013-2014 in Tuva Republic, Russia.
The incidence of syphilis in the Tuva Republic (geographical centre of Asia), Russia has been exceedingly high historically. No detailed examinations and no molecular investigations of Treponema pallidum strains transmitted in the Tuva Republic, or in general, in Russia, were published internationally. We examined the syphilis epidemiology in 1994-2013, and the molecular epidemiology and macrolide resistance in T. pallidum strains in 2013-2014 in the Tuva Republic. Among 95 mainly primary or secondary syphilis patients, the arp, tpr, tp0548 and 23S rRNA genes in 85 polA gene-positive genital ulcer specimens were characterized. The syphilis incidence in Tuva Republic peaked in 1998 (1562), however declined to 177 in 2013. Among the 70 (82%) completely genotyped specimens, six molecular strain types were found. Strain type 14d/f accounted for 91%, but also 14c/f, 14d/g, 14b/f, 14i/f, 9d/f, and 4d/f were identified. Two (2.4%) specimens contained the 23S rRNA A2058G macrolide resistance mutation. This is the first internationally published typing study regarding T. pallidum in Russia, performed in the Tuva Republic with the highest syphilis incidence in Russia. The two molecular strain types 4d/f and 9d/f have previously been described only in Eastern and Northern China and for the first time, macrolide-resistant syphilis was described in Russia.


Tourniquet use in ankle arthroscopy: A systematic review.
Ankle arthroscopy is commonly performed using a thigh tourniquet and is thought to improve visibility and reduce operative time. However, the current evidence is unclear as to whether the use of a tourniquet provides these benefits. The aim of this study was to investigate whether there is any clinical benefit of using a tourniquet in ankle arthroscopy. A systematic review following PRISMA guidelines was undertaken. All clinical studies published in Medline, Embase, PubMed and the Cochrane Library Database from inception until January 2023 reporting on the use of a tourniquet in ankle arthroscopy were included. 180 studies were identified of which 3 (164 patients) met the inclusion criteria. All studies showed no statistically significant difference in mean surgical time and complication rate between the tourniquet and non-tourniquet groups. Overall, the quality of the evidence was moderate to poor without data in favour or against the routine use of tourniquets in ankle arthroscopy. The current literature suggests that there are no significant differences in mean surgical time and complication rate between the tourniquet and non-tourniquet groups.


[Medical physics. Perspectives of organizing a new specialty].
A case is made for setting up a new certificate program "Medical Physics". This would take care of specialists certified in physics and applied technological disciplines who work in hospitals and, particularly, radiological wards. The Association of Medical Physicists of Russia, established in 1991, is making a considerable contribution to the training of medical physicists in full conformity with recommendations of international organization and agencies.


Tactile stimulation reduces fear in fish.
Being groomed or touched can counter stress and negative affect in mammals. In two experiments we explored whether a similar phenomenon exists in non-mammals like zebrafish. In Experiment 1, we exposed zebrafish to a natural stressor, a chemical alarm signal released by injured conspecifics. Before moving them into an observation tank, one group of fish was washed and then subjected to a water current that served as the tactile stimulus. The other group was simply washed. Fish with tactile treatment demonstrated fewer fear behaviors (e.g., bottom dwelling) and lower cortisol levels than fish without. In Experiment 2, we ascertained a role of somatosensation in these effects. Using a similar paradigm as in Experiment 1, we recorded fear behaviors of intact fish and fish with damaged lateral line hair cells. Relative to the former, the latter benefited less from the tactile stimulus during fear recovery. Together these findings show that tactile stimulation can calm fish and that tactile receptors, evolutionarily older than those present in mammals, contribute to this phenomenon.


Determining the foraging range and origin of resurgence after treatment of Argentine ant (Hymenoptera: Formicidae) in urban areas.
The foraging range and distribution of Argentine ant, Linepithema humile (Mayr), colonies in urban areas of southern California extended at least 61 m (200 feet) from feeding stations and structures. Ants were fed at 25% sucrose feeding stations containing 0.01% fluorescent brighter (FB28). Within 14 d, from 77-90% of the ants sampled next to the feeding stations were positive for FB28. The percentage of ants with FB28 declined gradually to approximately 55% 61 m away from the feeding station. The percentage of marked ants in the controls didn't change over the 4-wk-test period. There were approximately 290,000 ants visiting the monitored stations each night before treatments. The 0.0001% fipronil baits and 0.06% fipronil SC sprays provided significant reductions in at least 4 wk. The percentage of ants marked with FB28 decreased significantly in both bait and spray treatments over 4 wk suggesting that the resurgence of ants in treated areas were because of immigration from untreated areas. It is likely that much larger areas will need to be treated to control Argentine ants in urban settings, especially if baits are being used.


Effect of sodium selenite and selenium yeast on performance, egg quality, antioxidant capacity, and selenium deposition of laying hens.
This study compared the effects of sodium selenite and selenium yeast and their combination on laying performance, egg quality, antioxidant capacity, and selenium (Se) contents in tissues and eggs. Two-hundred-eighty-eight Jing Hong layers that were similar in laying rate (87.5 ± 0.38%) and body weight (1.70 ± 0.02 kg) were randomly distributed into 4 treatments for 11 wk (from 203 d old to 279 d old) with 9 replicates of 8 hens per replicate. The diets (corn-soybean meal diet) were supplemented with 0 [blank control (BC)], 0.3 mg/kg Se from sodium selenite (SS), 0.15 mg/kg Se from sodium selenite and 0.15 mg/kg Se from Se yeast (SS+SY), or 0.3 mg/kg Se from Se yeast (SY). Results showed that the laying rate of the SS+SY group increased significantly (P < 0.05) compared to the BC and SY groups. There were no differences (P > 0.05) in egg quality between the Se-supplemented diets and the BC diet. The serum glutathione peroxidase (GSH-Px) activity was increased (P < 0.01) in hens fed Se-supplemented diets compared to the BC diet. The liver superoxide dismutase (SOD) activity of the SY group was increased significantly (P < 0.05) compared to the BC group. Significant increase (P < 0.01) due to SY supplementation was noted in the serum vitamin E content compared to BC and SS. Layers fed Se-supplemented diets had higher (P < 0.01) contents of Se in the serum, liver, and kidney compared to the BC diet. Compared to BC, Se content in eggs was significantly increased (P < 0.05) by feeding supplementary Se. In conclusion, the effects of SS and Se yeast were approximately equal in promoting antioxidant capacity of laying hens, while Se yeast is easier to deposit into eggs and tissues. The diet with added equal amounts of the 2 sources of Se was more cost effective and affordable than a comparable amount of Se yeast to obtain the promising production performance and nearly similar Se deposition.


[Additive therapy in ovulation induction with clomiphene].
Between 1977 and 1981 1319 patients consulted our hospital with the problem of sterility. 763 women became pregnant, 200 were treated with clomiphen . In 69 cases we used a combination with other agents, because ovulation could not be induced with clomiphen alone. 7 times we combined clomiphen with HCG, in 7 other cases with Prednisolon . Ovulation and pregnancy occurred subsequent to a treatment with Estradiolvalerianate -Norgestrel and clomiphen in 15 patients, in 32 patients after administration of a combination of clomiphen and Bromocriptine. The inadequate cervical mucus which may develop in patients during clomiphen therapy was treated with an additional low dose of ethinylestradiol. Our retrospective study confirms the value of a combination therapy in the treatment of anovulation and sterility.


Use of inhaled corticosteroids and bone mineral density in a population based study: the Nord-Trøndelag Health Study (the HUNT Study).
Conflicting results have been reported of the long-term effects of treatment with inhaled corticosteroids (ICS) on bone. The objective of this study was to compare ICS users and non-users regarding bone mineral density (BMD) in a large population. A total of 65,225 adults participated in a cross-sectional study in the Nord-Trøndelag Health Study 1995-1997. Those reporting asthma or asthma-related symptoms, were invited to have bone densitometry of the forearm, flow volume spirometry and a personal interview. Altogether 4482 women and 4142 men participated, of whom 2113 reported ever use and 6511 never use of ICS. Never-users of corticosteroids had a mean BMD, adjusted for confounders (age, square age, sex, body mass index, height, physical activity, work load, packyears, family history of osteoporosis and in women number of years since menopause and use of hormone replacement therapy), of 0.493 g/cm2 at the distal site. Subjects having only used ICS or combined with courses of prednisolone, had 0.008 g/cm2 (95%CI: 0.005-0.011) lower BMD whilst users of prednisolone > or = 6 months had 0.038 g/cm2 (0.021-0.055) lower level. No dose response association between ICS and BMD, or difference in BMD by type of ICS was found. The association between CS use and BMD was independent of the measuring site. ICS use was associated with lower BMD. The lack of dose response in this study might be due to a narrow dose range or indicates that other characteristics of the patient group are contributing to the observed difference in ICS users compared to never-users.


Expression of metallopanstimulin and oncogenesis in human prostatic carcinoma.
We have previously shown that human metallopanstimulin (MPS-1) is a 9.4-kDa multifunctional ribosomal S27/nuclear "zinc finger" protein which is expressed in a wide variety of actively proliferating cells and tumor tissues. Furthermore, we have shown that detection of MPS-N immunoreactive material in sera corresponding to the NH2 terminus of MPS-1 provides a method for determining the presence of certain types of abnormal proliferative conditions and/or active oncogenic processes in patients. In this study, we investigated MPS-N and MPS-N-like antigens present in the blood of patients with prostatic carcinoma (PC) and their relationship to the clinical status of patients with PC. The presence of MPS-N immunoreactive material was determined using a sensitive and specific radioimmunoassay (RIA) which has been developed to measure circulating levels of MPS-N antigen(s). In addition, MPS-N levels were compared to the primary bio-marker used in PC patient management, Prostatic Specific Antigen (PSA). MPS-N concentrations were determined in the blood of 107 males having no evidence of PC, and in 126 patients diagnosed with PC. In patients, not having PC the MPS-N levels were lower than 10 ng/mL. In untreated patients having PC stages T1/T2, the MPS-N level range was 10-30 ng/mL; in stages T3/T4 the MPS-N level range was 30-50 ng/mL; and in stage Mlb (distant metastasis to the bones) the MPS-N levels were extremely high (> 50 ng/mL). In Mlb patients that did not respond to therapy, the MPS-N levels remained very high (> 50 ng/mL). In Mlb patients that went into remission after treatment, the MPS-N levels were dramatically reduced. In addition, a comparison of the test properties of MPS-N and PSA for prostate cancer were evaluated in a total of 231 patients. In both the low and high value range, both MPS-N and PSA appear to be equally effective in modifying the probability of the target condition-prostatic cancer. These findings show that (1) in untreated PC patients, the increase in serum MPS-N correlated with the stage of the disease; (2) MPS-N tumor marker predicted the degree of aggressiveness of tumor growth and response to therapy. In summary, despite the uncertainties of the relative contributions of the molecules being measured in cancer patients (authentic MPS-1, and MPS-N-like protein sequences), the MPS-N test is a pragmatic test that correlates well with active prostatic malignancy.


Effect of alcohol on the heart and cardiac metabolism.
In this report the disturbances in biochemistry of the heart muscle exposed to alcohol are delineated. All elements of cellular substructures are affected. In plasma membranes, (Na+ + K+)-activated ATPase (EC 22.214.171.124) is inhibited. Mitochondrial damage consists in diminished respiratory function and calcium uptake and binding. High-energy phosphates remain intact. Alcohol also affects the malate-aspartate shuttle. Acetaldehyde, a metabolite of ethanol, has a direct effect on myocardial protein synthesis through microsomal inhibition; however, the development of cardiac hypertrophy is not affected. Malfunction of sarcoplasmic reticulum is evidenced by disturbances in calcium binding and uptake. Effects of ethanol on the contractile machinery are deficiencies in the turnover rate of chemical into mechanical energy (diminished Vmax), and in the number of cross-bridges formed (P0). It increases stiffness of series elastic elements. There is diminished fatty acid oxidation with increased esterification. The involvement of CoA synthetase (EC 220.127.116.11), palmityl-carnitine transferase (EC 220.127.116.11), and pyruvate dehydrogenase complex in disturbed fatty acid oxidation is not certain. The role of catalase in myocardial ethanol oxidation was examined. Ethanol activates myocardial catalase-H2O2 complex (EC 220.127.116.11). The biochemical basis of fetal alcohol syndrome is low hepatic alcohol dehydrogenase (EC 188.8.131.52) activity during fetal life.


Influence of a new fluoroquinolone, AF3013 (the active metabolite of prulifloxacin), on macrophage functions against Klebsiella pneumoniae: an in vitro comparison with pefloxacin.
The efficacy of an antibiotic in the treatment of bacterial infections depends upon the interaction of bacterium, drug and phagocytes. In this study we have investigated the influence of AF3013, a new fluoroquinolone, on the activities of mouse peritoneal macrophages against Klebsiella pneumoniae, in comparison with the influence of pefloxacin. Bacterial susceptibility to phagocytosis and intracellular killing were determined after klebsiellae and macrophages had been incubated simultaneously with inhibitory concentrations of both AF3013 and pefloxacin and following pre-exposure of the microorganisms and the macrophages individually to the same concentrations of each drug. Under the experimental conditions used, both AF3013 and pefloxacin potentiated the phagocytic and microbicidal activities of the macrophages, although different mechanisms may be involved.


Dominant Polar Surfaces of Colloidal II-VI Wurtzite Semiconductor Nanocrystals Enabled by Cation Exchange.
Polar surfaces of ionic crystals are of growing technological importance, with implications for the efficiency of photocatalysts, gas sensors, and electronic devices. The creation of ionic nanocrystals with high percentages of polar surfaces is an option for improving their efficiency in the aforementioned applications but is hard to accomplish because they are less thermodynamically stable and prone to vanish during the growth process. Herein, we develop a strategy that is capable of producing polar surface-dominated II-VI semiconductor nanocrystals, including ZnS and CdS, from copper sulfide hexagonal nanoplates through cation exchange reactions. The obtained wurtzite ZnS hexagonal nanoplates have dominant {002} polar surfaces, occupying up to 97.8% of all surfaces. Density functional theory calculations reveal the polar surfaces can be stabilized by a charge transfer of 0.25 eV/formula from the anion-terminated surface to the cation-terminated surface, which also explains the presence of polar surfaces in the initial Cu1.75S hexagonal nanoplates with cation deficiency prior to cation exchange reactions. Experimental results showed that the HER activity could be boosted by the surface polarization of polar surface-dominated ZnS hexagonal nanoplates. We anticipate this strategy is general and could be used with other systems to prepare nanocrystals with dominant polar surfaces. Furthermore, the availability of colloidal semiconductor nanocrystals with dominant polar surfaces produced through this strategy opens a new avenue for improving their efficiency in catalysis, photocatalysis, gas sensing, and other applications.


The relative sensitivity of four benthic invertebrates to metals in spiked-sediment exposures and application to contaminated field sediment.
The relative sensitivity of four benthic invertebrates (Hyalella azteca, Chironomus riparius, Hexagenia spp., and Tubifex tubifex) was determined for Cd, Cu, and Ni in water-only and in spiked-sediment exposures. Survival (median lethal concentrations [LC50s] and the concentrations estimated to be lethal to 25% of test organisms [LC25s]), and endpoints for growth and reproduction (mean inhibitory concentrations [IC25s]) were compared. The sensitivities differed depending on the species and metal, although some trends emerged. In water-only exposures, H. azteca is the most sensitive species to cadmium and nickel, with mean LC50s of 0.013 and 3.6 mg/L, respectively; C. riparius is the most sensitive species to copper, with a mean LC50 of 0.043 mg/L. In the spiked-sediment exposures, the order in decreasing sensitivity to copper is Hyalella = Hexagenia < Chironomus < Tubifex for survival and growth/reproduction. For cadmium, the order in decreasing sensitivity is Hyalella = Chironomus < Hexagenia < Tubifex, and for nickel is Hyalella << Hexagenia < Chironomus < Tubifex. Chironomus riparius and Hexagenia spp. survival can be used to distinguish between toxicity caused by different metals. Species test responses in field-collected sediment(Collingwood Harbour, ON, Canada) were examined in an attempt to determine the causative agent of toxicity throughout, using the established species sensitivities. Sediment toxicity was categorized first by comparing species responses to those established for a reference database. Test responses in the field-collected sediment do not support causality by Cu, a suspected toxicant based on comparison of sediment chemistry with sediment quality guidelines.


Non-linear assessment of anticancer activity of 17-picolyl and 17-picolinylidene androstane derivatives--chemometric guidelines for further syntheses.
The present paper deals with prediction of cytotoxic activity of 17-picolyl and 17-picolinylidene androstane derivatives toward androgen receptor negative prostate cancer cell line (PC-3). The prediction was achieved applying artificial neural networks (ANNs) method on the basis of molecular descriptors. The most important descriptors (skin permeability (SP), Madin-Darby canine kidney cell permeability (MDCK) and universal salt solubility factor (S+SF)) were selected by using stepwise selection coupled with partial least squares method. The ANN modelling was carried out in order to obtain reliable models which can facilitate further synthesis of androstane derivatives with high antiproliferative activity toward PC-3 cell line. The modelling procedure resulted in three ANN models with the best statistical performance. The obtained results show that the established ANN models can be applied for required purpose.


[Legislation on smoking at workplace and in public places in Europe].
This study was conducted through contacts with key health authorities, complemented with other key providers of information and the consultation of the International Healthcare Legislation Repertory. A description is provided of the current characteristics of the legislation on smoking at the workplace in the fifteen European Union (EU) Member States., as well as in other European countries such as Hungary, Iceland, Norway, Poland, Rumania and Switzerland. A review is also provided of the court cases which have taken place based upon this legislation with regard to smoking at the workplace in these countries. Legislation is a crucial part of any strategy for controlling smoking. Without legislation, no action against exposure to tobacco smoke-polluted air will be effective. However, a law does not suffice in itself and will solely be effective if supported by public opinion.


Recombinant factor VIII in the management of hemophilia A: current use and future promise.
Hemophilia A is a rare inherited bleeding disorder due to mutation of the gene that encodes the coagulation protein factor VIII. Historically, prior to the availability of treatment with factor VIII preparations, most boys died from uncontrolled bleeding, either spontaneous bleeding or after injury, before reaching 20 years of age. One of the most impressive triumphs of modern medicine is that with current recombinant factor VIII replacement therapy, a boy born in the 21st century with severe hemophilia A can anticipate a normal life expectancy with essentially no permanent complications from bleeding. For severe hemophilia A, current optimal treatment should have two goals: first, to provide sufficient factor VIII to prevent spontaneous bleeding, and second, to provide sufficient factor VIII to have normal coagulation function after any trauma. However, the replacement therapy requires tremendous resources for effective use, and remains extraordinarily expensive. Thus there are opportunities for further advances in therapy for hemophilia A. Two major concerns continue to trouble current optimal treatment approaches: some patients will develop neutralizing antibodies during the first 50 infusions of therapeutic factor VIII, and second, to administer therapeutic factor VIII every other day in young boys often requires placement of a central venous access device, and such use carries the life-threatening risks of infection and thrombosis. Because of the effectiveness of current therapy, any new developments in treatment will require significant concerns for safety, both immediate and in the long term. A number of research groups seek to prolong the biological efficacy of infused recombinant factor VIII. Currently, one such promising development is in the advanced stages of clinical trial. The goals will be to improve further the quality of life of an individual with severe hemophilia A, and to reduce the burden of current treatment strategies on families and medical resources. Hopefully, the hemophilia community will continue to participate actively in the clinical trials needed to address these new challenges.


A Nationwide Registry-Based Study on Mortality Due to Rare Congenital Anomalies.
This study aimed to analyse population-based mortality attributed to rare congenital anomalies (CAs) and assess the associated time trends and geographical differences in Spain. Data on CA-related deaths were sourced from annual mortality databases kept by the National Statistics Institute of Spain (1999⁻2013). Based on the ICD-10, only CAs corresponding to rare diseases definition were included in this study. Annual age-adjusted mortality rates were calculated and time trends were evaluated by joinpoint regression analysis. Geographical differences were assessed using standardised mortality ratios and cluster detection. A total of 13,660 rare-CA-related deaths (53.4% males) were identified in the study period. Annual age-adjusted mortality rates decreased by an average of -5.2% (-5.5% males, -4.8% females, p < 0.001). Geographical analysis showed a higher risk of rare-CA-related mortality in regions largely located in the south of the country. Despite their limitations, mortality statistics are essential and useful tools for enhancing knowledge of rare disease epidemiology and, by extension, for designing and targeting public health actions. Monitoring rare-CA-related mortality in Spain has shown a 15-year decline and geographical differences in the risk of death, all of which might well be taken into account by the health authorities in order to ensure equality and equity, and to adopt appropriate preventive measures.


Depression-focused interpersonal counseling and the use of healthcare services after myocardial infarction.
The purpose for this study was to evaluate the association between depression-focused interpersonal counseling (IPC) and the use of healthcare services in Finland after myocardial infarction (MI). The measures were done at 6 and 18 months after MI in the randomized intervention (n= 51) and the control group (n= 52). There was less use of somatic specialized healthcare services in the intervention group from 6 to 18 months after hospital discharge, and with intervention patients who had no other long-term disease during 6 months. Confirmation of possible benefits of IPC for practice calls for more specific studies.


Hyperbaric oxygen therapy of spinal cord injury.
Spinal cord injury (SCI) is a complex disease process that involves both primary and secondary mechanisms of injury and can leave patients with devastating functional impairment as well as psychological debilitation. While no curative treatment is available for spinal cord injury, current therapeutic approaches focus on reducing the secondary injury that follows SCI. Hyperbaric oxygen (HBO) therapy has shown promising neuroprotective effects in several experimental studies, but the limited number of clinical reports have shown mixed findings. This review will provide an overview of the potential mechanisms by which HBO therapy may exert neuroprotection, provide a summary of the clinical application of HBO therapy in patients with SCI, and discuss avenues for future studies.


Comparative pharmacology of the novel cyclopropylpyrroloindole-prodrug carzelesin in mice, rats, and humans.
Carzelesin is a novel cyclopropylpyrroloindole prodrug analogue that has recently been tested in Phase I clinical trials. To increase our understanding in the pharmacology of this new class of cytotoxic drugs, we have compared the pharmacology of this drug in mice, rats, and humans. The mouse was the most tolerant [10% lethal dose (LD10), 500 microg/kg], the rat was intermediate (LD10, 40 microg/kg), and humans were the least tolerant species in this series (maximum tolerated dose, 300 microg/m2 corresponding to 7.5 microg/kg). In both mice and humans, bone marrow toxicity was the primary toxic side effect. Pharmacokinetic studies, using a validated high-performance liquid chromatographic procedure, revealed that differences in drug clearance and conversion to the active drug (U-76,074) could not explain the substantial interspecies differences. The area under the plasma concentration time curve (AUCs) of carzelesin in mice and rats at their LD10s were about 80- and 20-fold higher, respectively, than in humans receiving the maximum tolerated dose, whereas the respective AUCs of U-76,074 in mice and rats were 50- and 10-fold higher. By using a colony-forming assay with bone marrow stem cells from mice and humans, we observed only a 3-fold higher toxicity in the latter. Although some of this discrepancy may be explained by the fact that the in vitro and the in vivo assays probably reflect the toxicity on different populations of colony-forming units, the tolerance of the mouse bone marrow in vivo against the very high drug levels in plasma suggest the presence of a protective mechanism, which is less active in humans. An important consequence of the much higher susceptibility of the human bone marrow for carzelesin is that the target plasma levels in humans are much below active concentrations achieved in mice, and it is clear that this may compromise the successful use of this agent in the clinic. Ultimately, however, the efficacy of this drug will be established in Phase II clinical trials.


Toxin-induced increase in survival factor receptors: modulation of the threshold for apoptosis.
The threshold at which toxins induce cell death is thought to directly relate to the amount of injury sustained. We show that the threshold at which a cell initiates toxin-induced death may vary in response to changes in the trophic environment. Treatment of Rat-1 fibroblasts with 50-175 mM dimethylformamide (DMF) induced cell death by apoptosis. Addition of insulin-like growth factor 1 (IGF-1; 100 ng/ml) and/or overexpression of the IGF-1 receptor (IGF-1R) attenuated the cytotoxicity of DMF. Furthermore, 95-99% of cells were protected from DMF-induced apoptosis if cells were pretreated with platelet-derived growth factor (5 ng/ml) for 16 h before treatment with DMF in the presence of IGF-1. Platelet-derived growth factor induced the expression of IGF-1R mRNA. The ability of cells to proliferate and survive after a 24-h treatment with DMF was determined by colony formation; whereas treatment with concentrations of >130 mM DMF reduced cellular survival, exposure to concentrations of <130 mM unexpectedly increased the colony-forming ability of treated cells when compared to that of controls. Treatment of Rat-1 fibroblasts with 75 and 130 mM DMF induced IGF-1R mRNA as determined by reverse transcription-PCR analysis. Serum withdrawal also transiently increased the expression of IGF-1R mRNA in Rat-1 fibroblasts. These results show that cells can actively adapt to pathological and physiological stress by up-regulating receptors that provide signals for cellular survival. We suggest that the threshold for toxin-induced apoptosis is determined not only by the extent of cytotoxic damage but also by the trophic environment and the ability of a cell to modulate survival signals that attenuate toxicity.