FcRγ Gene Editing Reprograms Conventional NK Cells to Display Key Features of Adaptive Human NK Cells.
Adaptive human natural killer (NK) cells display significantly enhanced responsiveness to a broad-range of antibody-bound targets through the engagement of CD16 compared to conventional NK cells, yet direct reactivity against tumor targets is generally reduced. Adaptive NK cells also display a distinct phenotype and differential expression of numerous genes, including reduced expression of signaling adapter FcRγ and transcription factor PLZF. However, it is unclear whether differential expression of specific genes is responsible for the characteristics of adaptive NK cells. Using CRISPR-Cas9, we show deletion of FcRγ in conventional NK cells led to enhanced CD16 responsiveness, abolished cell surface expression of natural cytotoxicity receptors, NKp46 and NKp30, and dramatically reduced responsiveness to K562 and Raji tumor cells. However, deletion of PLZF had no notable effects. These results suggest multiple roles for FcRγ and identify its deficiency as an important factor responsible for the functional and phenotypic characteristics exhibited by adaptive NK cells.


Incomplete type of intestinal metaplasia has the highest risk to progress to gastric cancer: results of the Spanish follow-up multicenter study.
In high or moderate risk populations, periodic surveillance of patients at risk of progression from gastric precursor lesions (PL) to gastric cancer (GC) is the most effective strategy for reducing the burden of GC. Incomplete type of intestinal metaplasia (IIM) may be considered as the best candidate, but it is still controversial and more research is needed. To further assess the progression of subtypes of IM as predictors of GC occurrence. A follow-up study was carried-out including 649 patients, diagnosed with PL between 1995-2004 in 9 participating hospitals from Spain, and who repeated the biopsy during 2011-2013. Medical information and habits were collected through a questionnaire. Based on morphology, IM was sub-classified as complete (small intestinal type, CIM) and incomplete (colonic type, IIM). Analyses were done using Cox (HR) models. At baseline, 24% of patients had atrophic gastritis, 38% CIM, 34% IIM, and 4% dysplasia. Mean follow-up was 12 years. 24 patients (3.7%) developed a gastric adenocarcinoma during follow-up. The incidence rate of GC was 2.76 and 5.76 per 1,000 person-years for those with CIM and IIM, respectively. The HR of progression to CG was 2.75 (95% CI 1.06-6.26) for those with IIM compared with those with CIM at baseline, after adjusting for sex, age, smoking, family history of GC and use of NSAIDs. IIM is the PL with highest risk to progress to GC. Sub-typing of IM is a valid procedure for the identification of high risk patients that require more intensive surveillance.


Pretreatment with interferon-γ enhances the therapeutic activity of mesenchymal stromal cells in animal models of colitis.
Mesenchymal stromal cells (MSCs) are currently under investigation for the treatment of inflammatory disorders, including Crohn's disease. MSCs are pluripotent cells with immunosuppressive properties. Recent data suggest that resting MSCs do not have significant immunomodulatory activity, but that the immunosuppressive function of MSCs has to be elicited by interferon-γ (IFN-γ). In this article, we assessed the effects of IFN-γ prestimulation of MSCs (IMSCs) on their immunosuppressive properties in vitro and in vivo. To this end, we pretreated MSCs with IFN-γ and assessed their therapeutic effects in dextran sodium sulfate (DSS)- and trinitrobenzene sulfonate (TNBS)-induced colitis in mice. We found that mice treated with IMSCs (but not MSCs) showed a significantly attenuated development of DSS-induced colitis. Furthermore, IMSCs alleviated symptoms of TNBS-induced colitis. IMSC-treated mice displayed an increase in body weight, lower colitis scores, and better survival rates compared with untreated mice. In addition, serum amyloid A protein levels and local proinflammatory cytokine levels in colonic tissues were significantly suppressed after administration of IMSC. We also observed that IMSCs showed greater migration potential than unstimulated MSCs to sites within the inflamed intestine. In conclusion, we show that prestimulation of MSCs with IFN-γ enhances their capacity to inhibit Th1 inflammatory responses, resulting in diminished mucosal damage in experimental colitis. These data demonstrate that IFN-γ activation of MSCs increases their immunosuppresive capacities and importantly, their therapeutic efficacy in vivo.


Characterization of cAMP-dependent protein kinase activation by CCK in rat pancreas.
This study reports on the use of a new sensitive assay of cAMP-dependent protein kinase activity to examine the effect of cholecystokinin (CCK) on the cAMP second messenger cascade in rat pancreatic acini. Treatment of acini with both low (pM) and high (nM) concentrations of CCK was associated with an increase in cAMP-dependent protein kinase activity. The increases in kinase activity were detected in the absence of phosphodiesterase inhibition, a condition required to detect a measurable increase in cellular cAMP in these cells. Furthermore, the cAMP cascade was dissociated from the secretory effects of CCK, since the CCK analogue, OPE, mediates enzyme secretion but does not increase cellular cAMP levels or kinase activity.


Presenilin-2 in the cynomolgus monkey brain: investigation of age-related changes.
Localization of presenilin-2 (PS-2), a transmembrane protein implicated in early onset familial Alzheimer's disease, was examined in the brains of 30 cynomolgus monkeys aged 4 to 36 years. Anti-PS-2 antibody N20, which recognizes PS-2 amino acid residues 2-20, and anti-PS-2 antibody C20, which recognizes PS-2 amino acid residues 535-554, stained mainly the cytoplasm of large pyramidal neurons and large neurites. This finding was also confirmed by double immunohistochemical investigations using N20 or C20 and anti-NeuN antibody. In the brain of the oldest monkey, swollen neurites containing senile plaques were immunostained with C20, but not with N20. Western blot analyses of microsomal fractions isolated from the brains of three adult monkeys revealed that much less PS-2 was present compared to presenilin-1 (PS-1). Age-related assessment of PS-2 in brain homogenates from young and adult monkeys showed that PS-2 levels and PS-2 subcellular localization were unchanged with increasing age. Because PS-2 expression was much less robust than that of PS-1, we conclude that PS-2 mainly localizes to large neurons and does not show so drastic age-related changes as PS-1.


Biomechanical analysis of "Barrel hoop plate" technique for the posterolateral fragments of tibial plateau fractures with different displacement tendency.
The purpose of our study is to evaluate the three-dimensional biomechanical properties of "Barrel Hoop plate" in two kinds of artificial posterolateral tibial plateau fracture fragment (PLF) by using of synthetic models, each of which has an initial amplifying displacement tendency. Thirty-six tibiae models were randomly assigned into two groups with different displacement tendencies: posterior displacement (PD) and lateral displacement (LD). Each model was then fixed with three patterns: Anterolateral plate (AP), Posterolateral plate (PP), and "Barrel Hoop plate" (BHP). Displacement in three axes of vertical, sagittal and horizontal axis was captured by Optotrak Certus motion analysis system. Bluehill 2 software was used for load control and data collection. In Model-PD, when the load was over 1000 N, the posterior displacement of Group-PP and Group-BHP were less than Group-AP (P<0.01). The inferior displacement in the vertical axis of Group-PP was larger than both Group-AP and Group-BHP in all the loading set (P<0.01). In Model-LD, both of the lateral displacement in Group-AP and Group-BHP was less than that of Group-PP when the load was over 1000 N (P<0.01). The inferior displacement of Group-AP was less than that of Group-PP in the load of 1500 N (P<0.01). Both of the posterior displacement of Group-AP and Group-BHP was less than that of Group-PP when the loading was 1500 N (P<0.01). The stiffness of Group PP was less than that of Group AP (P<0.01). The results demonstrated that the 2.7 mm "Barrel Hoop plate" had a greater capacity of anti-three-dimension axes displacement of PLF. The 3.5 mm Anterolateral plate had the advantage in anti-lateral displacement and anti-inferior displacement but was weak at anti-posterior displacement of PLF. The 2.7 mm Posterolateral plate was stronger in anti-posterior, however, weak in anti-inferior displacement capacity.


Stable Singlet Carbenes as Organic Superbases.
A simple experimental procedure for scaling carbene Brønsted basicity is described. The results highlight the strong basicity of pyrazol-4-ylidenes, a type of mesoionic carbene, also named cyclic-bentallenes (CBA). They are more basic (pKaH >42.7 in acetonitrile) than the popular proazaphosphatrane Verkade bases, and even the Schwesinger phosphazene superbase P4 (t Bu). The basicity of these compounds can readily be tuned, and they are accessible in multigram quantities. These results open new avenues for carbon centered superbases.


Ocular complications in myelodysplastic syndromes as preleukemic disorders.
To identify ocular complications in patients with myelodysplastic syndromes (MDS), who have a propensity to progress to acute myeloid leukemia (AML). Forty-one patients with MDS were the subjects in this retrospective study, and 21 patients with AML were selected as controls. Reviewing their clinical records, we verified that corneal ulcer, iridocyclitis, vitreous hemorrhage, retinal hemorrhage, and optic neuritis had been evaluated using slit-lamp assessment and opthalmoscopy in all the patients. In this study, the MDS patients were classified into those with refractory anemia (RA) and those with refractory anemia with excess blasts (RAEB). Ocular complications were found in 19 (46.3%) of the 41 patients with MDS, comprising corneal ulcer (two cases), iridocyclitis (five), vitreous hemorrhage (one), retinal hemorrhage (ten), cotton wool spots (one), and optic neuritis (two). (Some patients had more than one ocular complication.) Ocular complications were identified in 12 of the 21 (57.1%) patients with AML. There was no significant difference in frequency of ocular complications between MDS and AML (P = 0.4892). In MDS, retinal hemorrhage was associated with significantly reduced platelet counts (P = 0.0063). The frequency of ocular complications was significantly higher in MDS-RAEB than in MDS-RA (P = 0.0478). Retinal hemorrhage was significantly more frequent in patients with MDS-RAEB than in patients with MDS-RA (P = 0.0433). Ocular complications in MDS patients should be carefully examined as prognostic factors for progression to acute leukemia.


Low-molecular-weight model study of peroxide cross-linking of ethylene-propylene (-diene) rubber using gas chromatography and mass spectrometry I. Combination reactions of alkanes.
The combination reaction of linear and branched alkanes, initiated by dicumylperoxide, has been studied as a model for the combination cross-linking reaction of peroxide-cured terpolymerised ethylene, propylene and diene monomer. Both gas chromatography-mass spectrometry (GC-MS) and comprehensive two-dimensional GC-MS (GCxGC-MS) analyses have been employed to analyse the isomeric reaction products. The identification of these products based on their MS fragmentation patterns is quite complex, due to the high tendency of random rearrangements. Careful elucidation of the high-mass ions at optimised ionisation energy (55eV) has resulted in proposed structures for the different isomeric reaction products. The structure assignment by MS is in agreement with the GCxGC elution pattern and with the result of a theoretical model to predict the boiling points and, thus, the GC retention times. In addition, a model that provided a direct correlation between chemical structure and retention times was developed and this was found to provide a useful fit. Quantification of the identified reaction products by GC separation and flame ionization detection allows classification according to the hydrogen abstraction sites for the alkanes by dicumylperoxide. The selectivity for hydrogen abstraction generally follows the expected order, but a higher reactivity was observed for the methylene group next to a primary methyl group, while a reduced reactivity of the methylene group next to ethyl and to methyl groups was observed. The used approach proved to be a very powerful tool to enhance our understanding of the mechanism of peroxide cross-linking of (branched) alkanes.


[Clinical Role of Interventional Radiology in Abdominal Solid Organ Trauma].
Interventional management is commonly used for traumatic injuries to the abdominal solid organs. The American Association for the Surgery of Trauma (AAST) and the World Society of Emergency Surgery (WSES) recently published guidelines for the management and treatment of liver, spleen, and kidney injuries, emphasizing the importance of interventions. Here, we discuss the characteristics of each organ and the procedure method for each organ that interventional radiologists need to know when treating trauma patients.


An evaluation of the completeness and accuracy of active tuberculosis reporting in the United States military.
Despite the low incidence of tuberculosis (TB) in the United States military, there is uncertainty in the overall reporting and estimates of incidence. To assess TB reporting in the active component US military. TB notification in the US military was compared with three other data sources: laboratory, hospitalization and pharmacy records. Sensitivity and positive predictive value were estimated for all data sources using a gold standard of either a reportable medical event (RME) reported as confirmed or a positive laboratory result for Mycobacterium tuberculosis. Uncorrected and capture-recapture (CR) methods were used to estimate underreporting and completeness of data sources. Completeness of reporting of pulmonary TB cases was estimated as 72.4% uncorrected or 58.3% with CR. Even after correction for possible underreporting, the incidence of active pulmonary TB was only 0.87 per 100,000 person-years between 2004 and 2006. The rate of active TB in the US military is low. Like civilian surveillance, US military RME surveillance may substantially underreport TB incidence rates. Expanding surveillance to include data sources such as hospitalizations and pharmacy records will increase the number of TB diagnoses at the cost of including many false-positives.


Clinical significance of D-dimer levels during acute period in ischemic stroke.
Initial D-dimer level is a well-known prognostic parameter in patients with acute ischemic stroke (AIS). However, there have been no studies on the clinical significance of follow-up D-dimer levels. In this study, we evaluated the association between initial and follow-up D-dimer levels and early neurological deterioration (END) in patients with AIS. We included consecutive patients with AIS who had a positive initial D-dimer test (> 0.55 mg/L) between March 2021 and November 2022. The follow-up D-dimer test was performed on the 7th day after hospitalization and on the day of discharge if discharged earlier. END was defined as an increase of ≥ 2 in the total NIHSS score, or ≥ 1 in the motor NIHSS score within the first 7 days of admission. As medical conditions closely associated with the initial and follow-up D-dimer levels in AIS patients, we also evaluated the history of cancer, active cancer, and venous thromboembolism (VTE) that occurred during hospitalization together. A total of 246 patients with AIS were evaluated (median age: 87 years, male: 56.5%). In multivariable logistic regression analysis, the initial D-dimer level was closely associated with END after adjusting for confounders (adjusted odds ratio [aOR]: 1.48, 95% CI: 1.06-2.05). The follow-up D-dimer level also showed a close correlation with END (aOR: 1.60, 95% CI: 1.16-2.20). Regarding the analysis of the association between D-dimer levels and underlying cancer or VTE, the initial D-dimer level showed a statistically significant positive relationship only with active cancer (P = 0.024). On the other hand, the follow-up D-dimer level was found to be statistically significantly associated with a history of cancer (P = 0.024), active cancer (P = 0.001), and VTE (P = 0.001). Initial and follow-up D-dimer levels were associated with END in AIS patients. Particularly, the follow-up D-dimer level showed a clear correlation not only with END but also with the underlying cancer or the occurrence of VTE during the acute period.


Mayo Clinic experience with modified Vecchietti procedure for vaginal agenesis: it is easy, safe, and effective.
To evaluate the surgical outcome and the anatomic and sexual function in 10 women with Rokitansky syndrome who underwent the laparoscopic Vecchietti procedure at our center. Retrospective analysis. Data were analyzed on the basis of short-term and long-term surgical outcome and sexual function. All patients underwent clinical follow-up at 1, 2, and 6 months after surgery. In all 10 patients, the procedure produced anatomic and functional success. The laparoscopic Vecchietti technique is safe, simple, and effective for treatment of vaginal agenesis. Results are comparable to those of all European studies, and the procedure should gain more popularity in North America.


Multifunctional ginsenoside Rg3-based liposomes for glioma targeting therapy.
Liposomes have been widely used for targeted drug delivery. However, nonselective distribution, low blood-brain barrier penetration, and the disadvantages of cholesterol greatly limit the application of conventional liposomes in the treatment of brain tumors. In the present study, we aimed to develop a multifunctional ginsenoside Rg3-based liposomal system (Rg3-LPs). Compared to cholesterol liposomes (C-LPs), Rg3-LPs not only significantly improved cellular uptake and penetration across glioma spheroids in vitro, but also remarkably enhanced active glioma targeting and intratumoral diffusion capability in vivo. Paclitaxel-loaded Rg3-LPs (Rg3-PTX-LPs) exhibited a substantially stronger anti-proliferation effect on C6 glioma cells than paclitaxel-loaded C-LPs and re-educated tumor-associated macrophages from the protumor M2 phenotype to the antitumor M1 phenotype in vivo. Rg3-PTX-LPs significantly prolonged median survival time of intracranial C6-bearing mice/rats by activating the immune microenvironment in glioma, facilitating T-cell immune responses with expansion of the CD8+ T-cell population, increasing the M1/M2 ratio, and decreasing regulatory T and myeloid-derived suppressor cells. Together, the results demonstrated that ginsenoside Rg3 is a good alternative for cholesterol in drug delivery liposomes and has a synergistic effect with loaded anticancer drugs. Rg3-PTX-LPs can serve as a multifunctional potential drug for the treatment of glioma.


Canonical Wnt Signaling Modulates the Expression of Pre- and Postsynaptic Components in Different Temporal Patterns.
Wnt ligands play critical roles in neuronal development, synapse formation, synaptic activity, and plasticity. Synaptic plasticity requires molecular remodeling of synapses, implying the expression of key synaptic components. Some studies have linked Wnt signaling activity to changes in synaptic protein levels. However, the presynaptic and postsynaptic gene expression profiles of hippocampal neurons exposed to Wnt proteins have not been studied. Hence, we treated rat cultured hippocampal neurons with recombinant Wnt3a, lithium, and the Wnt inhibitor Dkk-1 for different treatment durations and measured the mRNA and protein levels of pre- and postsynaptic components. The ligand Wnt3a promoted the differential temporal expression of genes encoding presynaptic and postsynaptic proteins. Gene expression of the presynaptic proteins Rim1, piccolo (Pclo), Erc2, Ctbp1 and Rimbp2 increased in a specific temporal pattern. Simultaneously, the mRNA and protein levels of postsynaptic components showed a different temporal expression pattern, e.g., the mRNAs for postsynaptic scaffolding components such as postsynaptic density protein-95 (PSD-95/Dlg4), Homer1 and Shank1 were temporally regulated by both Wnt3a and lithium. On the other hand, the mRNA levels of the gene encoding the protein calcium/calmodulin-dependent protein kinase IV (Camk4), canonically upregulated by Wnt, were increased. Our results suggest that Wnt signaling orchestrates expressional changes in genes encoding presynaptic and postsynaptic components, probably as part of a synaptic plasticity mechanism in neurons.


Multimodal Imaging, OCT En Face, and OCT Angiography of an Anomalous Retinal Artery: Case Report and Review of the Literature.
The purpose is to study for the first time the vascular plexuses and the retinal nerve fiber layer and raphe of a patient with a very uncommon anatomical variation: an anomalous retinal artery supplying the whole macula. We used multimodal imaging, en face spectral-domain optic coherence tomography, and spectral-domain optic coherence tomography angiography. One patient presented in his left eye a very unusual anatomical variation of macular vascularization. A retinal artery deriving from the inferior temporal retinal artery irrigated the whole macula. The formation of the papillomacular bundle and the temporal raphe nerve fiber layer has been attributed to the earlier development of the central retina and to the existence of 2 distinct watershed zones. However, there are very uncommon anatomical variations of the retinal vasculature in which large retinal vessels cross the raphe and could influence the morphology and structure of the nerve fiber layer of the posterior pole. We review the literature on the subject and document for the first time an anomalous artery that irrigates the whole macula, normal thickness and morphology of the nerve fiber layer, and the temporal raphe.


The association of serum neutrophil markers and acute coronary syndrome.
An association exists between chronic infection-induced inflammation, such as periodontitis, and acute coronary syndrome (ACS). We studied the association of serum neutrophil markers, myeloperoxidase (MPO), matrix metalloproteinase (MMP)-8, tissue inhibitor of metalloproteinase (TIMP)-1 concentrations and MMP-8/TIMP-1 ratio, with the risk of recurrent ACS. Radiographic periodontal status was recorded from 141 patients with acute non-Q-wave infarction or unstable angina pectoris, who participated in a double-blind, placebo-controlled study with clarithromycin for 3 months. Serum samples were collected within arrival to the hospital, at 1 week, 3 months and 1 year. Recurrent ACS events were registered during the 1-year follow-up. In the whole population, high serum MPO concentrations at 1 week (fourth quartile versus quartiles 1-3) were associated with the risk of recurrent ACS with a relative risk (RR) of 2.52 (95% CI, 1.277-4.980; P = 0.008). In patients without periodontal disease, high MPO concentration at 1 week and 1 year predicted recurrent ACS with RRs of 3.54 (1.600-7.831; P = 0.002) and 2.87 (1.171-7.038; P = 0.021), respectively. In the placebo group, but not in the clarithromycin group, high serum MMP-8/TIMP-1 ratio at 1 week predicted recurrent ACS with an RR of 3.23 (1.295-8.063; P = 0.012). Our results suggest that high serum neutrophil markers reflect increased risk of recurrent ACS, especially in patients without periodontal disease and not receiving antimicrobial medication.


A model for the thermal inactivation of micro-organisms.
To mathematically model published thermal inactivation data sets using an empirical model based on a double Arrhenius function. A mathematical model, the log R-fat, provided an excellent description of the data sets available: the thermal inactivation of Salmonella anatum at 55 degrees C, Pseudomonas viscosa at 48 degrees C and Streptococcus faecalis at 60 degrees C; Clostridium botulinum spores at various temperatures in the range of 101-121 degrees C (two data sets); thermal inactivation of Salmonella Bedford over the temperature range 50-58 degrees C, water activity range of 0.94-0.99 and a pH range of 4-7; Bacillus stearothermophilus spores from 105 to 121 degrees C and the dry heat sterilization of an indigenous mesophilic soil population over the temperature range of 120-160 degrees C. The log R-fat model, derived from previously published chemical inactivation studies provides as good, if not better, description of thermal inactivation kinetics as other published models. The model does not invoke either of the two hypotheses of inactivation: the mechanistic or vitalistic, although it is closely linked to descriptions of the former. The log R-fat double Arrhenius function provides the investigator with a relatively simple and easy mathematical model to apply to data of thermal inactivation. This model may allow a more accurate description of thermal food processing, especially when the safety of marginal heat processes are concerned.


Management of delayed postoperative hemorrhage after pancreaticoduodenectomy: a meta-analysis.
To determine whether interventional radiology (IR) or laparotomy (LAP) is the best management of delayed postoperative hemorrhage (DPH) after pancreaticoduodenectomy. Data Source We undertook an electronic search of MEDLINE and selected for analysis only original articles published between January 1, 1990, and December 31, 2007. Two of us independently selected studies reporting on clinical presentation and incidence of postoperative DPH and the following outcomes: complete hemostasis, morbidity, and mortality. Two of us independently performed data extraction. Data were entered and analyzed by means of dedicated software from The Cochrane Collaboration. A random-effects meta-analytical technique was used for analysis. One hundred sixty-three cases of DPH after pancreaticoduodenectomy were identified from the literature. The incidence of DPH after pancreaticoduodenectomy was 3.9%. Seventy-seven patients (47.2%) underwent LAP; 73 (44.8%), IR; and 13 (8%), conservative treatment. On meta-analysis comparing LAP vs IR for DPH, no significant difference was found between the 2 treatment options for complete hemostasis (73% vs 76%; P = .23), mortality (43% vs 20%; P = .14), or morbidity (77% vs 35%; P = .06). This meta-analysis, although based on data from small case series, is unable to demonstrate any significant difference between LAP and IR in the management of DPH after pancreaticoduodenectomy. The management of this life-threatening complication is difficult, and the appropriate treatment pathway ultimately will be decided by the clinical status of the patient and the institution preference.


Genetics of human C4 polymorphism: detection and segregation of rare and duplicated haplotypes.
Applying a combined technology for the detection of allotypic variation of the fourth component of human complement (C4), including immunofixation with anti-C4 and C4-dependent lysis after agarose electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of C4 to separate the C4A and B alpha-chains, and the determination of Rodgers (Rg) and Chido (Ch) determinants of C4 in serum and at the blotted C4 alpha-chains, we detected rare human C4 allotypes and studied the genetic linkage. Partial inhibitors (p.i.) of anti-Rg and anti-Ch sera were found; the C4A51 allotype characterized as Rg p.i. and the C4A1 and C4B51 allotypes as Ch p.i. were genetically inherited. The C4A1 allotype has a unique Rg- Ch+ C4A alpha-chain. Duplicated C4A loci, A*3, A*2, and A*5, A*2 were both associated with a C4BQO and the HLA haplotype A3-Cw4-Bw35-DR1. These additions to the already known extensive C4 polymorphism may help to sort out their significance for the biological functions of human C4.


Perforation of a central venous catheter into the pelvis of the kidney: a rare complication.
Most adverse events related to central venous catheter (CVC) use in severely ill neonates who require total parenteral nutrition (TPN) are caused by malposition of the tip of the catheter, followed by complications such as infection, sepsis, thrombosis or, rarely, perforation of the vessel. Malpositioned tip of a CVC into the renal vein followed by thrombosis is well known. We report two cases of perforation of a CVC into the renal pelvis. To prevent this complication, the value of intra- or postoperative use of fluoroscopy with injection of contrast medium through the catheter is discussed.


Further studies on a native Australian eucalypt forest invaded by exotic pines.
Pinus radiata is spreading from plantations into some types of native eucalypt forest. A broad transect through one heavily invaded site was mapped in 1974 and re-examined after intervals of time. In 1977 the adjacent pine plantation was clear-felled for commercial purposes. By 1981 it was clear that the pines that had already invaded the eucalypt forest were contributing their own progeny to the mixed stand. However, the rate of recruitment is slow due to high seedling mortality. Comparative measurements have shown that during the period 1974-1981 the growth rate of pines established among the eucalypts was approximately ten times greater than that of the native trees.


A saliency map in primary visual cortex.
I propose that pre-attentive computational mechanisms in primary visual cortex create a saliency map. This map awards higher responses to more salient image locations; these responses are those of conventional V1 cells tuned to input features, such as orientation and color. Hence no separate feature maps, or any subsequent combination of them, is needed to create a saliency map. I use a model to show that this saliency map accounts for the way that the relative difficulties of visual search tasks depend on the features and spatial configurations of targets and distractors. This proposal links psychophysical behavior to V1 physiology and anatomy, and thereby makes testable predictions.


Limonoids from Swietenia macrophylla and their antitumor activities in A375 human malignant melanoma cells.
Swietelinins A - C (1-3) and swieteliacates F - R (4-16), sixteen new limonoids and 18 known limonoids (17-34) were isolated from Swietenia macrophylla. The absolute configurations of these compounds were defined by using a combination of electronic circular dichroism data analysis and single-crystal X-ray diffraction data. Swieteliacate J (10) is the first limonoid possessing an unusual 8β, 9β-epoxy ring system. All of the compounds were tested for cytotoxicity against four human tumor cell lines (SMMC-7721, SW620, A549, and A375). Compounds 10, 11, and 19 exhibited selectively moderate cytotoxicity against four tumor cell lines, especially 19 exhibited significant cytotoxic effects against A375 with IC50 an value of 9.8 μM and was more active than the positive control, dacarbazine with an IC50 value of 22.4 μM. Compound 19 effectively induced apoptosis of A375, which was associated with G2/M-phase cell cycle arrest. Flow cytometric analysis showed that the treatment by 19 significantly induced A375 cell apoptosis in a dose-dependent manner.


Low incidence of null alleles of the fourth component of complement (C4) in elderly people.
Allotype frequencies of three complement proteins (C2, factor B, and C4) encoded on the sixth chromosome were tested in 150 "young" (under 53 yrs) and 131 "old" (over 62 yrs) healthy Hungarian individuals. In women, we found significant differences in BF*F allotype frequencies between the two age groups. A marked decrease of null alleles for C4 was observed especially in men. In the "old" age group, the total frequency of the non-expressed, "null" (Q0) alleles of the C4 protein dropped to almost one third of the "young" ones. The decrease was even larger (a 5.5-fold decrease in the incidence) among men. The relative risk of those having C4B*Q0 allele not to survive a "critical age period" of 53-62 yrs for males and 58-62 yrs for females was calculated to be 12.4 and 2.85, respectively. Our data substantiate the view that the expected life time is genetically determined and it can be forecast partly by the analysis of the antigens encoded on the sixth chromosome.


On the biomechanical function of the liver capsule.
We have carried out histomechanical studies on the human bovine liver caps-le under tension loading. Nonlinear force-time curve under extension with constant speed, initial upper and lower summit decrease of the dynamic relaxation curve, amplitude diminution phenomenon of the dynamic relaxation in the lower nonlinear part of the extension-time curve, dynamic (cyclic) force recovery.


Determination of tryptophan and histidine by adsorptive cathodic stripping voltammetry using H-point standard addition method.
A sequential method is proposed for the determination of tryptophane and histidine by adsorptive cathodic stripping voltammetry using standard addition and H-point standard addition method (HPSAM). The complexes of copper(II) with the amino acids were accumulated onto the surface of a hanging mercury drop electrode for 60s. Then the preconcentrated complexes were reduced by square wave voltammetry and the peak currents were measured. The effect of various parameters such as pH, concentration of copper, accumulation potential, accumulation time and scan rate on the sensitivity were studied by one-at-a time and artificial neural network. Under the optimized conditions, the peak currents at about +0.05 to -0.30 V is proportional to the concentration of tryptophan and histidine over the concentration ranges of 5-220 and 100-1200 nM, respectively. Optimization of the parameters by one-at-a time showed that at accumulation potential of 0.10 V (versus Ag/AgCl reference electrode) the peak current is proportional only to the concentration of tryptophan and histidine does not have any contribution to the current. The optimization results by artificial neural network showed that at accumulation potential of -0.06 V (versus Ag/AgCl) the peak current is proportional to the both concentrations of tryptophan and histidine. Therefore, the method of H-point standard addition has been used for resolving overlap voltamograms for determination of histidine in the present of tryptophane. The method was successfully applied to the determination of tryptophan and histidine in synthetic and real samples.


Value of liver scintiscan in staging of testicular neoplasm.
A nineteen-year retrospective study of the usefulness of liver scanning in the staging evaluation of germinal cell testicular neoplasms was undertaken at the National Naval Medical Center. Of 94 patients, 90 (96 per cent) demonstrated accurate correlation between liver scan and histopathologic diagnosis.


Analysis of Neutrophil Bactericidal Activity.
This chapter describes three methods for measuring the bactericidal activity of neutrophils. All utilize colony counting techniques to quantify viable bacteria. A simple "one-step" protocol provides a composite measure of phagocytosis and killing, while a "two-step" protocol that separates extracellular and intracellular bacteria allows calculation of rate constants for both of these processes. We also present a method for selectively monitoring the long-term survival of bacteria within the phagosome. This may have application in identifying resistant strains and searching for compounds that sensitize pathogens to destruction.


One registration multi-atlas-based pseudo-CT generation for attenuation correction in PET/MRI.
The outcome of a detailed assessment of various strategies for atlas-based whole-body bone segmentation from magnetic resonance imaging (MRI) was exploited to select the optimal parameters and setting, with the aim of proposing a novel one-registration multi-atlas (ORMA) pseudo-CT generation approach. The proposed approach consists of only one online registration between the target and reference images, regardless of the number of atlas images (N), while for the remaining atlas images, the pre-computed transformation matrices to the reference image are used to align them to the target image. The performance characteristics of the proposed method were evaluated and compared with conventional atlas-based attenuation map generation strategies (direct registration of the entire atlas images followed by voxel-wise weighting (VWW) and arithmetic averaging atlas fusion). To this end, four different positron emission tomography (PET) attenuation maps were generated via arithmetic averaging and VWW scheme using both direct registration and ORMA approaches as well as the 3-class attenuation map obtained from the Philips Ingenuity TF PET/MRI scanner commonly used in the clinical setting. The evaluation was performed based on the accuracy of extracted whole-body bones by the different attenuation maps and by quantitative analysis of resulting PET images compared to CT-based attenuation-corrected PET images serving as reference. The comparison of validation metrics regarding the accuracy of extracted bone using the different techniques demonstrated the superiority of the VWW atlas fusion algorithm achieving a Dice similarity measure of 0.82 ± 0.04 compared to arithmetic averaging atlas fusion (0.60 ± 0.02), which uses conventional direct registration. Application of the ORMA approach modestly compromised the accuracy, yielding a Dice similarity measure of 0.76 ± 0.05 for ORMA-VWW and 0.55 ± 0.03 for ORMA-averaging. The results of quantitative PET analysis followed the same trend with less significant differences in terms of SUV bias, whereas massive improvements were observed compared to PET images corrected for attenuation using the 3-class attenuation map. The maximum absolute bias achieved by VWW and VWW-ORMA methods was 06.4 ± 5.5 in the lung and 07.9 ± 4.8 in the bone, respectively. The proposed algorithm is capable of generating decent attenuation maps. The quantitative analysis revealed a good correlation between PET images corrected for attenuation using the proposed pseudo-CT generation approach and the corresponding CT images. The computational time is reduced by a factor of 1/N at the expense of a modest decrease in quantitative accuracy, thus allowing us to achieve a reasonable compromise between computing time and quantitative performance.


Efficacy of stereotactic radiosurgery as single or combined therapy for brain metastasis: A systematic review and meta-analysis.
To determine the efficacy of stereotactic radiosurgery (SRS) in treating patients with brain metastases (BMs), a network meta-analysis (NMA) of randomized controlled trials (RCTs) and a direct comparison of cohort studies were performed. Relevant literature regarding the effectiveness of SRS alone and in combination with whole-brain radiotherapy (WBRT) and surgery was retrieved using systematic database searches up to April 2019. The patterns of overall survival (OS), one-year OS, progression-free survival (PFS), one-year local brain control (LBC), one-year distant brain control (DBC), neurological death (ND), and complication rate were analyzed. A total of 18 RCTs and 37 cohorts were included in the meta-analysis. Our data revealed that SRS carried a better OS than SRS+WBRT (p = 0.048) and WBRT (p = 0.041). Also, SRS+WBRT demonstrated a significantly improved PFS, LBC, and DBC compared to WBRT alone and SRS alone. Finally, SRS achieved the same LBC as high as surgery, but intracranial relapse occurred considerably more frequently in the absence of WBRT. However, there were not any significant differences in ND and toxicities between SRS and other groups. Therefore, SRS alone may be a better alternative since increased patient survival may outweigh the increased risk of brain tumor recurrence associated with it.


Two interleukin (IL)-15 homologues in fish from two distinct origins.
Here, we report two distinct genes in teleosts that are homologous to interleukin (IL)-15. The two genes, isolated from fugu (Takifugu rubripes), resemble to mammalian IL-15 but differ from IL-2 and IL-21 in their amino acid sequences, the possessing of an extraordinary long signal peptide and more widespread tissue localization. In addition, multiple out-of-frame AUG codons, the negative translational regulators of mammalian IL-15 genes were also detected in the 5'-UTR of the two genes. Fugu IL-15 homologues also contain four conserved cysteines allowing the formation of two disulfide bridges along with four predicted alpha-helices. Genomic analysis showed that one of the fugu IL-15 homologues possessed six coding exons and exhibited a similar exon-intron organization and synteny structure to that of mammalian and chicken IL-15 genes. Conversely, the other fugu IL-15 homologue possesses four exons and exhibits a different synteny structure with that of IL-15, suggesting that the two genes were derived from two different origins. Moreover, the two genes also differ from each other in tissue localizations and in their expression in response to mitogens. The existence of these two IL-15 homologues in telesots was further supported by their characterization in zebrafish Danio rerio, and the green-spotted pufferfish Tetraodon nigroviridis. The discovery of two distinct IL-15 homologues in fish will assist investigations into the evolution of these genes and their relative contribution to the fish immune system.


Beneficial role of pre- and post-ozonation in a low rate biofiltration-ultrafiltration process treating reclaimed water.
Previous studies have shown that the combination of biological and ozone oxidation processes can achieve a greater performance in treating natural surface water than each process individually. In this work, we designed and tested an ozonation-gravity-driven up-flow slow rate (0.01 m/h) biofiltration-ozonation (O3-GUSB-O3) process for the pre-treatment of reclaimed water prior to ultrafiltration (UF), with the aim of producing high quality drinking water and a significantly reduced degree of UF fouling. Results showed that O3 coupled with GUSB can effectively remove aromatic compounds (∼ 84.8%), dissolved organic carbon (DOC, ∼ 83.4%), and biopolymers in surface water. In addition, post-ozonation greatly contributed to the reduction of the UF membrane fouling (∼ 6 times greater flux). With regard to the disinfection by-product formation potential (DBPFP) of the final treated water, both trihalomethane formation potential (THMFP) and haloacetic acid formation potential (HAAFP) were greatly reduced (86.4% and 84.8% for THMs and HAAs, respectively). The relationship between DBPFP and various spectral indexes revealed that aromatic compounds and amino acids were more likely to generate DBPs during the disinfection stage. Among these, humic substances were more likely to generate THMs, while low molecular weight carboxylate and carbonyl organic compounds were associated with the generation of HAAs. Moreover, the dosage of O3 during the post-ozonation stage was found to influence directly the generation of DBPs. Overall, this study has conducted a detailed evaluation of a novel multi-ozone biofilter UF process for treating surface water, and the results provide a valuable basis for subsequent studies at larger scale to demonstrate the potential of the treatment process for practical applications.


Human immunodeficiency virus infection in pregnancy: epidemiology and prevention of vertical transmission.
The preponderance of AIDS cases in women during their reproductive years demands attention to primary prevention, early recognition, and appropriate interventions. Increases in the epidemic among injection drug users and heterosexually exposed persons have resulted in an increased impact of HIV/AIDS among women and children. It is clear that vertical transmission of HIV can occur early in pregnancy, during labor and delivery, or in the postpartum period. For these reasons, prevention strategies must be comprehensive and include primary prevention and appropriate intervention during pregnancy and in the peripartum and postpartum area. Stringent infection control techniques are imperative during delivery and in the postpartum period, even in the absence of clear data supporting this intervention. Recent data suggest that the cost of treating one adult patient with AIDS is approximately $100,000, making prevention and intervention imperative.


Role of the vitamin K epoxide reductase complex subunit 1 (VKORC1) -1639G>A gene polymorphism in patients with retinal vein occlusion.
Retinal vein occlusion (RVO) is a major vision-threatening disease. Vitamin K epoxide reductase recycles reduced vitamin K, which is essential for the gamma carboxylation of clotting factors II, VII, IX, X and proteins C and S. Recently, the vitamin K epoxide reductase complex subunit 1 (VKORC1) -1639G>A (rs9923231) polymorphism has been reported as a novel risk factor for RVO in a Turkish population. The present study was set to confirm or to refute this association in a larger cohort of patients with RVO. The present case-control study comprised 285 patients with central RVO, 401 patients with branch RVO and 333 control subjects. Genotypes of the VKORC1 -1639G>A polymorphism were determined by 5' exonuclease assay (TaqMan). No significant differences in either genotype distributions or allele frequencies of the vitamin K epoxide reductase complex subunit 1 -1639G>A polymorphism were found between patients and control subjects (p > 0.05). In a logistic regression analysis neither branch nor central RVO was predicted by the vitamin K epoxide reductase complex subunit 1 -1639G>A genotypes, but by arterial hypertension, ever-smoking status and in case of central RVO additionally by diabetes mellitus. Our data suggest that the vitamin K epoxide reductase complex subunit 1 -1639G>A gene polymorphism is unlikely a major risk factor for patients with either central or branch RVO.


Plasma metabolites and food intake reduction following heparinoid injection in rats.
Behavioral and physiological consequences of heparinoid injections, which increase plasma levels of nonesterified fatty acids (NEFA) by releasing lipoproteinlipase into the circulation, were studied in rats, because, according to the lipostatic hypothesis of food intake control, changes in plasma metabolite levels should influence food intake. Subcutaneously injected 55 mg/kg body wt. of the heparinoid Na-pentosanpolysulfate (PPS) significantly reduced feeding in rats with ad lib access to either a high carbohydrate (HC) or a high fat (HF) diet. However, the reduction of food intake in HF-rats exceeded that in HC-rats. Moreover, the food intake reduction following PPS-injections was associated with greater increases in plasma levels of NEFA and free glycerol in HF-rats, whereas PPS increased plasma levels of D-(-)-3-hydroxybutyrate only in HF-rats, and did not alter blood glucose concentrations. Although PPS inhibited blood coagulation, it did not affect the hematocrit. Furthermore, PPS-injections that reduced feeding, failed to produce a conditioned taste aversion in a two bottle preference test. The data therefore strongly suggest that elevated levels of plasma NEFA and/or free plasma glycerol decrease food intake in rats. These metabolites might contribute to a lipostatic mechanism of feeding control.


The use of selection in recovery of transgenic targets for mutation analysis.
Transgenic animal mutagenesis assays using lambda shuttle vectors have recently been described for isolation and characterization of spontaneous and chemical induced DNA mutations. Extensive information on lambda and E. coli genetics provides a wealth of techniques to allow selection of mutant target genes. Here we describe the modification of an E. coli host which permits two methods for the direct selection of mutant genes. These methods reduce the number of plates needed to be screened for a comparable amount of frequency data by 20-100-fold and thus provide a significant savings of the materials and time required for the screening of mutations. In addition, mutants selected by these approaches described here may alter or broaden the spectrum of mutations that are recoverable. Ultimately, a combination of selective and nonselective techniques may prove valuable for the analysis of mutations produced in vivo in transgenic animals.


Temporal variability of chlorinated volatile organic compound vapor concentrations in a residential sewer and land drain system overlying a dilute groundwater plume.
Some subsurface sewer and land drain networks will facilitate the migration of chlorinated volatile organic compounds (CVOCs) from dissolved contaminant groundwater plumes to indoor air. As this vapor intrusion (VI) pathway has only recently been documented, guidance for evaluating it, including recommendations for timing, frequency, duration and location for vapor sampling in subsurface piping networks is non-existent. To address this gap, a three-year investigation of CVOC concentrations from land drains, storm drains, and sanitary sewers was undertaken in a neighborhood overlying a large-scale dissolved chlorinated VOC (CVOC) groundwater plume. Vapor sampling included the collection of grab (time-discrete) samples from up to 277 manholes, hourly grab sampling from three manhole locations, and 24-h duration collection during week-long sampling from 13 land drain and sewer manholes. The spatial distribution of vapor and water concentrations and the temporal variations in the vapor values observed in this study suggest that week-long vapor sampling conducted at different times of the year and with samples collected at manhole locations overlying and outside a dissolved plume might be needed to ensure robust VI pathway assessment at other sites. These findings are expected to be of relevance to regulatory agencies involved in the development of current or future VI pathway assessment guidance.


Anterior urethral valves: an uncommon cause of obstructive uropathy in children.
Anterior urethral valves (AUV) are rare entities generally described in case reports. They are an uncommon cause of lower urinary tract obstruction in children and can be difficult to diagnose. In the present study, we present our experience in four children with AUV along with a literature review. We retrospectively identified four children with AUV presented between 1998 and 2005 at age 4-9 years. Hematuria, urinary tract infection and weak voiding stream were the most common symptoms. Voiding cystourethrography (VCUG) confirmed the diagnosis of AUV. On cystourethroscopy, cusp-like valves in the anterior urethra were seen in all children. Transurethral endoscopic resection of the valves was carried out in three children using a pediatric resectoscope. In one child with a massive anterior urethral diverticulum, open resection of the valve, diverticulectomy and urethroplasty were performed. All patients were cured, none had complications as a result of surgery, and all reported a normal urinary stream at follow-up. Children with poor stream and recurrent infections should be evaluated carefully and anterior urethral valves should be considered in differential diagnosis of obstructive lesions.


Essential role of excessive tryptophan and its neurometabolites in fatigue.
Serotonin, a neurotransmitter synthesized from tryptophan, has been proposed to play a key role in central fatigue. In this study, we examined whether tryptophan itself and/or its two metabolites, kyneurenic acid (KYNA) and quinolinic acid (QUIN), are involved in central fatigue. Experiments were conducted using Sprague-Dawley rats (SDR) and Nagase analbuminemic rats (NAR). Central fatigue was assessed by treadmill running and a Morris water maze test. Microdialysis was used to collect samples for measurement of extracellular concentration of tryptophan, serotonin and 5-hydroxyindoleacetic acid (5-HIAA) and to infuse test agents. To examine the kinetics of release, synaptosomes in the striatum were prepared in vitro to measure intra- and extrasynaptosomal concentration of tryptophan, serotonin and 5-HIAA. The concentration of tryptophan secreted into the extracellular space of the striatum was higher during fatigue only, and quickly returned to basal levels with recovery from fatigue. Running time to exhaustion was reduced by activation of tryptophan receptors. Time to exhaustion was shorter in NAR, which maintain a higher extracellular level of striatum tryptophan than SDR. Impaired memory performance in a water maze task after tryptophan treatment was attributable to high levels of KYNA and QUIN in the hippocampus acting synergistically on N-methyl-D-aspartic acid receptors. When branched-chain amino acids were administered, tryptophan transport to the extracellular space of the striatum was drastically inhibited. Our findings demonstrate that the increase in fatigue which occurs because of excessively elevated brain tryptophan can be further amplified by the use of synthetic KYNA and QUIN.


[Use of propiono-acidophilus milk in the complex treatment of intestinal dysbacteriosis in infants with staphylococcal infections and sepsis].
Antistaphylococcal properties of the new lactic acid mixture propionic acidophilic milk (PAM) against the microbe isolated from feces of children suffering from staphylococcal sepsis were comparatively studied by the dilution and diffusion methods. PAM was found to have more pronounced antimicrobial properties against pathogenic staphylococcus than acidophilic milk and kefir. It is recommended to include PAM into the diet of children with intestinal dysbacteriosis.


Removal of 14-C-palmitate during ventriculocisternal perfusion in conscious dogs in insulin-induced hypoglycemia.
A detailed description of a method is presented allowing continous ventriculocisternal perfusion of metabolites in the conscious dog. Using this preparation, the loss of the infused albuminbound 14-C-aplmitate, was studied in 14 conscious dogs during ventriculocisternal perfusion with artificial cerebrospinal fluid (CSF). Forty-five percent of the infused 14-C-palmitate was recovered from the cisternal effluent under equilibrium conditions after 60 min of perfusion. The effect of the injection of 2.0 U insulin/kg was also investigated in seven dogs. Plasma glucose concentration decreased to 40--50 mg% and the amount of 14-C-palmitate recovered was significantly higher one hour after insulin compared to the controls receiving saline injections. No significant changes in cerebral arteriovenous differences of glucose or oxygen or in venous 14-C-palmitate concentration were observed during the same time. It is concluded that the combined use of ventriculocisternal perfusion and the analysis of cerebral arteriovenous differences are useful in studying brain metabolism in the consciuos dog.


Re-evaluating high-frequency oscillation for ARDS: Would a targeted approach be successful?
High-frequency oscillatory ventilation (HFOV) seems ideal for lung protection in acute respiratory distress syndrome, but randomized trials have not shown a mortality reduction. The initial oxygenation response to HFOV appears to be associated with survival. Here, we discuss the mechanisms of oxygenation response to increases in airway pressure and the interpretation of the oxygenation response observed in the present study.


Documentation of pediatric drug safety in manufacturers' product monographs: a cross-sectional evaluation of the canadian compendium of pharmaceuticals and specialities.
To describe the provision of pediatric drug safety information in a national formulary of manufacturers' drug product monographs. We performed a cross-sectional evaluation of comprehensive product monographs contained in the 2005 Canadian Compendium of Pharmaceuticals and Specialities (CPS). We abstracted data describing indications for prescription, statements about pediatric safety, available preparations, and provision of dosing guidelines. For each monograph we classified pediatric safety data as either present, present but limited or absent. We then described the pediatric safety data in CPS monographs for drugs listed in the published formulary of the Hospital for Sick Children, Toronto, Ontario, Canada. A total of 2232 product monographs were screened; 684 were excluded and 1548 (66%) were further analyzed. 1462 (94%) had indications that did not exclude children. Pediatric safety information was present in 592 (38%), present but limited in 148 (10%), and absent in 808 (52%) drug monographs. Safety statements were absent in 224 (14%) drug monographs that provided both dosing guidelines and formulations suitable for administration to children, and in 214 (52%) of 411 drugs in the pediatric hospital formulary. We evaluated a widely available national source of pediatric prescribing information. Safety data for children was not mentioned in more than half of the product monographs. Moreover, the provision of safety data was discordant with indications for prescription, the availability of pediatric formulations, and dosing guidelines within the monographs, and with inclusion in a pediatric hospital formulary. Our study suggests that the presentation of pediatric safety data in drug product monographs can be improved to better inform prescribing and to optimize pharmacotherapy in children.


The incidence of red cell alloantibodies underlying panreactive warm autoantibodies.
A recognized hazard of administering blood transfusions to patients with panreactive warm autoantibodies is that alloantibodies may be masked. Studies have shown the incidence of underlying alloantibodies to be 30 to 40 percent. Adsorption procedures can be used to remove autoantibodies and allow detection and identification of underlying alloantibodies. This study contains data from 126 patients referred to the Red Cell Immunohaematology laboratory at the National Blood Service, Newcastle upon Tyne, United Kingdom. These patients were from the northeast of England, a population for which data have not previously been reported. Samples identified as containing panreactive warm autoantibodies were subjected to adsorption procedures (95 by alloadsorption and 31 by autoadsorption). Absorbed sera were then tested to identify underlying alloantibodies. Of 126 samples, 39 (31%) contained a total of 61 RBC alloantibodies; 15 (12%) contained 2 or more antibody specificities; and 14 (11%) contained alloantibodies not found within the Rh or Kell blood group systems. Antibodies identified included the following specificities: E (19), D (9), c (7), C (6), S (5), Fy(a) (3), Jk(a) (2), Jkb (2), K (2),Kp(a) (2), Fy(b),C(w),N, and f (ce). This study reinforces the value of adsorption studies, whether using autologous or allogeneic RBCs, when panreactive warm autoantibodies are present. In addition, this study confirms that it is not appropriate in these cases simply to issue blood which is "least incompatible" or Rh phenotype- and K antigen-matched.


Integrated imaging of hepatic tumors in childhood. Part II: Benign lesions (congenital, reparative, and inflammatory).
The authors have encountered benign liver masses as frequently as malignant lesions in children with hepatomegaly. Lesions studied included abscesses, cavernous hemangioma/hemangioendothelioma, adenoma of glycogen storage disease, choledochal cysts, focal nodular hyperplasia, cystic hepatoblastoma, and hamartoma. An integrated imaging protocol involving ultrasound, computed tomography, and scintigraphy proved to be more helpful than any one modality in establishing the benign or malignant nature of a hepatic neoplasm and the type of tumor, which is of particular importance when surgical exploration and/or biopsy is contraindicated.


Real-time control of purified product collection during chromatography of recombinant human insulin-like growth factor-I using an on-line assay.
During preparative reversed-phase chromatography of recombinant human insulin-like growth factor-I (IGF), the separation of IGF from IGF aggregates cannot be determined using UV absorbance. An on-line reversed-phase chromatographic assay was developed that provides a quantitative measurement of IGF and IGF aggregates every 4 min, allowing real-time control of purified IGF collection. Process control using the on-line assay is a reliable and accurate method to collect purified IGF.


Internal aeration and respiration of submerged tomato hypocotyls are enhanced by ethylene-mediated aerenchyma formation and hypertrophy.
With the impending threat that climate change is imposing on all terrestrial ecosystems, the ability of plants to adjust to changing environments is, more than ever, a very desirable trait. Tomato (Solanum lycopersicum L.) plants display a number of responses that allow them to survive under different abiotic stresses such as flooding. We focused on understanding the mechanism that facilitates oxygen diffusion to submerged tissues and the impact it has on sustaining respiration levels. We observed that, as flooding stress progresses, stems increase their diameter and internal porosity. Ethylene triggers stem hypertrophy by inducing cell wall loosening genes, and aerenchyma formation seems to involve programmed cell death mediated by hydrogen peroxide. We finally assessed whether these changes in stem morphology and anatomy are indeed effective to restore oxygen levels in submerged organs. We found that aerenchyma formation and hypertrophy not only increase oxygen diffusion toward the base of the plant, but also result in an augmented respiration rate. We consider that this response is crucial to maintain adventitious root development under such conditions and, therefore, making it possible for the plant to survive when the original roots die.


Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings.
Human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) have become an essential tool to study arrhythmia mechanisms. Much of the foundational work on these cells, as well as the computational models built from the resultant data, has overlooked the contribution of seal-leak current on the immature and heterogeneous phenotype that has come to define these cells. The aim of this study is to understand the effect of seal-leak current on recordings of action potential (AP) morphology. Action potentials were recorded in human iPSC-CMs using patch clamp and simulated using previously published mathematical models. Our in silico and in vitro studies demonstrate how seal-leak current depolarizes APs, substantially affecting their morphology, even with seal resistances (Rseal) above 1 GΩ. We show that compensation of this leak current is difficult due to challenges with obtaining accurate measures of Rseal during an experiment. Using simulation, we show that Rseal measures (i) change during an experiment, invalidating the use of pre-rupture values, and (ii) are polluted by the presence of transmembrane currents at every voltage. Finally, we posit that the background sodium current in baseline iPSC-CM models imitates the effects of seal-leak current and is increased to a level that masks the effects of seal-leak current on iPSC-CMs. Based on these findings, we make recommendations to improve iPSC-CM AP data acquisition, interpretation, and model-building. Taking these recommendations into account will improve our understanding of iPSC-CM physiology and the descriptive ability of models built from such data.


Creatine phosphokinase activity in dysgenic (mdg/mdg) mouse muscle.
The specific activity of creatine phosphokinase (CPK) was measured in the muscle of mdg/mdg and control embryos of 14-18 days' gestation. CPK specific activity values were similar in mutant and normal embryos at the earliest stages examined (14-15 days). However, after 15 1/2 days, the specific activity of the enzyme in the mdg/mdg embryos was approximately 50% lower than in the controls. The dysgenic and normal muscle extracts exhibited comparable stability after storage at -85 C. CPK activity levels in the muscle of adult heterozygotes (+/mdg) and wild-type (+/+) controls were found to be statistically identical. The findings suggest that the mdg mutation does not have a primary or direct effect on CPK activity.


Simulating porcelain firing effect on the structure, corrosion and mechanical properties of Co-Cr-Mo dental alloy fabricated by soft milling.
This study aims at evaluating the effect of simulating porcelain firing on the microstructure, corrosion behavior and mechanical properties of a Co-Cr-Mo alloy fabricated by Metal Soft Milling (MSM). Two groups of Co-28Cr-5Mo specimens (25 × 20 × 3 mm) were prepared by MSM: The as-sintered (AS) specimens and the post-fired (PF) specimens that were subjected to 5 simulating porcelain firing cycles without applying the ceramic mass onto their surface. Phase identification by X-ray Diffraction (XRD), microstructure examination by optical microscopy and Scanning Electron Microscopy combined with Energy-Dispersive X-ray Spectroscopy (SEM/EDX), corrosion testing by cyclic polarization and chronoamperometry in simulated body fluid (SBF), the latter test accompanied by Cr3+ and Cr6+ detection in the electrolyte through the 1.5-diphenylcarbazide (DPC) method and UV/visible spectrophotometry, and mechanical testing by micro-/nano-indentation were conducted to evaluate the effect of the post-firing cycles on the properties of Co-Cr-Mo. The results were statistically analyzed by the t test (p < 0.05: statistically significant). All specimens had a mixed γ-fcc and ε-hcp cobalt-based microstructure with a dispersion of pores filled with SiO2 and a fine M23C6 intergranular presence. PF led to an increase in the ε-Co content and slight grain coarsening. Both AS and PF alloys showed high resistance to general and localized corrosion, whereas neither Cr6+ nor Cr3+ were detected during the passivity-breakdown stage. PF improved the mechanical properties of the AS-alloy, especially the indentation modulus and true hardness (statistically significant differences: p = 0.0009 and 0.006, respectively). MSM and MSM/simulating-porcelain firing have been proven trustworthy fabrication methods of Co-Cr-Mo substrates for metal-ceramic prostheses. Moreover, the post-firing cycles improve the mechanical behavior of Co-Cr-Mo, which is vital under the dynamically changing loads in the oral cavity, whereas they do not degrade the corrosion performance.


Immunocytochemical identification of serotonin axonal contacts on characterized neurons in laminae I and II of the cat dorsal horn.
Serotonergic pathways from brainstem to spinal cord play an important role in the modulation of pain perception. To establish where that modulation occurs, we examined serotonin-immunoreactive axonal contacts on individually characterized laminae I and II dorsal horn neurons intracellularly filled with horseradish peroxidase. We found serotonin axonal contacts on marginal, stalked, and islet cells. Contacts preferentially occurred on dendritic shafts rather than on spines. Marginal and stalked cells received the heaviest innervation.


Proliferative index of human luteinized granulosa cells varies as a function of ovarian reserve.
We examined whether the proliferative index of luteinized granulosa cells, as determined by flow cytometry, varied as a function of a woman's ovarian reserve, as reserve, as reflected by follicular-phase day 3 serum follicle-stimulating hormone. This prospective cohort study consisted of 19 women of similar chronologic age preparing for in vitro fertilization-embryo who met specific day 3 serum follicle-stimulating hormone criteria. The "low follicle-stimulating hormone" group consisted of 11 women with day 3 serum follicle-stimulating hormone levels < or = 6 IU/L. The "high follicle-stimulating hormone" group consisted of eight women with day 3 serum follicle-stimulating hormone levels > or = 18 IU/L. A total of 56 preovulatory follicles containing > or = 10(4) luteinized granulosa cells were examined by flow cytometry. The low follicle-stimulating hormone group was compared with the high follicle-stimulating hormone group to examine proliferative index as a function of serum day 3 follicle-stimulating hormone levels. The low follicle-stimulating hormone group had a greater proliferative index (11.1% +/- 0.4%) than did the high follicle-stimulating hormone group (8.3% +/- 0.6%), p < 0.001). This study demonstrates that in spite of the same chronologic age, luteinized granulosa cells from preovulatory follicles of women with day 3 serum follicle-stimulating hormone levels > or = 18 IU/L have a 25% decreased proliferative index compared with luteinized granulosa cells from women with day 3 serum follicle-stimulating hormone levels < or = 6. This suggests that granulosa cell proliferation is influenced by ovarian reserve and may explain in part the more favorable response to ovulation induction protocols that younger women demonstrate compared with women of more advanced reproductive age.


SLAM family receptors control pro-survival effectors in germinal center B cells to promote humoral immunity.
Expression of the signaling lymphocytic activation molecule (SLAM)-associated protein (SAP) is critical for the germinal center (GC) reaction and T cell-dependent antibody production. However, when SAP is expressed normally, the role of the associated SLAM family receptors (SFRs) in these processes is nebulous. Herein, we established that in the presence of SAP, SFRs suppressed the expansion of the GC reaction but facilitated the generation of antigen-specific B cells and antibodies. SFRs favored the generation of antigen-reactive B cells and antibodies by boosting expression of pro-survival effectors, such as the B cell antigen receptor (BCR) and Bcl-2, in activated GC B cells. The effects of SFRs on the GC reaction and T cell-dependent antibody production necessitated expression of multiple SFRs, both in T cells and in B cells. Hence, while in the presence of SAP, SFRs inhibit the GC reaction, they are critical for the induction of T cell-mediated humoral immunity by enhancing expression of pro-survival effectors in GC B cells.


Element-tagged immunoassay with inductively coupled plasma mass spectrometry for multianalyte detection.
The multianalyte immunoassay approach is currently attracting increasing attention due to its high sample throughput, short assay time, low sample consumption and reduced overall cost per assay. This paper reviews progress in the field of multianalyte immunoassays using inductively coupled plasma mass spectrometry, as well as applications of this approach in different fields. Examples related to the combination of protein microarray technology with the multitag approach of the immunoassay ICP-MS method and to the use of ICP-MS in the field of imaging are described. A novel strategy that involves tagging antibodies for ICP-MS detection in sensitive multitag bioassays is also presented. Finally, the outlook for this promising technique is discussed.


Formation of functionalized nanoclusters by solvent evaporation and their effect on the physicochemical properties of dental composite resins.
The aim of this work was to study the effect of silica nanoclusters (SiNC), obtained by a solvent evaporation method and functionalized by 3-methacryloxypropyltrimethoxysilane (MPS) and MPS+octyltrimethoxysilane (OTMS) (50/50wt/wt), on the rheological, mechanical and sorption properties of urethane dimethylacrylate (UDMA)/triethylenglycol dimethacrylate (TEGDMA) (80/20wt/wt) resins blend. Silica nanoparticles (SiNP) were silanized with MPS or MPS+OTMS (50/50wt/wt) and incorporated in an UDMA-isopropanol mix to produce functionalized silica nanoclusters after evaporating the isopropanol. The effect of functionalized SiNC on resins rheological properties was determined by large and small deformation tests. Mechanical, thermal, sorption and solubility properties were evaluated for composite materials. The UDMA/TEGDMA (80/20wt/wt) resins blend with added SiNC (ca. 350nm) and functionalized with MPS showed a Newtonian flow behavior associated to their spheroidal shape, whereas the resins blend with nanoclusters silanized with MPS+OTMS (50/50wt/wt) (ca. 400nm) showed a shear-thinning behavior due to nanoclusters irregular shape. Composite materials prepared with bare silica nanoclusters showed lower compressive strength than functionalized silica nanoclusters. MPS functionalized nanoclusters showed better mechanical properties but higher water sorption than functionalized nanoclusters with both silane coupling agents, MPS and OTMS. The solvent evaporation method applied to functionalized nanoparticles showed to be an alternative way to the sinterization method for producing nanoclusters, which improved some dental composite mechanical properties and reduced water sorption. The shape of functionalized silica nanoclusters showed to have influence on the rheological properties of SiNC resin suspensions and the mechanical and sorption properties of light cured composites.


Specific antagonism of excitant amino acids in the isolated spinal cord of the neonatal rat.
The specificity of the neurodepressant actions of D-alpha-aminoadipate, alpha,epsilon-diaminopimelic acid, HA-966 (HAP) and Mg2+ has been investigated. On the isolated spinal cord of the neonatal rat, ventral root depolarizations produced by kainate, substance P, carbachol and noradrenaline were relatively unaffected by the same concentrations (0.25--1 mM) of the agents as those which reduced synaptic activity and ventral root depolarizations produced by N-methyl-D-aspartate (especially), L-aspartate and L-glutamate. The same or higher concentrations of the agents did not affect excitatory transmission in the isolated rat superior cervical ganglion. It is proposed that the agents specifically block synaptic transmission mediated by an excitatory amino acid.


Preliminary Phytochemical Studies.
Plants are the natural producers of medicinal agents like alkaloids, flavonoids, tannins, and phenolics. These phytocompounds alone or in combination act as a therapeutic agent in various disease complications. Various chemical reagents are used to determine the major phytochemicals present in plant parts. Protocols involved in screening of alkaloids, carbohydrates, glycosides, saponins, phytosterols, fixed oils, and fats are shown in this chapter.


Obstetric complications in carriers of the m.3243A>G mutation, a retrospective cohort study on maternal and fetal outcome.
The mitochondrial DNA m.3243A>G mutation is the most prevalent mutation causing mitochondrial disease in adult patients. Aside from some case reports, there are no studies on obstetric complications in a cohort of m.3243A>G carriers. We aimed to identify the prevalence of obstetric complications in a cohort of women carrying the m.3243A>G mutation. All female carriers of the m.3243A>G mutation known from our previous national inventory were sent a questionnaire regarding their obstetric history. Data were compared to national references. Data from the national inventory, including NMDAS (disease severity) scores and heteroplasmy levels in urinary epithelial cells (UEC) were used to stratify women. Sixty women participated, the mean age was 47 years (range 20-70), mean NMDAS was 14.6 (range 0-46), and mean heteroplasmy percentage in UEC was 19.9% (range 5-85%). Ninety-eight pregnancies in 46 women were reported. Twenty-three (25.3%) had a premature delivery and five of them (5.5%) had a gestation of ≤ 32 weeks and eleven of the women (12%) suffered from preeclampsia. No different heteroplasmy level was found in the women with preeclampsia. Nine pregnancies (11%) were complicated by gestational diabetes. Obstetric complications occur frequently in carriers of the m.3243A>G mutation. Proper guidance during pregnancies and early detection of possible obstetric complications are needed. As techniques to prevent transmission of mitochondrial mutations are studied it is important to know the possible complications patients may experience from the ensuing pregnancy.


Preparation and rearrangement of N-vinyl nitrones: synthesis of spiroisoxazolines and fluorene-tethered isoxazoles.
N-Vinyl nitrones derived from fluorenone have been prepared via a copper-mediated coupling between fluorenone oxime and vinyl boronic acids. These compounds undergo subsequent rearrangement and addition reactions that are distinct from the traditional [3 + 2] cycloaddition reactivity of nitrones. Thermal rearrangements of fluorenone N-vinyl nitrones give spiroisoxazolines, while treatment with alkynes provides fluorene-tethered isoxazoles. The scope and limitations of the preparation of fluorenone N-vinyl nitrones and their subsequent rearrangement and addition reactions are discussed.


Comparison of MALDI-TOF mass spectrometry and rpoB gene sequencing for the identification of clinical isolates of Aeromonas spp.
Aeromonas spp., widely present in rivers and soil, cause mild gastroenteritis, severe septicemia, and soft tissue infections in humans. Treatment of these infections require accurate identification of pathogenic Aeromonas spp. However, identification at the species level using conventional methods is highly challenging. In this study, we aimed to compare the accuracy of two different approaches developed for bacterial identification: (i) housekeeping gene sequencing (rpoB) in conjunction with phylogenetic analysis and (ii) matrix-assisted laser desorption ionization mass spectrometry-time of flight (MALDI-TOF MS) (MALDI Biotyper and VITEK MS), for differentiating Aeromonas spp. We analyzed 58 Aeromonas isolates recovered from patients at different medical institutions in Japan using both identification methods. The rpoB sequencing method was the most accurate, identifying all Aeromonas isolates at the species level. Meanwhile, the MALDI Biotyper system correctly identified 53 (91.4%) isolates at the genus level and an additional 30 (51.7%) at the species level. The VITEK MS system correctly identified 58 (100%) isolates at the genus level and an additional 34 (58.6%) at the species level. Thus, MALDI Biotyper and VITEK MS accurately identified isolates at the genus level, but differences were found in the accuracy of identification of species. However, the low cost and ease of analysis make MALDI-TOF MS-based methods strong candidates for use in clinical laboratories that require easy-to-use identification methods.


Y chromosome mosaicism in the gonads, but not in the blood, of a girl with the Turner phenotype and virilized external genitalia.
We describe a girl with virilized external genitalia and phenotypic features of Turner syndrome whose blood karyotype is 45,X. The presence of dysgenetic testicular tissue was confirmed by pathology. Using PCR and primers for the distal long arm, centromere and short arm of the Y chromosome, Y chromosome material was detected in her gonads but not in blood.


[Changes in the content of lipids and biologically active substances under the influence of the ultraviolet irradiation of the blood in experimental peritonitis].
In dogs with experimental peritonitis, accumulation of toxic metabolic products caused impairment in lipid metabolism. With reduction in pronouncement of endotoxemia under the influence of ultra-violet irradiation, the blood level of biologically active substances decreased, the process of lipolysis was suppressed.


Oral-bacterial-induced arterial and venous thrombus in rats: Pathological and immunological studies.
Our study investigated the pathological outcome of experimental thrombi that incorporate oral bacteria. A small artery and vein in the rats' groins were injected with a solution containing periodontal bacteria Porphyromonas gingivalis and followed up for 28 days. In all, 18 limbs of nine male rats (500-650 g) were used for the arterial study, and eight limbs of four rats were used for the veins. Two densities of the bacterial solution and two arterial thicknesses sizes were used in the arterial study. Both proximal and distal arteries and veins were ligated loosely using a monofilament nylon suture before bacterial suspensions or control solutions were injected into the ligated vessels. After 7, 14-18, and 28 days, the rats were sacrificed. Pathology and immunohistochemistry were performed. All specimens exhibited thrombus formation and an acute inflammation reaction with granulocytes at 7 days and then settled down to chronic fibrous change with plasma cells or macrophages at 28 days in the arterial thrombus. CD3 (Pan T-cells), CD79a (Pan B cells in the rats), and IgG were observed in the process of the healing of the arterial thrombus. Venous changes showed relatively clear recanalization that appeared at 7 days, which is slightly different from the artery. Granulocytes were present from 7 to 28 days. Periodontal bacteria act as an inflammatory core in the vessels, but not as an infectious agent, in our experiments, because of their low ability to invade tissues.


Tumor-associated CD163+ macrophage as a predictor of tumor spread through air spaces and with CD25+ lymphocyte as a prognostic factor in resected stage I lung adenocarcinoma.
Lung cancer can spread in numerous ways, including spread through air spaces (STAS). A high number of CD68+ tumor-associated macrophages (TAMs), which creates a favorable microenvironment for tumor progression, is an independent predictor of increased STAS rate and is used as a pan-macrophage marker, whereas CD163 is used as an M2 macrophage marker. A high number of CD25+ tumor-infiltrating lymphocytes (TILs) is associated with the frequency of STAS. This study investigated the influence of M2 macrophages and CD25+ TILs on STAS and postoperative recurrence in patients with stage I lung adenocarcinoma who underwent curative resection. We analyzed data from 485 patients with stage 0-I lung adenocarcinoma who underwent resection between 1999 and 2016. Tissue microarrays were constructed, and immunohistochemical analysis was performed for CD3, CD4, CD8, CD45RO, CD25, CD20, CD68, and CD163. Three tumor areas with the highest density of immune cells were photographed, and the immune cells were quantified. Associations between variables were analyzed using chi-square tests and Mann-Whitney U tests. Recurrence-free probability (RFP) was analyzed using log-rank tests and Cox proportional hazards models. CD163+ TAMs were identified as an independent predictor of a higher rate of STAS (P < 0.001). Analysis of biologically relevant immune cell combinations revealed that patients with high CD25+ TILs and high CD163+ TAMs had a significantly lower RFP (5-year RFP, 74%) than those with other combinations of CD25 and CD163 (5-year RFP, 90%; P < 0.001). Multivariate analysis showed that high CD25+/high CD163+ immune cell infiltration was an independent predictor of RFP. We demonstrated that a higher density of M2 macrophages is an independent predictor of a higher STAS incidence. A high CD25+/high CD163+ immune cell infiltration ratio is a significant prognostic factor for stage 0-I lung adenocarcinoma.


Graft side-mismatching for single-lung transplantation does not affect outcomes: role of the pre-operative quantitative lung perfusion scan.
There are concerns about which lung to explant during single-lung transplantation (SLT). Traditionally, a quantitative lung perfusion scan (QLPS) is performed, and the better-perfused lung is retained. Occasionally, there is transplantation with graft "side-mismatching," where the less-well-perfused lung is retained. We performed a retrospective study of patients undergoing SLT at our institution to evaluate the effects of side-mismatching (according to the QLPS) on graft performance and outcome. We defined graft side-mismatching with a prospectively designed formula using baseline QLPS, and defined patients as either side-matched or side-mismatched. Data on mortality, requirement for cardiopulmonary bypass, relative graft perfusion, lung function and exercise capacity were obtained from institutional databases and patients' files. In a cohort of 114 patients, we defined 97 as having received a side-matched SLT and 17 as having received a side-mismatched graft. After lung transplantation, forced expiratory volume in 1 second (FEV(1)) and exercise capacity improved in both groups (p < 0.001). Patients with mismatched lungs had significantly higher relative graft perfusion post-operatively (p = 0.0012). There was no significant difference between the two groups (matched vs mismatched) in mortality, physiologic parameters and need for cardiopulmonary bypass. There is no apparent risk to the patient when a side-mismatched lung graft is transplanted. We conclude that side-mismatched lung transplantation appears to be feasible when required.


Voltammetric carbon paste electrodes monitor uric acid and not 5-HIAA at the 5-hydroxyindole potential in the rat brain.
Changes in the height of peak 2 obtained using linear sweep voltammetry and carbon paste electrodes chronically implanted in discrete brain regions of the unrestrained rat were measured under a variety of conditions; in the past this peak has been attributed to the oxidation of 5-hydroxyindoleacetic acid (5-HIAA). Unilateral 5,7-dihydroxytryptamine (5,7-DHT) lesions of the medial forebrain bundle reduced the 5-HIAA content of the striatum and hippocampus to 10% of the unlesioned side, but did not alter the height of peak 2 recorded in these regions. In contrast, microinfusion of uricase beside striatial electrodes reduced the height of peak 2 by 96%; systemic amphetamine-induced increases in the height of the peak were also prevented by this enzyme. These results indicate that uric acid, and not 5-HIAA, is mainly responsible for peak 2, and that changes in the height of this peak reflect changes in the extracellular concentration of uric acid.


Influence of the drug deformation behaviour on the predictability of compressibility and compactibility of binary mixtures.
Various studies investigate the predictability of the compressibility and compactibility of tablet formulations based on the behaviour of the pure materials. However, these studies are limited to a few materials so far probably because of the complexity of the powder compaction process. One approach preventing the excessive increase in complexity is the extension of the investigations from pure materials to binary powder mixtures. The focus of this study is on the predictability of the compressibility and compactibility of binary mixtures consisting of an active pharmaceutical ingredient (API) and the excipient microcrystalline cellulose. Three APIs with markedly different deformation behaviour were used. The API concentration and type are systematically varied. For all three material combinations it is found that the in-die compressibility of the binary mixtures can be precisely predicted based on the characteristic compression parameters of the raw materials using the extended in-die compression function in combination with a volume-based linear mixing rule. Since the tablet porosity (out-of-die) also follows a linear mixing rule, the predictability can be further extended using the method of Katz et al. In contrast, the influence of the API concentration on compactibility or rather on tablet tensile strength is non-linear and strongly dependent on the deformation behaviour of the API, making the predictability more difficult. Neither the approach of Reynolds et al. nor this of Kuentz and Leuenberger are able to predict the compactibility when clear deviations from a linear mixing rule appear.


Understanding Users in the 'Field' of Medications.
The numbers of medicinal drugs available for human consumption have increased rapidly in the past several decades, and physician prescribing practices reflect the growing reliance on medicines in health care. However, the nature of medicines-as-technology makes problematic taken-for-granted relationships among actors involved in the delivery, or who are the recipients of medicines-reliant health care. In this article, I situate the medicine user in the 'field' of medications-where interests, actions and outcomes are continually negotiated among and between the various players-physicians, pharmacists, government regulatory bodies, the pharmaceutical industry and users of medicines. The objective of the paper is to illuminate the complex context in which the medicine-user-the target of the pharmacy profession's service to the public-accesses and uses medicines.


First-line axitinib therapy is less effective in metastatic renal cell carcinoma with spindle histology.
Axitinib, a vascular endothelial growth factor receptor-tyrosine kinase inhibitor, will be used in combination first-line therapies against metastatic renal cell carcinoma (mRCC), but its effects as a first-line monotherapy are unclear. Thus, we aimed to elucidate pretreatment clinical factors that predict the prognosis of patients with mRCC receiving first-line axitinib therapy. We enrolled 63 patients with mRCC treated with axitinib as first-line therapy between Nov. 2003 and Jul. 2018. Progression-free survival (PFS) and overall survival (OS) were assessed using the Wald χ2 statistic in Cox proportional hazards regression. Median patient age was 67 (range: 25-85) years. Seven (11.1%) patients were classified as being at favorable risk, 33 (52.4%) at intermediate risk, and 23 (36.5%) at poor risk according to the International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk classification system. Median follow-up duration after axitinib initiation was 14 (range: 1-72) months. Median PFS and OS were 18 months and 65 months, respectively. Cox regression analyses of clinical predictors revealed that high C-reactive protein (CRP) levels were significantly correlated with shorter PFS [hazard ratio (HR), 1.63; 95% confidence interval (CI) 1.7-4.0)], whereas spindle cells and poor IMDC risk scores were related to worse OS (HR, 2.87 and 2.88, respectively; 95% CI 1.4-11.0 and 1.1-8.5, respectively). Thus, patients with mRCC and spindle histology or poor IMDC risk scores had worse OS, and those with high CRP levels had shorter PFS in first-line axitinib treatment. Other therapies might be more suitable for initial management of such patients.


Monitoring membrane viscosity in differentiating stem cells using BODIPY-based molecular rotors and FLIM.
Membrane fluidity plays an important role in many cell functions such as cell adhesion, and migration. In stem cell lines membrane fluidity may play a role in differentiation. Here we report the use of viscosity-sensitive fluorophores based on a BODIPY core, termed "molecular rotors", in combination with Fluorescence Lifetime Imaging Microscopy, for monitoring of plasma membrane viscosity changes in mesenchymal stem cells (MSCs) during osteogenic and chondrogenic differentiation. In order to correlate the viscosity values with membrane lipid composition, the detailed analysis of the corresponding membrane lipid composition of differentiated cells was performed by time-of-flight secondary ion mass spectrometry. Our results directly demonstrate for the first time that differentiation of MSCs results in distinct membrane viscosities, that reflect the change in lipidome of the cells following differentiation.


DHA alleviates high glucose-induced mitochondrial dysfunction in Oreochromis niloticus by inhibiting DRP1-mediated mitochondrial fission.
Dynamin-related protein 1 (DRP1) is a key regulator in the maintenance of mammalian glucose homeostasis, but the relevant information remains poorly understood on aquatic animals. In the study, DRP1 is formally described for the first time in Oreochromis niloticus. DRP1 encodes a peptide of 673 amino acid residues that contained three conserved domains: a GTPase domain, a dynamin middle domain and a dynamin GTPase effector domain. DRP1 transcripts are widely distributed in all of the detected seven organs/tissues, and the highest mRNA levels in brain. High-carbohydrate (45 %) fed fish showed a significant upregulation of liver DRP1 expression than that of control (30 %) group. Glucose administration upregulated liver DRP1 expression, with peak values observed at 1 h; then its expression returned to the basal value at 12 h. In the in vitro study, DRP1 over-expression significantly decreased mitochondrial abundance in hepatocytes. DHA significantly increased mitochondrial abundance, transcriptions of mitochondrial transcription factor A (TFAM) and mitofusin 1 and 2 (MFN1 and MFN2) and complex II and III activities of high glucose-treated hepatocyte, whereas the opposite was true for DRP1, mitochondrial fission factor (MFF) and fission (FIS) expression. Together, these findings illustrated that O. niloticus DRP1 is highly conserved, and it participated in glucose control of fish. DHA could alleviate high glucose-induced mitochondrial dysfunction of fish by inhibiting DRP1-mediated mitochondrial fission.


Molecular characterization and clinical outcomes in EGFR-mutant de novo MET-overexpressed advanced non-small-cell lung cancer.
Approximately 2%-8% of non-small-cell lung cancer (NSCLC) harbors concurrent epidermal growth factor receptor (EGFR) sensitizing mutation and mesenchymal-epithelial transition factor (MET) amplification prior to EGFR-tyrosine kinase inhibitor (EGFR-TKI) therapy. This study aimed to investigate the optimal first-line therapeutic options for patients with concurrent EGFR-mutant, MET-overexpressed/amplified advanced NSCLC. A total of 104 treatment-naïve patients with EGFR-mutant de novo MET-overexpressed advanced NSCLC were identified using immunohistochemistry and stratified to four groups according to treatment regimen: EGFR-TKI monotherapy (n = 48), EGFR-TKI combined with either crizotinib (n = 9) or chemotherapy (n = 12), and chemotherapy (n = 35). A subpopulation of 28 patients was also tested with next-generation sequencing (NGS). Objective response rate (ORR) and progression-free survival (PFS) outcomes were analyzed according to treatment strategies and molecular features. All the patients (n = 104) achieved ORR of 36.5% and median PFS (mPFS) of 7.0 months. Baseline clinicopathologic characteristics were similar among the four treatment groups. Compared with chemotherapy, EGFR-TKI monotherapy or EGFR-TKI combination therapy achieved significantly higher ORR (P < 0.001) and longer mPFS (P = 0.003). No ORR or PFS difference was observed between EGFR-TKI monotherapy and combination therapy. In the NGS-identified population (n = 28), patients who received EGFR-TKI plus crizotinib (n = 9) achieved similar ORR (88.9% versus 57.9%, P = 0.195) and mPFS (9.0 versus 8.5 months, hazard ratio 1.10, 95% confidence interval 0.43-2.55, P = 0.45) than those who received EGFR-TKI monotherapy (n = 19), regardless of MET copy number status. Grade 3/4 rashes were significantly more among patients who received EGFR-TKI plus crizotinib (P = 0.026). Our findings provided clinical evidence that patients with concurrent EGFR sensitizing mutation and de novo MET amplification/overexpression could benefit from first-line EGFR-TKI monotherapy.


Comparison between fractionation and bioavailability of trace elements in rhizosphere and bulk soils.
Rhizosphere is a microbiosphere and has quite different chemical, physical and biological properties from bulk soils. A greenhouse experiment was performed to compare the difference of fractionation and bioavailability of trace elements Cr, Ni, Zn, Cu, Pb and Cd between rhizosphere soil and bulk soil. In the meantime, the influence of air-drying on the fractionation and bioavailability was also investigated by using wet soil sample as a control. Soils in a homemade rhizobox were divided into four zones: rhizosphere, near rhizosphere, near bulk soil and bulk soil zones, which was designated as S1, S2, S3 and S4. Elemental speciations were fractionated to water soluble, exchangeable and carbonate bound (B1), Fe-Mn oxide bound (B2), and organic and sulfide bound (B3) by a sequential extraction procedure. Speciation differences were observed for elements Cr, Ni, Zn, Cu, Pb and Cd between the rhizosphere and bulk soils, and between the air-dried and wet soils as well. The concentrations of all six heavy metals in fraction B1 followed the order of S2 > S3 > S1 > S4 and for B2, the order was S2 > S3 S4 > S1. For B3, the order was S1 > S3 S4 > S2, while for Cd the order was S2 > S3 approximately/= S4 > S1. The air-drying increased elemental concentration in fractions B1 and B2 by 20-50% and decreased in fraction B3 by about 20-100%. Correlation analysis also indicated that the bioavailability correlation coefficient of fraction B1 in rhizosphere wet soil to plants was better than that between either air-dried or nonrhizosphere soils. Therefore, application of rhizosphere wet soils should be recommended in the future study on the speciation analysis of trace elements in soils and bioavailability.


A Method to Identify Potential Prognostic Markers Across Distinct Tumor Types.
The identification of novel biomarkers in cancer patients often requires both survival and gene expression analyses. The Kaplan-Meier survival analysis is one of the most common methods to assess the fraction of subjects living for a certain amount of time.Here, we describe a method for researchers to identify potential prognostic markers across distinct tumor types. We utilize The Cancer Genome Atlas (TCGA) as this is one of the most extensive and successful cancer genomics programs to date that includes expression data and clinical follow-up information for up to 33 distinct tumor types. Nevertheless, the method described here can also be applied to any open-source dataset where the RNA expression and clinical outcome are provided.We provide detailed practical instructions and advices for investigators to be able to successfully identify prognostic markers in cancer patients.