Some correlations between light transmission changes and some commonly used in vitro assays for the assessment of platelet concentrates.
This study aims to relate the duration of light transmission changes (LTCs) through platelet concentrates (PCs) to the results of a panel analytical procedures, many of which have correlated with platelet in vivo viability. LTCs of 16 PCs stored in P1-732 containers were studied over 10 d. On d 1, platelet counts and total PC ATP content were determined. On d 7, determination of the following parameters was carried out: pH, bicarbonate, the rate of oxygen consumption, the dispersion of the size distribution, the total platelet ATP content, the extent of shape change (ESC), the osmotic reversal reaction and extracellular lactate dehydrogenase (LDH) level. In addition, the rate of lactate production was determined over storage. A second series consisting of 18 PCs stored in P1-146 containers was monitored over 7 d. Mean platelet volume (MPV) was determined on d 3 and 7 and ESC was analysed on d 3. LTCs correlated significantly with pH, bicarbonate, the rate of oxygen consumption, increased platelet size distribution and decreased ESC. In addition, significant correlations were found when comparing LTCs with MPV(s). This study demonstrates that LTCs are closely related to parameters reflecting platelet metabolism.


Effects of cholesterol feeding on synthesis and metabolism of cholesterol and bile acids in germfree rats.
The aim of the present investigation was to study the influence of cholesterol feeding on cholesterol synthesis and fecal excretion of bile acids in germfree rats. Four germfree rats were fed a basal diet containing 0.004% cholesterol and four germfree rats received the same diet supplemented with 0.4% cholesterol for 2 weeks. Cholesterol synthesis was studied by assaying the HMG CoA reductase activity in the liver microsomal fraction. Cholesterol feeding decreased the HMG CoA reductase activity from 28.5 +/- 6.6 (mean +/- SEM) to 9.1 +/- 0.7 pmol/mg protein per min. In another experiment four germfree rats received the basal diet and four germfree rats the cholesterol-enriched diet. After 6 weeks feces were collected in two 4-day pools for analysis of bile acids. The main fecal bile acids were cholic acid and beta-muricholic acid (a metabolite of chenodeoxycholic acid), comprising more than 95% of total bile acids. Cholic acid was increased from 3.9 +/- 0.2 to 9.9 +/- 1.2 mg/kg body weight per day and beta-muricholic acid from 6.6 +/- 0.5 to 21.8 +/- 3.1 mg/kg body weight per day. The percentage of cholic acid decreased from 37.1 +/- 1.1 to 31.2 +/- 1.0%. In conclusion, germfree rats like conventional rats have the ability to compensate for an increased input of dietary cholesterol by inhibition of cholesterol synthesis and stimulation of bile acid synthesis. The synthesis of chenodeoxycholic acid (implied from the fecal excretion of beta-muricholic acid) is stimulated to a greater extent than that of cholic acid.


Organocatalytic asymmetric synthesis of β,β-diaryl ketones via one-pot tandem dehydration/1,6-addition/decarboxylation transformation of β-keto acids and 4-hydroxybenzyl alcohols.
Herein we describe an organocatalytic protocol for the asymmetric synthesis of β,β-diaryl ketones. Under the catalysis of a chiral phosphoric acid, 4-hydroxybenzyl alcohols underwent dehydration to form para-quinone methides, which reacted with β-keto acids in 1,6-addition reactions. Upon treatment with Et3N in one-pot, decarboxylation proceeded to provide the desired chiral ketones in nearly quantitative yields with high enantioselectivities.


Interleukin-6 (IL-6) is an intermediate in IL-1-induced proliferation of leukemic human megakaryoblasts.
We have examined the in vitro effects of recombinant human (rh) interleukin-1 (IL-1) on the growth of purified megakaryoblasts obtained from patients with acute megakaryoblastic leukemia. We demonstrate that both IL-1 alpha and IL-1 beta treatment of these cells led to stimulation of DNA synthesis (as shown by increase of 3H-thymidine incorporation up to 35-fold) and also resulted in colony formation of leukemic megakaryoblasts. However, the stimulatory effect of IL-1 was dependent on endogenous production of IL-6, because addition of neutralizing monoclonal antibody (MoAb) to IL-6 abrogated the stimulatory activity of IL-1. In contrast, neutralizing MoAbs to granulocyte (G)-colony stimulating factor (CSF), granulocyte-macrophage (GM)-CSF, and macrophage (M)-CSF failed to counteract the growth-enhancing effects of IL-1. Leukemic megakaryoblasts accumulated IL-6 mRNA and released IL-6 protein into their culture supernatant when exposed to rh IL-1 but failed to disclose transcripts for G-, GM-, and M-CSF under these conditions. Analysis of IL-6 receptor (IL-6R) transcript levels demonstrated that megakaryoblasts constitutively expressed IL-6R mRNA and that these transcripts are down-regulated to undetectable levels upon exposure to IL-1 and IL-6. Increase of 3H-thymidine incorporation by megakaryoblasts could be duplicated by exogenous IL-6 that could be blocked by neutralizing MoAb to IL-6. In conclusion, our results suggest that leukemic megakaryoblasts could produce and secrete IL-6, and express IL-6R, and that the growth-enhancing effect of IL-1 on these cells is indirect, via production of IL-6 by leukemic cells.


Spatio-temporal evolution of human neural activity during visually cued hand movements.
Making hand movements in response to visual cues is common in daily life. It has been well known that this process activates multiple areas in the brain, but how these neural activations progress across space and time remains largely unknown. Taking advantage of intracranial electroencephalographic (iEEG) recordings using depth and subdural electrodes from 36 human subjects using the same task, we applied single-trial and cross-trial analyses to high-frequency iEEG activity. The results show that the neural activation was widely distributed across the human brain both within and on the surface of the brain, and focused specifically on certain areas in the parietal, frontal, and occipital lobes, where parietal lobes present significant left lateralization on the activation. We also demonstrate temporal differences across these brain regions. Finally, we evaluated the degree to which the timing of activity within these regions was related to sensory or motor function. The findings of this study promote the understanding of task-related neural processing of the human brain, and may provide important insights for translational applications.


Exploring students' experience of training for counselling skills and the impact on practice.
The aim of this study was to explore participants' experience of an accredited counselling skills course and their perception of its impact on practice. A qualitative design was used utilizing a mixed-method approach. Data were collected using semistructured interviews and the repertory-grid technique. Data were analysed using thematic content analysis (interviews) and the repertory grids were analysed qualitatively using a four-stage procedure. A convenience sample of six healthcare professionals undertaking a counselling skills course completed both interviews. The biggest impact of the course was in the area of self-awareness and skill development. The development of attentive listening, both in terms of objective and subjective findings, was evident. Students perceived that personal qualities, such as warmth and caring, were essential. All participants identified that the group were supportive and that this enhanced learning. The students changed their practice by improving their skills, increasing their awareness and by developing deeper therapeutic relationships with patients and their carers. This research adds to the body of knowledge in palliative care. It identifies key elements in the learning of counselling skills and recognizes the impact of these skills on improving the care of palliative care patients and families.


D-CHART: A Novel Method of Measuring Metamorphopsia in Epiretinal Membrane and Macular Hole.
Metamorphopsia is common in macular disease. Current techniques for measuring metamorphopsia require good vision or costly equipment. The authors report a method that uses printed cards. The cards have a grid of squares arranged in a ring around fixation. There are four rings, at different distances from fixation, divided into eight sectors. The separation of the grid elements ranges from 0.4° to 1.8°. Subjects indicate in which sector lines of squares are distorted. The sum of the maximum separation perceived as distorted in each sector gives the total metamorphopsia score. Thirty-three eyes with epiretinal membrane and 29 eyes with macular hole were tested. Twenty-four eyes were tested again after surgery. In 18 subjects, the preoperative test was performed twice to assess repeatability. The median preoperative total metamorphopsia score was 10.2 for macular hole and 5.2 for epiretinal membrane. After surgery, the median total metamorphopsia score was 0.5 for macular hole and 0.45 for epiretinal membrane. Test-retest results showed good correlation. Improvement in metamorphopsia did not correlate with change in visual acuity. Measurement of metamorphopsia may be useful in the management of macular hole and epiretinal membrane. D-charts are a simple and inexpensive method of quantifying metamorphopsia that can be used in a clinical setting.


The functional domains of bacteriophage t4 terminase.
The packaging of double-stranded genomic DNA into some viral and all bacteriophage capsids is driven by powerful molecular motors. In bacteriophage T4, the motor consists of the portal protein assembly composed of twelve copies of gene product 20 (gp20, 61 kDa) and an oligomeric terminase complex composed of gp16 (18 kDa) and gp17 (70 kDa). The packaging motor drives the 171-kbp T4 DNA into the capsid utilizing the free energy of ATP hydrolysis. Evidence suggests that gp17 is the key component of the motor; it exhibits ATPase, nuclease, and in vitro DNA-packaging activities. The N- and C-terminal halves of gp17 were expressed and purified to homogeneity and found to have ATPase and nuclease activities, respectively. The N-terminal domain exhibited 2-3-fold higher Kcat values for gp16-stimulated ATPase than the full-length gp17. Neither of the domains, individually or together, exhibited in vitro DNA-packaging activity, suggesting that communication between the domains is essential for DNA packaging. The domains, in particular the C-terminal domain or a mixture of both the N- and C-terminal domains, inhibited in vitro DNA packaging that is catalyzed by full-length gp17. In conjunction with genetic evidence, these data suggest that the domains compete with the full-length gp17 for binding sites on the portal protein. A model for the assembly of the T4 DNA-packaging machine is presented.


Epidermal growth factor induces tyrosine phosphorylation, membrane insertion, and activation of transient receptor potential channel 4.
Various members of the canonical family of transient receptor potential channels (TRPCs) exhibit increased cation influx following receptor stimulation or Ca(2+) store depletion. Tyrosine phosphorylation of TRP family members also results in increased channel activity; however, the link between the two events is unclear. We report that two tyrosine residues in the C terminus of human TRPC4 (hTRPC4), Tyr-959 and Tyr-972, are phosphorylated following epidermal growth factor (EGF) receptor stimulation of COS-7 cells. This phosphorylation was mediated by Src family tyrosine kinases (STKs), with Fyn appearing to be the dominant kinase. In addition, EGF receptor stimulation induced the exocytotic insertion of hTRPC4 into the plasma membrane dependent on the activity of STKs and was accompanied by a phosphorylation-dependent increase in the association of hTRPC4 with Na(+)/H(+) exchanger regulatory factor. Furthermore, this translocation and association was defective upon mutation of Tyr-959 and Tyr-972 to phenylalanine. Significantly, inhibition of STKs was concomitant with a reduction in Ca(2+) influx in both native COS-7 cells and hTRPC4-expressing HEK293 cells, with cells expressing the Y959F/Y972F mutant exhibiting a reduced EGF response. These findings represent the first demonstration of a mechanism for phosphorylation to modulate TRPC channel function.


Enhanced Neurobehavioral Outcomes of Action Observation Prosthesis Training.
Background Previous studies have demonstrated improved neurobehavioral outcomes when prosthesis users learn task-specific behaviors by imitating movements of prosthesis users (matched limb) compared with intact limbs (mismatched limb). Objective This study is the first to use a unique combination of neurophysiological and task performance methods to investigate prosthetic device training strategies from a cognitive motor control perspective. Intact nonamputated prosthesis users (NAPUs) donned specially adapted prosthetic devices to simulate the wrist and forearm movement that persons with transradial limb loss experience. The hypothesis is that NAPUs trained with matched limb imitation would show greater engagement of parietofrontal regions and reduced movement variability compared with their counterparts trained with a mismatched limb. Methods Training elapsed over 3 days comprised alternating periods of video demonstration observation followed by action imitation. At the beginning and end of the training protocol, participants performed a cued movement paradigm while electroencephalography and electrogoniometry data were collected to track changes in cortical activity and movement variability, respectively. Results Matched limb participants showed greater engagement of motor-related areas while mismatched limb participants showed greater engagement of the parietooccipital system. Matched limb participants also showed lower movement variability. Conclusions These results indicate that the type of limb imitated influences neural and behavioral strategies for novel prosthetic device usage. This finding is important, as customary prosthetic rehabilitation with intact therapists involves mismatched limb imitation that may exacerbate challenges in adapting to new motor patterns demanded by prosthesis use.


Kläui ligand thin films for rapid plutonium analysis by alpha spectrometry.
As part of a nuclear forensics capability, rapid and effective methods to analyze for plutonium and other actinide metals are needed. A key requirement of these methods is that they afford a high chemical yield while still providing isotopic information necessary for forensic evaluation. Toward this objective, a new method for binding plutonium for analysis by alpha spectrometry has been developed. Thin films of Kläui-type tripodal oxygen donor ligands were prepared by spin-casting solutions onto glass substrates. Three different ligands were evaluated for plutonium binding, and the best results were obtained using the ethyl-substituted complex Na[Cp*Co(P(O)(OEt)2)3], which bound 80-88% of the dissolved Pu under equilibrium conditions. The thin films are simple and inexpensive to prepare and exhibit excellent alpha spectral resolution, having line widths of ~33 keV. The method has been successfully applied to analyze for plutonium in both an archived nuclear debris sample and a certified environmental soil sample. The results obtained from the soil analysis are in good agreement with the certified values, demonstrating the effectiveness of the method for rapid plutonium analysis.


Quality of life and disease-specific functioning of patients with anorectal malformations or Hirschsprung's disease: a review.
The objective of this review was to examine disease-specific functioning (ie, faecal functioning and disease-related psychosocial problems) and quality of life (QoL)in patients with anorectal malformations (ARMs) or Hirschsprung's disease across different developmental stages. A search on PubMed yielded 22 eligible studies which were analysed. Patients with ARM or Hirschsprung's disease report slightly more QoL problems than comparison groups. As compared with adolescents, children reported better QoL but worse disease-specific functioning. Relationships between disease-specific functioning and QoL remain unclear. Therefore, to disentangle the complex relationship between the faecal functioning of these patients and their QoL through childhood, adolescence and adulthood, longitudinal studies should examine disease-specific functioning with validated QoL questionnaires that include age-specific versions.


Emotion regulation strategies moderate the impact of negative affect induction on alcohol craving in college drinkers: an experimental paradigm.
Objectives: Observational studies suggest emotion regulation (ER) as a potential treatment target for problematic college drinking. The primary aim of this laboratory study was to determine whether trait ER strategies would moderate the impact of negative affect induction on alcohol craving in college drinkers. Methods: Participants were randomly assigned to a neutral (n = 74) or a negative affect induction (n = 76) and reported their craving after the affect inductions. Results: Greater use of drinking to cope and less use of cognitive reappraisal predicted greater alcohol craving after the negative affect induction, but not after the neutral condition. In contrast, emotion suppression did not predict alcohol craving in either condition. Conclusion: Our results highlight the role of ER tendencies-particularly the benefits of cognitive reappraisal-on alcohol craving when experiencing emotional distress. Therefore, ER strategies may be an important target for college drinkers to prevent and reduce problematic drinking.


Total synthesis of the L-hexoses.
Enantiomerically pure polyhydroxylated natural products are synthesized by using a reiterative two-carbon extension cycle consisting of four steps. The generality and efficiency of this methodology are demonstrated in the total synthesis of all eight L-hexoses.


Radial profile measurements of plasma pressure-like fluctuations with the heavy ion beam diagnostic on the tokamak ISTTOK.
The Heavy Ion Beam Diagnostic (HIBD) on the tokamak ISTTOK (Instituto Superior Técnico TOKamak) has been modified, in terms of signal conditioning, to measure the local fluctuations of the neσ1,2(Te) product (plasma density times the effective ionization cross-section) along the tokamak minor diameter, in 12 sample volumes in the range of -0.7a < r < 0.7a, with a maximum delay time of 1 μs. The corresponding signals show high correlation with the magnetic Mirnov coils in the characteristic MHD frequency range of ISTTOK plasmas and enable the identification of tearing modes. This paper describes the HIBD signal conditioning system and presents a preliminary analysis of the radial profile measurements of local neσ1,2(Te) fluctuations.


Outcome determinants for transformed indolent lymphomas treated with or without autologous stem-cell transplantation.
Transformation of indolent lymphomas (IL) to an aggressive histology (TIL) often results in a rapid clinical course, treatment refractoriness and shortened survival. Although rituximab-containing regimens (R-chemo) have become standard of care in CD20-positive TIL, the role of autologous stem-cell transplantation (ASCT) is still debated. The purpose of this study was to determine whether the outcome of TIL patients improved if they, at transformation, also received ASCT. Furthermore, we investigated the outcome of cases with histologically low- and high-grade components diagnosed either simultaneously or after a period of overt indolent disease. We also analyzed, whether prior rituximab treatment during the indolent course of the disease affected outcome after transformation. Eighty-five patients (≤68 years) with histologically confirmed TIL were included. Five-year overall (OS) and progression-free survival (PFS) were calculated. Selected parameters were tested in a multivariate analysis. All analyses were conducted on three cohorts: (i) whole cohort (all TIL), (ii) patients with co-existing evidence of both indolent and aggressive histology at diagnosis (Composite/discordant TIL) and (iii) patients transformed after prolonged prior indolent disease (sequential TIL). Fifty-four patients (64%) received ASCT consolidation and 31 (36%) did not. Within the 'all TIL' cohort, the 5-year OS and PFS for R-chemo + ASCT versus R-chemo alone, were 67% versus 48% (P = 0.11) and 60% versus 30% (P = 0.02), respectively. Furthermore, in 'Composite/discordant TIL' R-chemo + ASCT showed no impact on OS (76% versus 67%; P = 0.66) or PFS (71% versus 62%; P = 0.54). Conversely, R-chemo + ASCT improved the outcome of 'sequential TIL' (OS 62% versus 36%; P = 0.07; PFS 53% versus 6%; P = 0.002), regardless of prior rituximab therapy. The beneficial effect of ASCT was significantly higher in patients who had not received rituximab at IL stage. ASCT improved the outcome in sequential, but not composite/discordant TIL. The beneficial impact of ASCT was greater in patients, who were rituximab-naïve at transformation.


Retinal artery occlusion in a patient with factor V Leiden and prothrombin G20210A mutations.
Retinal artery occlusion is rare in young adults, and may be associated with hereditary thrombophilia. We present a 19-year-old male who was evaluated for central retinal artery occlusion and found to be homozygous for the factor V Leiden mutation and heterozygous for the prothrombin G20210A mutation. Anterior chamber paracenthesis resulted in dramatic improvement, but recurring loss of vision necessitated repeated paracenthesis and the addition of aspirin to standard anticoagulation treatment. The literature concerning hereditary thrombophilia and retinal artery occlusion is reviewed, and the synergistic effect of multiple risk factors is emphasized. Screening for hereditary thrombophilia should be considered for young people presenting with unexplained retinal artery occlusion.


[Inguinal hernia in the elderly. Indications, techniques, results].
Inguinal hernia in the elderly is a quite common problem in general surgery. Surgical treatment must be aware of the cardiovascular and respiratory pathology these patients are often affected by and, on the other side, the goal of surgery is a quick functional recovery after the operation using the "tension free" technique and, whenever possible, local anesthesia. In this paper personal experience on inguinal hernioplasty in elderly patients (>65 years) is presented; the aim is a better definition of the indications to surgery and anesthesia and verify the results of surgical treatment. From January 1999 to December 2002 our group operated 317 patients with inguinal hernia; 123 patients were over 65 (119 male, 4 female, age from 65 to 101 years, mean 74); in this group 111 Lichtenstein and 12 Trabucco HPL were performed. There were 89 primitive hernias, 17 recurrences and 17 emergency operation. Local anesthesia was the method of choice except for obese patients, obstruction or strangulation, or patient's preference. In giant hernia, or when a large sac was found, the Wantz technique was performed (13 patients); a suction drainage was used if a large space in the inguinoscrotal area remained after hernia reduction (9 patients). Local anesthesia was used in 87 patients, spinal in 31 and 5 in general; there were no postoperative deaths. Local complications were: 4 inguinal hematomas, 2 hematomas and 2 transient edemas of the scrotum, 1 wound infection, 1 sieroma of the wound and 1 scrotal sieroma (9%); all patients recovered after few days of medical therapy. No cases of ischemic orchitis occurred. Inguinal herniorraphy can be safely performed, in elderly patients; according to personal experience local anesthesia proved to be a safe technique, also in patients with cardiocirculatory problems. Local complications were mild and more frequent in patients "over" 65 years (13% vs 6%, in "under" 65), but recurrent hernias make the difference; a suction drainage can reduce the problems caused by a fluid collection in the inguinoscrotal area. Wantz technique reduces the operative time and, according to personal opinion, contributed to lower ischemic testicular complications to zero.


Amelioration of cancer stem cells in macrophage colony stimulating factor-expressing U87MG-human glioblastoma upon 5-fluorouracil therapy.
Macrophage colony stimulating factor (MCSF) regulates growth, proliferation and differentiation of haematopoietic cell lineages. Many cancers are known to secrete high level of MCSF, which recruit macrophages into the tumour micro-environment, supporting tumour growth. Herein, we report the cloning of MCSF and subsequent generation of U87MG expressing MCSF stable cell line (U87-MCSF). Cytotoxicity of anti-cancer drug 5-fluorouracil (5-FU) was evaluated on both U87MG and U87-MCSF cells. Interestingly, the proliferation of U87-MCSF cells was less (p<0.001) than that of U87MG cells alone, after treatment with 5-FU. Significant decrease in expression levels of cyclin E and A2 quantified by real time PCR analysis corroborated the reduced proliferation of 5-FU treated U87-MCSF cells. However, JC-1 staining did not reveal any apoptosis upon 5-FU treatment. Notch-1 upregulation induced a possible epithelial-mesenchymal transition in U87-MCSF cells, which accounted for an increase in the proportion of CD24(high)/CD44(less) cancer stem cells in U87-MCSF cells after 5-FU treatment. The elevated resistance of U87-MCSF cells towards 5-FU was due to the increase in the expressions (10.2 and 6 fold) of ABCB1 and mdm2, respectively. Furthermore, increase in expressions of ABCG1, mdm2 and CD24 was also observed in U87MG cells after prolonged incubation with 5-FU. Our studies provided mechanistic insights into drug resistance of U87MG cells and also described the pivotal role played by MCSF in augmenting the resistance of U87MG cells to 5-FU.


The risk of developing Creutzfeldt-Jakob disease in subjects with the PRNP gene codon 200 point mutation.
We determined the penetrance of the PRNP 200Lys mutation in the large cluster of Creutzfeldt-Jakob disease (CJD) cases among Jews of Libyan-Tunisian origin living in Israel, utilizing data from 52 carriers with definite or probable CJD and 34 unaffected mutation carriers. A life table analysis was carried out with development of CJD as the end point. The probability of developing CJD rose with age, fitting a second-order regression curve (R = 0.97, p < 0.001). The cumulative penetrance reached 50% at the age of 60 and 80% at 80 years. Including seven elderly possible CJD patients in the analysis made the penetrance approach 100% by age eighty. The penetrance of the mutation is high, and although age is a predominant influencing factor, other factors, such as gender, may also play a role.


Small nucleolar RNAs in plasma extracellular vesicles and their discriminatory power as diagnostic biomarkers of Alzheimer's disease.
The clinical diagnosis of Alzheimer's disease, at its early stage, remains a difficult task. Advanced imaging technologies and laboratory assays to detect Aβ peptides Aβ42 and Aβ40, total and phosphorylated tau in CSF provide a set of biomarkers of developing AD brain pathology and facilitate the diagnostic process. The search for biofluid biomarkers, other than in CSF, and the development of biomarker assays have accelerated significantly and now represent the fastest-growing field in AD research. The goal of this study was to determine the differential enrichment of noncoding RNAs (ncRNAs) in plasma-derived extracellular vesicles (EV) of AD patients and Cognitively Normal controls (NC). Using RNA-seq, we profiled four significant classes of ncRNAs: miRNAs, snoRNAs, tRNAs, and piRNAs. We report a significant enrichment of SNORDs - a group of snoRNAs, in AD samples compared to NC. To verify the differential enrichment of two clusters of SNORDs - SNORD115 and SNORD116, localized on human chromosome 15q11-q13, we used plasma samples of an independent group of AD patients and NC. We applied ddPCR technique and identified SNORD115 and SNORD116 with a high discriminatory power to differentiate AD samples from NC. The results of our study present evidence that AD is associated with changes in the enrichment of SNORDs, transcribed from imprinted genomic loci, in plasma EV and provide a rationale to further explore the validity of those SNORDs as plasma biomarkers of AD.


[Prednisolone therapy for Kawasaki disease].
Prednisolone therapy for acute phase Kawasaki disease had been considered to be controversial for several decades. However, recent clinical studies revealed that prednisolone therapy might improve coronary and clinical outcomes. The present paper reviews the roles of prednisolone therapy for acute phase Kawasaki disease patients.


[Cardiovascular and hemodynamic effects of derivatives of metoclopramide].
Two salicylamides, thiapride and sultopride, were found to provoke a dose-related depressor effect in the anesthetized dog. This effect was qualitatively similar to that of procaïnamide and metoclopramide. The hypotensive and negative inotropic effects were not related to the parasympathic system. In fact these compounds are weak autonomic antagonists. They do not exert any anticholinesterase activity. Intra-arterial administration provoked vasodilation.


Endovascular occlusion of pial arteriovenous macrofistulae, using pCANvas1 and adenosine-induced asystole to control nBCA injection.
Background In large-caliber pial macrofistulae (pMF), the combination of high blood flow velocity and large efferent artery diameter makes control over the endovascular vessel occlusion difficult and may result in the inadvertent venous passage of occlusive devices or embolic agents. Case descriptions Patient 1: A 27-year-old man presented with headache and ataxia. An infratentorial pMF supplied by both superior cerebellar arteries with venous ectasia was found. The first treatment attempt using balloons and coils failed since the position of either device could not be controlled because of a distal diameter of the feeding artery of 8 mm. In a second session a pCANvas1 (phenox) was deployed at the level of the arteriovenous connection and adenosine-induced asystole allowed the controlled injection of nBCA/Lipiodol with partial occlusion of the pMF. A remaining arteriovenous shunt was occluded under asystole in a third session. The procedures were well tolerated, the patient returned to normal and DSA confirmed the occlusion of the fistula. Patient 2: A 13-year-old boy with hereditary hemorrhagic teleangiectasia presented with an intracerebral hemorrhage from an aneurysm of the left MCA. Twelve weeks after the aneurysm treatment a feeding MCA branch (diameter 4.5 mm) of a right frontal pMF was catheterized. The macrofistula was occluded by deployment of a pCANvas1, followed by the injection of nBCAl/Lipiodol under adenosine-induced asystole. Conclusion pCANvas1 and adenosine-induced asystole allow a controlled injection of nBCA/Lipiodol for the endovascular occlusion of high-flow pMF without venous passage of the embolic agent.


Implantation of a dual-chamber pacemaker in a patient with dextrocardia and sick sinus syndrome: a case report.
Dextrocardia is a congenital abnormal position of the heart in which the main part of the heart is in the right chest, and the long axis of the heart points to the lower right. Cases of a combination of dextrocardia and sick sinus syndrome are rare. A 65-year-old female patient was admitted to hospital with palpitations and dizziness for 1 week. Mirror-image dextrocardia and sick sinus syndrome were diagnosed by an electrocardiogram, echocardiography, Holter monitoring, and X-rays. Finally, we successfully implanted a dual-chamber pacemaker into the patient. The patient had an uneventful recovery and was discharged when her symptoms had greatly improved 1 week later. When dextrocardia is present, using active fixation leads in the atrial and ventricular leads is easier for finding the pacing position with optimal sensing and pacing thresholds, and they reduce the incidence of falling off.


Antibody-producing cell responses to an isolated outer membrane protein and to complexes of this antigen with lipopolysaccharide or with vesicles of phospholipids from Proteus mirabilis.
Antibody-producing cell responses of mice to a protein isolated from the outer membrane of Proteus mirabilis were typical of the responses to a thymus-dependent antigen. The immunoglobulin G antibody-producing cell responses to the protein were increased after administration of the antigen complexed with either lipopolysaccharide or with vesicles of phospholipids extracted from P. mirabilis. The protein in turn significantly increased the immune response to lipopolysaccharide and also converted this response from predominantly immunoglobulin M to predominantly immunoglobulin G.


Promoter specificity and deletion analysis of three heat stress transcription factors of tomato.
Transient expression assays in transformed tobacco (Nicotiana plumbaginifolia) mesophyll protoplasts were used to test the activity of three tomato heat stress transcription factors, HSF24, HSF8 and HSF30, in a trans-activation and a trans-repression assay. The results document differences between the three HSFs with respect to their response to the configuration of heat stress promoter elements (HSEs) in the reporter construct (promoter specificity) and to the stress regime used for activation. Analysis of C-terminal deletions identified acidic sequence elements with a central tryptophan residue, which are important for HSF activity control. Surprisingly, heterologous HSFs from Drosophila and human cells, but not from yeast, were also functional as heat stress-induced transcription factors in this tobacco protoplast system.


Fatal snakebite in Australia.
More than 20 venomous snakes found in Australia belong to the family Elapidae . Although their venom delivery system is less efficient than that of the Viperidae , their venoms are extremely potent. The active components of Australian elapid venoms include neurotoxins, myotoxins , hemolysins, and factors producing hypocoagulability. The groups at particular risk of snakebite are children, agricultural workers, bushwalkers , and herpetologists . A high index of suspicion must be maintained when investigating cases of sudden unexpected death in these groups. The bite wound itself may be particularly easily overlooked as there is little local reaction to many elapid bites. The necropsy findings in fatalities are predictable from the known properties of the venoms of the various species. However, these findings are non-specific and the diagnosis ultimately rests upon the demonstration of the presence of venom by immunoassay. For this purpose swabs from the bite site, the overlying clothing, the excised bite site, the local and regional lymph nodes, urine, and blood may be used.


The Breakthroughs in Cancer Immune Checkpoint Based Therapy: A Review of Development in Immune Checkpoint Study and its Application.
Recently, immunotherapy has attracted more attentions to fight cancer due to its selectivity, long lasting effects, and demonstrated better overall survival and tolerance, when compared to patients treated with conventional chemotherapy or radiotherapy alone. The anti-tumor response of patient with cancer is improved either by increasing the effector cell number, the production of soluble mediators, or by modulating the host's immune checkpoint. Over the last decades, many new approaches in immunotherapy have been developed, such as immune checkpoint inhibitors, chimeric antigen receptor T cells (CART), specific T-cell -receptor T cells (TCRT) and cancer vaccine, some of which has been approved by FDA to treat several cancer types. Among them, immune checkpoint based therapy has shown tremendous promise in both solid and hematological malignancies, and has significantly improved overall survival in patients with advance cancer. Many immuno checkpoints has been discovered and demonstrated. However, there is still lack of a complete summary for them. Thus, the present review focuses on immune checkpoint study and their latest application.


On Eulerian versus Lagrangian models of mechanical blood damage and the linearized damage function.
Two limitations have been discovered in the derivation of the Eulerian method of hemolysis prediction using a linearized blood damage function. First is that in the transformation from the Lagrangian material volume of the original power-law model to a fixed Eulerian control volume, the spatial dependence of duration of exposure to fluid stress was neglected. This omission has the implication that the Eulerian method as reported is valid only for steady, uniaxial flow in which velocity is constant along streamlines. The second issue is related to linearization, which involves distributing an exponent across an integral. This operation is valid only for limited conditions that include the exponent being unity (which is not the case for any power-law hemolysis models) or the blood damage function being constant throughout the flow regime. These constraints severely restrict the applicability of the Eulerian method. An example problem is presented that demonstrates that the source term of the Eulerian method as reported does not account for differences in velocity between 2 similar flows. Correcting the source term to match the hemolysis prediction to that of the original, unlinearized method requires an analytical description of the flow field that may not be easily obtained for the complex flows in some cardiovascular devices.


Catatonic Schizophrenia: Cases with Possible Genetic Predisposition.
Catatonic schizophrenia is defined by catatonia seen either with alternating phases of stupor and motor rigidity or the extreme phase of catatonic excitement. This variant of schizophrenia has been identified with poor prognosis, mainly due to the higher association with negative symptoms and young age onset. In this paper, we illustrate a similar clinical picture of catatonic schizophrenia in two brothers, with no genetic predisposition to schizophrenia and no proximal stressors apart from the aggressive/violent behavior of their elder brother. Case presentation 1 (Patient A): An 18-year-old male from a lower socio-economic class with no previous mental health issues presented to the emergency department with complete mutism, marked psychomotor retardation, posturing along with a refusal to drink or eat, and complete lack of self-care for about two months. The diagnosis of catatonic schizophrenia was made, and the patient was started orally on aripiprazole and lorazepam. On the third day of admission, noticeable changes were observed, and in the following days, he started eating and going to the toilet while still being completely mute. After two weeks on treatment, he started responding with one-word answers. Case presentation 2 (Patient B): The biological brother of patient A, a 30-year-old male, presented on the same day with an identical history of mutism, decreased psychomotor activity, posturing along with a refusal to drink or eat, and lack of self-care for the past few months. The diagnosis of catatonic schizophrenia was made. The patient was started orally on both, olanzapine and lorazepam. He showed a quicker response to treatment with the maintenance of eye contact on the second day of treatment and started giving short answers to questions on the fifth post-admission day. We here discuss a possible genetic predisposition to catatonic schizophrenia and its initial improvement with lorazepam and subsequent treatment with olanzapine proving to be more efficacious than aripiprazole.


Temporary Remedy.
The nationwide shortage of doctors shapes the health care landscape in many ways, mostly through staffing headaches and gaps in care. But the shortage has been a blessing for at least one group of health care practitioners - locum tenens physicians. More and more, locum tenens doctors are taking the places of permanent physicians.


Histopathological features of L-asparaginase-induced liver disease.
We studied the histopathological changes of liver in four patients who developed hepatomegaly and abnormal liver chemistry tests 2 to 20 days following administration of L-asparaginase as a part of a combination chemotherapy regimen for treatment of acute lymphoblastic leukemia. The severity of the liver disease due to L-asparaginase was unpredictable. One patient developed acute fulminant hepatic failure and required liver transplantation. The most consistent pathological change, observed in all four cases, was diffuse steatosis. Other changes included patchy hepatocyte necrosis, mixed inflammatory cell infiltrates in the portal tracts, and variable degrees of hepatocellular, or canalicular cholestasis, or a combination of these.


Predicting the effective temperature of a glass.
We explain the findings by Di Leonardo et al. [Phys. Rev. Lett. 84, 6054 (2000)10.1103/PhysRevLett.84.6054] that the effective temperature of a Lennard-Jones glass depends only on the final density in the volume and/or temperature jump that produces the glass. This is not only a property of the Lennard-Jones liquid, but a feature of all strongly correlating liquids. For such liquids data from a single quench simulation provide enough information to predict the effective temperature of any glass produced by jumping from an equilibrium state. This prediction is validated by simulations of the Kob-Andersen binary Lennard-Jones liquid and shown not to apply for the nonstrongly correlating monatomic Lennard-Jones Gaussian liquid.


Defining Peer-to-Peer Accountability From the Nurse's Perspective.
The aim of this study was to define and create a conceptual model for peer-to-peer accountability (P to PA). Many organizations cite the importance of peer accountability (PA) as essential in ensuring patient safety. Professionalism in nursing requires self-regulation of practice and PA. Although discussed in the literature, P to PA is not conceptually defined. A grounded theory study design with constant comparative data collection and analysis was used to explore nurses' definitions of P to PA and their perceptions of motivators and barriers to engaging in P to PA. Transcripts of digital recordings of all interviews were analyzed using line-by-line coding until identified themes emerged. P to PA was defined as the act of speaking up when one observes a peer not practicing to acceptable standards. A conceptual model illustrates the antecedents, attributes, and consequences of P to PA. P to PA is the professional responsibility of every nurse and healthcare provider and is essential for safe patient care. The conceptual definition facilitates actualization of P to PA in practice.


[Quality of some working places in Milan].
Questionnaires were distributed to the responsibles and to the dependents of structures with high turnover of people, to have informations about numbers and typology of instruments, habits of dependents and type of cleanings. In these structures, microbiologic quality of air and of informatic instruments' surfaces were evaluated and efficacy of treatment with some products of cleaning and sanification was verified. Air microbiological contamination was comparable to that found in other similar researchs. Fecal contamination indicators were not found on instruments' surfaces, both before, and after treatment; in 8 cases (3%) Staphylococcus aureus was isolated and in 6 cases (2%) species of not pathogenous staphylococci were isolated. At the beginning of the working day, mean values of total aerobic bacterial count at 22 degrees and 37 degrees, were low, settled to zero after treatment and increased progressively during the week. Infective risk for workers can be considered insignificant because of low microbiological contamination. Specific sanification products seem not to be necessary, since common products of cleaning have the same efficacy.


Publisher's Note: Evidence for Exotic Hadron Contributions to Λ_{b}^{0}→J/ψpπ^{-} Decays [Phys. Rev. Lett. 117, 082003 (2016)].
This corrects the article DOI: 10.1103/PhysRevLett.117.082003.


Titanium versus stainless steel for anterior spinal fusions: an analysis of rod stress as a predictor of rod breakage during physiologic loading in a bovine model.
In vitro biomechanical evaluation of rod stress during physiologic loading of anterior scoliosis instrumentation. To determine effects of material properties and rod diameter on rod stresses in anterior scoliosis instrumentation. Relationships between instrumentation dimensions, materials, and potential rod failure in anterior scoliosis instrumentation remain unclear. Eighteen immature bovine spines were randomized to 3 groups: 1) 4.0-mm stainless steel, 2) 5.0-mm stainless steel, and 3) 4.75-mm titanium alloy. Spines underwent physiologic tests in flexion-extension, lateral bending, and torsion. Rod surface strains were converted to rod stress and normalized to each material's yield stress. Construct stiffness and the normalized rod stresses were compared with a one-way ANOVA (P < 0.05). The 4.0-mm steel and 4.75-mm titanium construct stiffness was similar across all tests. The 5.0-mm steel system was significantly stiffer than 4.0-mm steel (lateral bending/torsion) and 4.75-mm titanium (torsion/flexion) constructs. Rod surface stress was significantly lower for the 4.75-mm titanium rod compared with 4.0-mm and 5.0-mm steel rods for all tests. The percentage of yield stress was lowest for the 4.75-mm Ti rod for all tests due to titanium's greater yield stress. This suggests the 4.75-mm rod has a lower fatigue failure risk than either steel construct.


Endovascular management of an isolated dissection of the celiac artery following a blunt deceleration-type trauma.
A 29-year-old, otherwise healthy male was brought to the emergency department after suffering an injury from falling from the roof of his house. Whole-body computed tomography (CT) showed a small ostial dissection of the celiac artery (CA) associated with a pseudoaneurysm, thrombosis, and retroperitoneal periaortic hematoma. The celiac trunk was otherwise normal with good distal patency and no signs of intestinal ischemia. The patient was successfully managed via an endovascular approach with the placement of a covered stent in the CA.


[Primary aldosteronism: new insights into familial forms].
Primary aldosteronism (PA) is characterized by autonomic aldosterone production, usually leading to severe hypertension and hypokalaemia. PA is a heterogeneous condition caused by sporadic adrenal adenoma, bilateral adrenal hyperplasia or rare familial forms. Familial aldosteronism type 1 is caused by a hybrid gene that codes for an ACTH-sensitive form of aldosterone synthase. Familial aldosteronism type 3 was recently recognized as a new form of PA caused by mutation in KCNJ5. The clinical manifestations vary from life-threatening PA and pronounced adrenal hyperplasia to milder forms. In addition to germline mutations in KCNJ5, somatic KCNJ5 mutations are present in about 40% of aldosterone-producing adrenal adenomas. Mutations in three other genes are also regularly observed. All these mutations cause increased aldosterone synthase activity, eventually leading to PA. In patients under 20 with PA, familial forms must be excluded before proceeding to adrenalectomy.


Colorectal adenoma and carcinoma specific miRNA profiles in biopsy and their expression in plasma specimens.
MiRNA expression markers are well characterized in colorectal cancer (CRC), but less is known about miRNA expression profiles in colorectal adenomas. Genome-wide miRNA and mRNA expression analyses were conducted through the colorectal adenoma dysplasia sequence. Furthermore, analysis of the expression levels of miRNAs in matched plasma samples was performed, focusing on biomarker candidates; miRNA and mRNA expression analyses were performed on colorectal biopsies and plasma samples (20 normals; 11 tubular and 9 tubulovillous adenomas; 20 colorectal carcinomas) by miRNA 3.0 and Human Transcriptome Array (Affymetrix) and validated by RT-qPCR. Microarray data were analyzed using Expression Console and mRNA targets were predicted using miRWALK 2.0. Based on microarray analysis, 447 miRNAs were expressed in tissue and 320 in plasma. Twelve were upregulated (miR-31, 8-fold p < 0.001) and 11 were downregulated (miR-10b 3-fold p < 0.001) in neoplastic lesions compared to normal group. Eleven miRNAs showed altered expression between adenoma subtypes (miR-183 2.8-fold change, p < 0.007). Expression level of 24 miRNAs differed between adenoma and CRC groups (including miR-196a, 3.5-fold). Three miRNAs (miR-31, miR-4506, miR-452*) were differentially expressed in adenoma compared to normal both in tissue and plasma samples. miRNA expression data were confirmed by RT-PCR both in plasma and matched tissue samples. MiRNAs showed characteristic expression changes during CRC development in tissue. miRNAs were also presented in plasma and positively correlated with matched tissue expression levels. The identified miRNA expression changes could be verified RT-PCR methods facilitating routine application.


Measurement and modeling of pulsed microchannel plate operation (invited).
Microchannel plates (MCPs) are a standard detector for fast-framing x-ray imaging and spectroscopy of high-temperature plasmas. The MCP is coated with conductive striplines that carry short duration voltage pulses to control the timing and amplitude of the signal gain. This gain depends on the voltage to a large exponent so that small reflections or impedance losses along the striplines can have a significant impact on the position-dependent amplitude and pulse width of the gain. Understanding the pulsed gain response therefore requires careful measurements of the position- and time-dependent surface voltage coupled with detailed modeling of the resulting electron cascade. We present measurements and modeling of the time- and space-dependent gain response of MCP detectors designed for use at Sandia National Laboratories' Z facility. The pulsed gain response is understood through measurements using a high impedence probe to determine the voltage pulse propagating along the stripline surface. Coupling the surface voltage measurements with Monte Carlo calculations of the electron cascade in the MCP provides a prediction of the time- and position-dependent gain that agrees with measurements made on a subpicosecond UV laser source to within the 25% uncertainty in the simulations.


Substantial increase in listeriosis, Denmark 2009.
In 2009, 97 cases of listeriosis were reported in Denmark (1.8 per 100,000), a significant rise over the previous year. The increase was seen both in cases of bacteraemia and meningitis and affected mainly people aged 70 years and older. A foodborne outbreak of eight cases was identified by pulsed-field gel electrophoresis typing. No explanation has so far been found for the marked increase in incidence. An increasing trend has been observed since 2003 and possible explanations are discussed.


Differential regulation and function of the Fas/Fas ligand system in human trophoblast cells.
Trophoblast rejection, which is characterized by increased apoptosis, is mediated by T helper (Th)-1, or proinflammatory, cytokines, whereas Th-2, or anti-inflammatory, cytokines confer immune protection and facilitate implantation. We investigated the role of both types of cytokines on the expression and function of the Fas/Fas ligand (FasL) apoptotic pathway in trophoblast cells. First-trimester human trophoblast primary-culture cells as well as A3 and HTR/8 trophoblast cell lines were treated with proinflammatory cytokines such as interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNFalpha) and with the anti-inflammatory cytokines interleukin (IL)-6 and IL-10. Sensitivity to Fas-mediated apoptosis was measured using an activating anti-Fas monoclonal antibody. Cell viability was evaluated using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) and CellTiter 96 assay. Fas/FasL mRNA and protein expression levels were determined using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Trophoblast cells normally express FasL, but low levels of Fas, and they are resistant to Fas-mediated apoptosis. IFN-gamma and TNFalpha promote Fas expression and sensitivity, whereas IL-6 and IL-10 increase the resistance of trophoblast cells to Fas-mediated apoptosis. Furthermore, IL-10 treatment activates FLICE-like inhibitory protein (FLIP), a downstream inhibitor of Fas apoptotic signaling. Although trophoblast cells express Fas, susceptibility to Fas does not necessarily correlate with its expression. In this study, we demonstrate that Th-2 cytokines increase the resistance of trophoblast cells to Fas-mediated apoptosis either by inhibiting Fas expression or by inducing FLIP activation. This "trophoblast-cytokine-Fas/FasL triad" determines the ability of the Fas/FasL system to regulate trophoblast viability and, consequently, the success or failure of pregnancy.


The endoplasmic reticulum (ER) translocon can differentiate between hydrophobic sequences allowing signals for glycosylphosphatidylinositol anchor addition to be fully translocated into the ER lumen.
The signal sequence within polypeptide chains that designates whether a protein is to be anchored to the membrane by a glycosylphosphatidylinositol (GPI) anchor is characterized by a carboxyl-terminal hydrophobic domain preceded by a short hydrophilic spacer linked to the GPI anchor attachment (omega) site. The hydrophobic domain within the GPI anchor signal sequence is very similar to a transmembrane domain within a stop transfer sequence. To investigate whether the GPI anchor signal sequence is translocated across or integrated into the endoplasmic reticulum membrane we studied the translocation, GPI anchor addition, and glycosylation of different variants of a model GPI-anchored protein. Our results unequivocally demonstrated that the hydrophobic domain within a GPI signal cannot act as a transmembrane domain and is fully translocated even when followed by an authentic charged cytosolic tail sequence. However, a single amino acid change within the hydrophobic domain of the GPI-signal converts it into a transmembrane domain that is fully integrated into the endoplasmic reticulum membrane. These results demonstrated that the translocation machinery can recognize and differentiate subtle changes in hydrophobic sequence allowing either full translocation or membrane integration.


To use or not to use continuous passive motion post-total knee arthroplasty presenting functional assessment results in early recovery.
Continuous passive motion (CPM), though of doubtful value, is yet routinely practiced post-total knee arthroplasty (TKA). We prospectively distributed 84 patients with TKA to 1 of the 3 standard rehabilitation regimes: no-CPM, 1-day-CPM, and 3-day-CPM. We recorded a unique "Timed up and go" test besides pain, Western Ontario and McMaster Universities (WOMAC), short form-12 (SF-12), range of motion, knee and calf swelling, and wound healing parameters. Our standardized and elaborate measurements preoperatively and on postoperative days 3, 5, 14, 42, and 90 showed no statistically significant difference among the 3 groups in each parameter. We concluded that CPM gives no benefit in immediate functional recovery post-TKA, and in fact, the postoperative knee swelling persisted longer. We have since then discontinued its use in our patients without any untoward effect.


Controlled study of exclusion of dietary vasoactive amines in migraine.
To assess the effects of dietary vasoactive amines in the aetiology of childhood migraine, 39 children were randomly allocated to either a high fibre diet low in these substances or a high fibre diet alone. Both groups of children showed a significant decrease in the number of headaches and there was no significant difference between the two groups. Dietary vasoactive amines have not been shown in this study to influence childhood migraine. The improvement seen in both groups emphasises the need for a control diet in studies designed to show that dietary manipulation improves disease.


Normal and cancerous mammary stem cells evade interferon-induced constraint through the miR-199a-LCOR axis.
Tumour-initiating cells, or cancer stem cells (CSCs), possess stem-cell-like properties observed in normal adult tissue stem cells. Normal and cancerous stem cells may therefore share regulatory mechanisms for maintaining self-renewing capacity and resisting differentiation elicited by cell-intrinsic or microenvironmental cues. Here, we show that miR-199a promotes stem cell properties in mammary stem cells and breast CSCs by directly repressing nuclear receptor corepressor LCOR, which primes interferon (IFN) responses. Elevated miR-199a expression in stem-cell-enriched populations protects normal and malignant stem-like cells from differentiation and senescence induced by IFNs that are produced by epithelial and immune cells in the mammary gland. Importantly, the miR-199a-LCOR-IFN axis is activated in poorly differentiated ER- breast tumours, functionally promotes tumour initiation and metastasis, and is associated with poor clinical outcome. Our study therefore reveals a common mechanism shared by normal and malignant stem cells to protect them from suppressive immune cytokine signalling.


Issues of testing for hGH: collection.
Urine has limitations as a substrate for anti-doping efforts and within the last decade sports organizations have begun collecting blood samples in cooperation with anti-doping laboratories. Within the US, authorization to perform a blood draw is determined by the scope-of-practice licensure regulations in each state. Blood collection requires equipment not necessary in standard anti-doping urine collections. There is a need for greater privacy and cleanliness than the usual site for urine collection provides. One of the most challenging aspects of a blood collection is complying with the guidelines for specimen transport. In addition to the standard chain-of-custody protocols that collectors must follow for urine, blood samples call for additional handling to ensure they arrive at the anti-doping laboratory in a condition that allows for analysis. While the invasiveness of the process increases liability risk to the collector, following established standards, guidelines and anti-doping protocols will minimize risk. One significant advantage of blood over urine in anti-doping settings is that specimen adulteration or manipulation is nearly impossible. Therefore, for blood-only collections, sports organizations should explore opportunities for collaboration with the American Red Cross or private blood-collection centres/clinics.


Precision of Taylor Dispersion.
Taylor dispersion is capable of measuring accurately the hydrodynamic radius over several orders of magnitude. Accordingly, it is now a highly competitive technique dedicated to characterizing small molecules, proteins, macromolecules, nanoparticles, and their self-assembly. Regardless, an in-depth analysis addressing the precision of the technique, being a key indicator of reproducibility, is not available. Benefiting from analytical modeling and statistical analysis, we address error propagation and present a comprehensive theoretical study of the precision of Taylor dispersion. Theory is then compared against experiment, and we find full consistency. Our results are most helpful when the design, objectives, or control of analytical quality is in focus.


Determination of the Cell Permissiveness Spectrum, Mode of RNA Replication, and RNA-Protein Interaction of Zika Virus.
Two lineages of Zika virus (ZIKV) have been classified according to the phylogenetic analysis: African and Asian lineages. It is unclear whether differences exist between the two strains in host cell permissiveness, this information is important for understanding viral pathogenesis and designing anti-viral strategies. In the present study, we comparatively studied the permissive spectrum of human cells for both the African (MR766) and Asian strains (PRVABC59) using an RNA in situ hybridization (RISH) to visualize RNA replication, an immunofluorescence technology, and a western blot assay to determine viral protein production, and a real-time RT-PCR to examine viral RNA multiplication level. The experiments were undertaken in the condition of cell culture. We identified several human cell lines, including fibroblast, epithelial cells, brain cells, stem cells, and blood cells that are susceptible for the infection of both Asian and African strains. We did not find any differences between the MR766 and the PRVABC59 in the permissiveness, infection rate, and replication modes. Inconsistent to a previous report (Hamel et al. JVI 89:8880-8896, 2015), using RISH or real-time RT-PCR, we found that human foreskin fibroblast cells were not permissive for ZIKV infection. Instead, human lung fibroblast cells (MRC-5) were fully permissive for ZIKV infection. Surprisingly, a direct interaction of ZIKV RNA with envelop (E) protein (a structure protein) was demonstrated by an RNA chromatin immunoprecipitation (ChIP) assay. Three binding sites were identified in the ZIKV RNA genome for the interaction with the E protein. Our results imply that the E protein may be important for viral RNA replication, and provide not only the information of ZIKV permissiveness that guides the usage of human cells for the ZIKV studies, but also the insight into the viral RNA-E protein interaction that may be targeted for intervention by designing small molecule drugs.


[Cemented femoral prosthesis with calcar replacement for intertrochanteric hip fractures in the elderly].
The increase in hip fractures among the elderly and the presence of intercurrent diseases makes it necessary to institute treatments that help patients resume their activities as soon as possible. Analyze the clinical course of patients with a cemented femoral endoprosthesis with calcar replacement and determine the time to resuming walking. 18 patients were seen from January 2003 to December 2006, all of them with a diagnosis of Tronzo T IV and V intertrochanteric hip fractures, over 70 years of age and treated primarily with a cemented femoral endoprosthesis with calcar replacement. The total number of patients was 18, 14 females (77.77%) and 4 males (22.22%). The fracture types were: Tronzo grade IV in 11 patients and grade V in 7 patients; mean age was 78 years and their functional capacity in the immediate postoperative period was: onset of walking with full weightbearing at 48 hours in 100% of patients. Return to usual activities occurred at 4 weeks in 17 patients (94.44%). Due to the complexity of these fractures and the patients' bone quality, there are limited chances to treat them with osteosynthesis, as the latter would increase patients' morbidity and mortality. Patients could undergo early rehabilitation.


[Corrigendum] Genome‑wide DNA methylation analysis of uterosacral ligaments in women with pelvic organ prolapse.
Following the publication of this paper, the authors have realized that the authors' address affiliation was not presented as it should have officially appeared. The address affiliation should have appeared as "Department of Gynecology" (not "Gynecology Department"), Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing. Therefore, the authors' affiliation address should have been written as follows: Lifang Zhang, Ping Zheng, Aihong Duan, Yan Hao, Chang Lu and Dan Lu. Department of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, P.R. China. The authors confirm that there are no further errors in the study, and all the authors agree to this correction. The authors regret this error in the affiliation, and apologize for any inconvenience caused. [the original article was published in the Molecular Medicine Reports 19: 391‑399, 2019; DOI: 10.3892/mmr.2018.9656].


Cat scratch disease presenting with peripheral facial nerve paralysis.
Acquired peripheral facial nerve paralysis is a relatively common disorder that affects both children and adults. The most frequent nontrauma-related etiologies in otherwise neurologically intact patients are idiopathic (Bell's palsy) and infectious, which includes otitis media, herpes zoster, Lyme disease, herpes simplex virus, Epstein-Barr virus, and Mycoplasma pneumoniae. Cat scratch disease (CSD) is typically a subacute, regional lymphadenitis caused by Bartonella henselae that is seen in children and young adults. CSD most often has a benign, self-limited course. However, 11% of CSD patients may present atypically, most commonly with Perinaud's oculoglandular syndrome or acute encephalopathy. We present a child with the first reported case of acute facial nerve paralysis in serologically proven CSD with typical lymphadenitis.


The metalloproteinase ADAM10 requires its activity to sustain surface expression.
The metalloproteinase ADAM10 critically contributes to development, inflammation, and cancer and can be controlled by endogenous or synthetic inhibitors. Here, we demonstrate for the first time that loss of proteolytic activity of ADAM10 by either inhibition or loss of function mutations induces removal of the protease from the cell surface and the whole cell. This process is temperature dependent, restricted to mature ADAM10, and associated with an increased internalization, lysosomal degradation, and release of mature ADAM10 in extracellular vesicles. Recovery from this depletion requires de novo synthesis. Functionally, this is reflected by loss and recovery of ADAM10 substrate shedding. Finally, ADAM10 inhibition in mice reduces systemic ADAM10 levels in different tissues. Thus, ADAM10 activity is critically required for its surface expression in vitro and in vivo. These findings are crucial for development of therapeutic ADAM10 inhibition strategies and may showcase a novel, physiologically relevant mechanism of protease removal due to activity loss.


Paravertebral mass in a patient with hemolytic anemia: computed tomographic findings.
Extramedullary hematopoiesis is characterized by the presence of hematopoietic tissue outside of the bone marrow and is typically associated with chronic hemolytic anemias. Intrathoracic extramedullary hematopoiesis is a rare and usually asymptomatic condition. The authors report a case of a 57-year-old man with intrathoracic extramedullary hematopoiesis and hereditary spherocytosis. Clinical and laboratory evaluation, together with radiological findings, are described. The diagnosis of the disease was confirmed by tissue biopsy.


Toxicity of 1,1,1-trichloro-2-propanone in Sprague-Dawley rats.
1,1,1-Trichloro-2-propanone (1,1,1-TCP) has been identified as a chlorination by-product in finished drinking water supplies. Since little was known of its oral toxicity, exposure studies were conducted with male and female Sprague-Dawley rats (10 males and 10 females/group) exposed by corn oil gavage at 0, 16, 48, 161, or 483 mg/kg for 10 d or 0, 30, 90, or 270 mg/kg for 90 d. Evaluations included mortality, clinical signs, body weight, food consumption, ophthalmology, hematology, clinical chemistry, urinalysis, organ weights, gross pathology, and histopathology. In the 10-d study, severe toxicity was observed at the highest dose level, since most treated animals (8/10 males and 7/10 females) died. Toxicity was also noted at 161 and 48 mg/kg. At 161 mg/kg, 2 males died and an increase in liver weights in both sexes was observed. Acanthosis and hyperkeratosis of the forestomach was present in males and females at 48 mg/kg and above. In the 90-d study, toxicity was significant at 270 mg/kg, with acanthosis and hyperkeratosis of the forestomach evident in most animals and ataxia in about one-half of them. Retinal degeneration, increased serum potassium, and increased blood urea nitrogen were present in females and increased blood calcium in males at that same dose level. Acanthosis and hyperkeratosis were observed in both sexes, and retinal degeneration was prominent in 2 females at 90 mg/kg. It was concluded that 16 mg/kg was the NOAEL (no observed adverse effect level) for the 10-d study while 30 mg/kg was the NOAEL for the 90-d exposure of Sprague-Dawley rats to 1,1,1,-trichloro-2-propanone.


Activation of T lymphocytes by cross-linking of glycophospholipid-anchored Thy-1 mobilizes separate pools of intracellular second messengers to those induced by the antigen-receptor/CD3 complex.
T cells can be activated, not only by the conventional (antigen-receptor/CD3 complex) route, but also by cross-linking any one of their lipid-anchored surface glycoproteins. We have compared early transmembrane signalling events mediated through CD3 with those mediated through Thy-1, a lipid-linked surface glycoprotein, on the human lymphoid cell line Jurkat and transfectants expressing higher levels of Thy-1. Cross-linking of Thy-1 causes immediate phosphatidylinositol (PI) turnover and an influx of extracellular Ca2+, while releasing very little Ca2+ from intracellular stores. CD3 activation, on the other hand, causes PI turnover which releases intracellular Ca2+, and only secondarily induces an influx of extracellular ions. The Thy-1 response is detectable at very low levels of surface Thy-1, and is not mimicked by enzymatic removal of lipid-linked proteins from the cell surface. The Thy-1-induced Ca2+ influx is more sensitive to L channel blockers than the CD3-mediated flux. These results indicate that the initial stages of Thy-1-mediated activation involve the rapid and extensive mobilization of the intracellular second messengers, PI and Ca2+, by mechanisms separate to those activated by the antigen-receptor/CD3 complex.


Effect of pyrenzepine on gastric secretion in Shay rats.
The degranulation of the gastric mastocytes, gastric cAMP levels and intragastric pH were studied in Shay rats after the intraduodenal administration of 20 mg/kg pyrenzepine chlorhydrate. There was a significant increase in the intragastric pH after the administration of this substance, reaching its maximum alkalization 120 min later. The cAMP levels decreased significantly and reached their lowest levels 60 min later. The possible antihistaminic effect of the drug was also studied. Pyrenzepine did not reduce the acid secretion induced by the subcutaneous administration of different doses of histamine. The results show that the decrease in the cAMP gastric levels and in the acid secretion was not due to an antihistaminic effect of pyrenzepine chlorhydrate.


Effects of dexamethasone on proliferation of and fibronectin synthesis by human primary prostatic stromal cells in vitro.
Glucocorticoids are anti inflammatory stress hormones and have been suggested to be involved in a large number of pathological processes. To test the effects of glucocorticoids on stromal prostatic cell growth and proliferation in vitro, the influence of a synthetic glucocorticoid (dexamethasone, dex) on recently established human primary cells from prostatic stroma (hPCPs) was analysed. The localization and distribution of the glucocorticoid receptor (GR) was investigated by immunohistochemistry. In addition, expression of the active isoform of the receptor (alpha-GR) was examined by reverse transcription PCR, and the effect of different doses of dex on proliferation of the stromal cells evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and amido black assays. alpha-GR mRNA was expressed by the hPCPs, and the GR protein was detected in the cytoplasm and nucleus of these cells. Incubating the cells with dex resulted in an enhanced cell proliferation that was mainly restricted to the fibroblasts. Moreover, fibronectin (FN) gene expression and secretion of the protein was increased by high doses of dex (> or = 10(-8) M), whereas low doses of dex (10(-10)M) showed no effect. Human prostatic stromal cells show sensitivity to dex in vitro, resulting in an increase in cell proliferation and FN synthesis. The authors assume that locally accumulating glucocorticoids can also influence the regulation of cell growth and extracellular matrix synthesis in the human prostate in vivo and may play a role in the pathologically altered prostate.


Prolactin-releasing action of LRF: a central catecholamine mediated event?
Decline of plasma dopamine (DA), norepinephrine (NE), and epinephrine (EP) levels after iv administration of a 100 microgram bolus of LRF has been reported in normal men. This finding has been used to support the concept of a central dopamine mediated mechanism for LRF-induced PRL release. In the present study (including 5 postmenopausal women and 4 normal men), no detectable changes were found in plasma levels of DA, NE, EP and the DA metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC) following LRF (100 microgram iv) administration. These results pertain to both groups of subjects, although the PRL increment was 2-fold greater in the postmenopausal women than in the normal men. It is concluded that peripheral administration of LRF causes no acute decline of plasma catecholamines, and that the LRF-induced release of PRL may be based on the recent demonstration in vitro of a paracrine effect, gonadotrope to lactotrope.


Packing quality, protein binding capacity and separation efficiency of pre-packed columns ranging from 1 mL laboratory to 57 L industrial scale.
Pre-packed chromatography columns are routinely used in downstream process development and scale-down studies. In recent years they have also been widely adopted for large scale, cGMP manufacturing of biopharmaceuticals. Despite columns being qualified at their point of manufacture before release for sale, the suitability of pre-packed chromatography columns for protein separations at different scales has not yet been demonstrated. In this study, we demonstrated that the performance results obtained with small scale columns (0.5 cm diameter × 5 cm length, 1 mL column volume) are scalable to production sized columns (60 cm diameter × 20 cm length, 57 L column volume). The columns were characterized with acetone and blue dextran pulses to determine the packing density and packed bed consistency. Chromatography performance was evaluated with breakthrough curves including capacity measurements and with separation of a ternary protein mixture (lysozyme, cytochrome C and RNase A) with a step gradient. The equilibrium binding capacity and dynamic binding capacity were equivalent for all columns. The step gradient separation of the ternary protein mixture displayed similar peak profiles when normalized in respect to column volume and the eluted protein pools had the same purities for all scales. Scalable performance of pre-packed columns is demonstrated but as with conventionally packed columns the influence of extra column volume and system configurations, especially buffer mixing, must be taken into account when comparing separations at different scales.


Chaotic mixer using electro-osmosis at finite Péclet number.
Two pressure-driven streams of two miscible liquids can only mix by diffusion in microfluidic channels because of the low Reynolds number. We present an idea to generate mixing by "chaotic advection" in microscale geometries. That is, we consider using induced-charge electro-osmosis to generate a second flow and then modulate between the pressure-driven and induced-charge flows. By using the combined method consisting of the boundary element method, the Lagrangian particle tracking method, and the random-walk method, we analyze mixing efficiency, mixing time, and mixing length, with the effects of modulation frequency and molecular diffusivity, and compare our proposed mixer with other mixers. By this analysis, we find that chaotic mixing can be produced efficiently in a microfluidic channel by switching between pressure-driven and induced-charge flows in a wide range of Péclet number under the specific condition of Strouhal number. By using our proposed mixer, we can expect to realize efficient chaotic mixing with minimum voltage in an ordinary flow channel with a simple structure without an oblique electric field even at large Péclet number.


Cooperative photochemical reaction mechanism of femtosecond laser-induced photocoloration in spirooxazine microcrystals.
Microcrystalline powders of spirooxazine and spiropyran compounds do not show photocoloration under steady-state illumination, whereas they undergo photochromism on intense femtosecond laser-pulse excitation. We investigated the characteristic mechanism of the crystalline photochromism by studying the photocoloration of spironaphthooxazine (SNO) and its chloro-substituted derivative (Cl-SNO) with our femtosecond diffuse-reflectance spectroscopic system. In particular, femtosecond double-pulse excitation using 390+780-nm pulses and 390+390-nm pulses, with a variable time interval between the two pulses, was applied to reveal an intermediate species involved in the photocoloration. Although 780-nm excitation of an intermediate produced by 390-nm excitation did not lead to isomerization, the 390+390-nm excitation resulted in photocoloration. The yield for SNO decreased on increasing the interval from 40 ps to 5 ns, while that for Cl-SNO was constant. The photocoloration mechanism in the crystalline phase is considered from the viewpoint of the time-dependent density of short-lived transient species, and it is concluded that cooperative interactions of excited states and nonplanar open forms play an important role in femtosecond laser-induced photochromism in these crystals.


Molecular mechanisms for the conversion of zymogens to active proteolytic enzymes.
Proteolytic enzymes are synthesized as inactive precursors, or "zymogens," to prevent unwanted protein degradation, and to enable spatial and temporal regulation of proteolytic activity. Upon sorting or appropriate compartmentalization, zymogen conversion to the active enzyme typically involves limited proteolysis and removal of an "activation segment." The sizes of activation segments range from dipeptide units to independently folding domains comprising more than 100 residues. A common form of the activation segment is an N-terminal extension of the mature enzyme, or "prosegment," that sterically blocks the active site, and thereby prevents binding of substrates. In addition to their inhibitory role, prosegments are frequently important for the folding, stability, and/or intracellular sorting of the zymogen. The mechanisms of conversion to active enzymes are diverse in nature, ranging from enzymatic or nonenzymatic cofactors that trigger activation, to a simple change in pH that results in conversion by an autocatalytic mechanism. Recent X-ray crystallographic studies of zymogens and comparisons with their active counterparts have identified the structural changes that accompany conversion. This review will focus upon the structural basis for inhibition by activation segments, as well as the molecular events that lead to the conversion of zymogens to active enzymes.


Dihydroxy-, hydroxyspirolactone-, and dihydroxyspirolactone-urochlorins induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in the liver of mice.
Previous work has shown that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes porphyria, enhanced by iron, in C57BL/6J mice with marked accumulation in the liver of uroporphyrin I and III isomers and heptacarboxylic acid III and is one model of human porphyria cutanea tarda. Preliminary examination by HPLC also indicated the presence of some oxygenated side chain uroporphyrin derivatives. Here, the porphyrin constituents of TCDD-induced porphyric liver have been examined by HPLC/electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC/ESI-Q-TOFMS) to characterize the major and minor porphyrins present in hepatic tissue. As well as the major constituents uroporphyrins I and III, we identified the isomers of heptacarboxylic, hexacarboxylic, and pentacarboxylic acid porphyrins arising from intermediates in the stepwise decarboxylation of uroporphyrinogen I and III to coproporphyrinogens. In addition, monohydroxy analogues of uroporphyrin isomers were detected hydroxylated in the acetic acid and beta-positions of propionic acid side chains and in the meso ring position. Of particular note, for the first time for human and experimental porphyrias, we found chlorins (dihydroxy-, hydroxyspirolactone- ,and dihydroxyspirolactone-urochlorins) consistent with those derived from an epoxyurochlorin structure, formed by oxidation of the double bond of a pyrrole ring of uroporphyrinogen I and III isomers. The findings demonstrate that oxygen insertion into the pyrrole rings of uroporphyrinogens occurs under pathological circumstances in vivo and support the evidence for an oxidative cellular environment present in TCDD-treated porphyric tissue.


Allelic imbalance in sporadic parathyroid carcinoma and evidence for its de novo origins.
Parathyroid cancer is a rare, clinically aggressive cause of primary hyperparathyroidism, and whether these malignancies generally evolve from pre-existing benign adenomas or arise de novo is unclear. Furthermore, while inactivation of the CDC73 (HRPT2) tumor suppressor gene, encoding parafibromin, is a major contributor, other genes essential to parathyroid carcinogenesis remain unknown. We sought to identify genomic regions potentially harboring such oncogenes or tumor suppressor genes, and to gain insight into the origins and molecular relationship of malignant versus benign parathyroid tumors. We performed genome-wide copy-number and loss of heterozygosity analysis using Affymetrix 50K SNP mapping arrays and/or comparative genomic hybridization on 16 primary parathyroid carcinomas, local recurrences or distant metastases, and matched normal controls, from 10 individuals. Recurrent regions of allelic loss were observed on chromosomes 1p, 3, and 13q suggesting that key parathyroid tumor suppressor genes are located in these chromosomal locations. Recurrent allelic gains were seen on chromosomes 1q and 16, suggesting the presence of parathyroid oncogenes on these chromosomes. Importantly, the most common alteration in benign parathyroid adenomas, loss of 11q, was not found as a recurrent change in the malignant parathyroid tissues. Molecular allelotyping using highly polymorphic microsatellite markers provided further confirmation that the prevalence of 11q loss is markedly and significantly lower in carcinomas as compared with adenomas. Our observations provide molecular support for the concept that sporadic parathyroid cancer usually arises de novo, rather than evolving from a pre-existing typical benign adenoma. Furthermore, these results help direct future investigation to ultimately determine which of the candidate genes in these chromosomal locations make significant contributions to the molecular pathogenesis of parathyroid cancer.


d-amino acid oxidase promotes cellular senescence via the production of reactive oxygen species.
d-amino acid oxidase (DAO) is a flavin adenine dinucleotide (FAD)-dependent oxidase metabolizing neutral and polar d-amino acids. Unlike l-amino acids, the amounts of d-amino acids in mammalian tissues are extremely low, and therefore, little has been investigated regarding the physiological role of DAO. We have recently identified DAO to be up-regulated in cellular senescence, a permanent cell cycle arrest induced by various stresses, such as persistent DNA damage and oxidative stress. Because DAO produces reactive oxygen species (ROS) as byproducts of substrate oxidation and the accumulation of ROS mediates the senescence induction, we explored the relationship between DAO and senescence. We found that inhibition of DAO impaired senescence induced by DNA damage, and ectopic expression of wild-type DAO, but not enzymatically inactive mutant, enhanced it in an ROS-dependent manner. Furthermore, addition of d-amino acids and riboflavin, a metabolic precursor of FAD, to the medium potentiated the senescence-promoting effect of DAO. These results indicate that DAO promotes senescence through the enzymatic ROS generation, and its activity is regulated by the availability of its substrate and coenzyme.


The natural history diverticular disease: is there a role for elective colectomy?
The natural history of colonic diverticular disease is unclear leading to a debate on the value of elective colectomy in preventing complications of the disease. To assess whether the complications of diverticular disease requiring emergency surgery are related to previous episodes of diverticulitis and whether elective colectomy might prevent such complications. A retrospective study was done on all patients admitted with complicated diverticular disease in two adjacent district general hospitals between 1995 and 2000. Information was collected on the details of management of the complications and past history of the investigations and treatment for diverticular disease in these patients. A total of 108 patients were admitted with complicated diverticular disease. Ninety eight (91%) patients were admitted as an emergency for perforated diverticular disease and rectal bleeding. Ten patients were urgent admissions for fistulae and diverticular phlegmons. Ninety eight patients underwent a Hartmann's operation, two had a subtotal colectomy and 4 patients had a sigmoid colectomy. Thirty four (31.4%) patients died in hospital post-operatively. Of the 108 patients, only 28 (26%) patients were known to have diverticular disease previously. Only three (2.7%) patients had had an episode of acute diverticulitis before they presented with further complications. Complications of diverticular disease occur de novo in the majority of patients who have no previous history of the disease. Further studies are needed to identify risk factors for complicated diverticular disease before adopting a policy of elective interval colectomy.


The role of transperineal template biopsies of the prostate in the diagnosis of prostate cancer: a review.
The incidence of prostate cancer has shown a significant increase, highlighting the importance of early diagnosis. Current practice considers histological diagnosis a necessity in the majority of the cases. The limitations of transrectal biopsies led to the development of the promising transperineal prostatic biopsies. The latter offers a safer approach by avoiding the rectum, utilizing brachytherapy template grid to detect anterior zone disease and provides accurate prostatic mapping by systematically sampling the whole gland. It also helps to direct biopsies based on images obtained from previous prostate scanning and identify those eligible for focal therapy to direct focal treatment accurately. The current literature provides enough reassurance that transperineal template biopsies are effective, efficient and superior to the traditional and inaccurate transrectal biopsies. The absence of consensus on the technical aspect of template biopsies is a drawback, yet it highlights the need to develop robust guidelines to standardize the procedure.


Localization of amyloid beta peptides to locus coeruleus and medial prefrontal cortex in corticotropin releasing factor overexpressing male and female mice.
A culmination of evidence from the literature points to the Locus Coeruleus (LC)-Norepinephrine system as an underappreciated and understudied area of research in the context of Alzheimer's Disease (AD). Stress is a risk factor for developing AD, and is supported by multiple clinical and preclinical studies demonstrating that amplification of the stress system disrupts cellular and molecular processes at the synapse, promoting the production and accumulation of the amyloid beta (Aβ42) peptide. Stress-induced activation of the LC is mediated by corticotropin releasing factor (CRF) and CRF receptors exhibit sex-biased stress signaling. Sex differences are evident in the neurochemical, morphological and molecular regulation of LC neurons by CRF, providing a compelling basis for the higher prevalence of stress-related disorders such as AD in females. In the present study, we examined the cellular substrates for interactions between Aβ and tyrosine hydroxylase a marker of noradrenergic somatodendritic processes in the LC, and Dopamine-β-Hydroxylase (DβH) in the infralimbic medial prefrontal cortex (ILmPFC) in mice conditionally overexpressing CRF in the forebrain (CRFOE) under a Doxycycline (DOX) regulated tetO promoter. CRFOE was sufficient to elicit a redistribution of Aβ peptides in the somatodendritic processes of the LC of male and female transgenic mice, without altering total Aβ42 protein expression levels. DOX treated groups exhibited lysosomal compartments with apparent lipofuscin and abnormal morphology, indicating potential dysfunction of these Aβ42-clearing compartments. In female DOX treated groups, swollen microvessels with lipid-laden vacuoles were also observed, a sign of blood-brain-barrier dysfunction. Finally, sex differences were observed in the prefrontal cortex, as females responded to DOX treatment with increased frequency of co-localization of Aβ42 and DβH in noradrenergic axon terminals compared to vehicle treated controls, while male groups showed no significant changes. We hypothesize that the observed sex differences in Aβ42 distribution in this model of CRF hypersignaling is based on increased responsivity of female rodent CRFR1 in the LC. Aβ42 production is enhanced during increased neuronal activation, therefore, the excitation of DOX treated female CRFOE LC neurons projecting to the mPFC may exhibit more frequent co-localization with Aβ due to increased neuronal activity of noradrenergic neurons.


Association Between Insufficient Sleep, Depressive Symptoms, and Suicidality Among Florida High School Students.
Emerging research suggests insufficient sleep can heighten the risk of poor mental health among adolescents. We examined the relationship between sleep, depressive symptoms, and suicidal thoughts and behaviors among Florida high school students. Data were from the 2021 Florida High School Youth Risk Behavior Survey, a statewide representative sample of students in grades 9 to 12. We estimated prevalence of 1 indicator of depressive symptoms - feeling sad or hopeless for 2 or more weeks - and 2 indicators of suicidality - having considered attempting suicide and making a suicide plan - by insufficient sleep status (<8 h of sleep on an average school night). Multivariate logistic regressions were used to calculate odds ratios measuring the association between insufficient sleep, poor mental health, and suicidality. Compared with students reporting sufficient sleep, those with insufficient sleep were more likely to feel sad or hopeless (42.7% vs 28.1%), have considered suicide (19.1% vs 12.5%), or have made a suicide plan (14.8% vs 9.6%). In adjusted multivariate models, compared with students with sufficient sleep, students with insufficient sleep had higher odds of feeling sad or hopeless (adjusted odds ratio [AOR] = 1.83; 95% CI 1.54-2.17), and having made a plan about how they would attempt suicide (AOR = 1.32; 95% CI, 1.00-1.74), but did not have significantly higher odds of having seriously considered suicide. Increased attention to sleep as a modifiable risk factor for mental health among adolescents is particularly important because of the many less modifiable factors that contribute to mental health difficulties in this population. Adolescent mental health programs should consider sleep an important factor in suicide prevention.


[Comparison of the prevalence of cleft lip and/or palate in children in Lodz province in the period of 1996-2010 in 5-years intervals].
Clefts of lip and/or palate are the most common facial malformations. Taking into consideration complexity of this defect and necessity of multidisciplinary treatment the knowledge of actual scale of this phenomenon seems to be relevant. The aim of this paper it to describe and to compare the prevalence of cleft lip and/or palate in Lodz province in the period of 1996-2010 in 5 years intervals. MATERIAL AND METHODS; From Polish Register of Congenital Malformations we managed to obtain the data of the number of live born infants and birth of children with clefts in Lodz province in 2001-2010. On the basis of previously performed studies Department of Plastic, Reconstructive and Esthetic Surgery has the data of clefts' prevalence in Lodz province from year 1996 to 2000. The mean frequency of clefts was 1,67/1000 live born infants in 1996-2000, during years 2001-2005 - 1,47/1000, and 1,13/1000 in 2006-2010. Significant decrease of the prevalence of cleft lip and/or palate is observed in Lodz province in the period of 1996-2010. In boys the most frequent defect is cleft lip and palate, while in girls--isolated cleft palate.


Chemical composition, antimicrobial, and antioxidant cytotoxic activities of essential oil from Actinidia arguta.
Chemical composition, antimicrobial, antioxidant, and cytotoxic properties of Actinidia arguta essential oil (AEO) were evaluated. Gas chromatography-mass spectrometry analysis identified 56 chemical compounds, with the most abundant being Squalene (23.08%), γ-sitrostorol (8.10%), and β-Tocopherol (7.01%). Whereas the AEO had significant antimicrobial activity against Staphylococcus aureus and Saccharomyces cerevisiae, it showed mild efficacy against Bacillus subtilis and Microsporum canis. On the contrary, the Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, were not susceptible to the AEO pressure. On the other hand, the AEO exhibited strong antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), β-carotene, and hydroxyl radicals, with IC50 values of 117.60, 73.60, and 35.15 μg/mL, respectively. Additionally, compared to the PC-3 or HT-29 cell lines, the A549 cells were more susceptible to the AEO (IC50; 6.067 mg/mL). Besides, the confocal laser scanning microscopy imaging showed that 16 mg/mL of the AEO-induced apoptosis in the A549 cell lines. Our data indicated that the AEO might be useful in the food and pharmaceutical industry. Preparation of Actinidia arguta essential oil (a) and schematic overview of the experiment (b).


Highly fluorescent hyperbranched BODIPY-based conjugated polymer dots for cellular imaging.
The first hyperbranched BODIPY-based conjugated polymer dots (Pdots) were reported. The Pdots showed quantum yields of as high as 22%, which is 40% higher than their linear counterparts. The Pdots were successfully applied in cell-labelling applications. These results demonstrated that hyperbranched polymers are a very promising material for biological imaging and applications.


Distribution of a highly lipophilic drug cannabidiol into different lymph nodes following oral administration in lipidic vehicle.
Efficient delivery of highly lipophilic drugs or prodrugs to the mesenteric lymph nodes (MLN) can be achieved following oral administration with lipids. However, it remains unclear which specific MLN can be targeted and to what extent. Moreover, the efficiency of drug delivery to the retroperitoneal lymph nodes (RPLN) has not been assessed. The aim of this study was to assess the distribution of a highly lipophilic model drug cannabidiol (CBD), known to undergo intestinal lymphatic transport following administration with lipids, into specific MLN and RPLN in rats at various time-points post dosing. In vivo studies showed that at 2 h following administration, significantly higher concentrations of CBD were present in the region second from the apex of the MLN chain. From 3 h following administration, concentrations in all MLN were similar. CBD was also found at substantial levels in RPLN. This study demonstrates that drug concentrations in specific MLN are different, at least at the peak of the absorption process. Moreover, in addition to the MLN, the RPLN may also be targeted by oral route of administration, which may have further implications for treatment of a range of diseases.


Nicotine does not affect stem cell properties requisite for suicide gene therapy against glioma.
Glioblastoma is one of the most lethal tumors in adult central nervous system with a median survival of a year and half and effective therapeutic strategy is urgently needed. For that reason, stem cell-based suicide gene therapies have attracted much interest because of potent tumor tropism of stem cells and bystander effect. In this current clinical situation, stem cells are promising delivery tool of suicide genes for glioma therapy. Since habitual cigarette smoking still prevails worldwide, we investigated the effect of nicotine on stem cell tropism toward glioma and gap junctional intercellular communication (GJIC) function between glioma and stem cells, both of which are important for suicide gene therapies. Methods: Mouse induced pluripotent stem cell-derived neural stem cells (iPS-NSCs) and human dental pulp mesenchymal stem cells (DPSCs) were used. The effect of nicotine on tumor tropism to glioma-conditioned medium (CM) at a non-cytotoxic concentration was assessed with Matrigel invasion assay. Nicotine effect on GJIC was assessed with the scrape loading/dye transfer (SL/DT) assay for co-culture of glioma and stem cells and the parachute assay among glioma cells using high-content analysis. Results: Tumor tropism of iPS-NSCs toward GL261-CM and DPSCs toward U251-CM was not affected by nicotine (0.1 and 1 µM). Nicotine at the concentrations equivalent to habitual smoking (1 µM) did not affect GJIC of iPS-NSC/GL261 and DPSC/U251 and GJIC among each glioma cells. Conclusions: The study demonstrated that non-cytotoxic concentrations of nicotine did not significantly change the stem cell properties requisite for stem cell-based suicide gene therapy.