In vitro oxidized and glycated human low-density lipoprotein particles characterized by capillary zone electrophoresis.
A simple capillary zone electrophoresis (CZE) method was used to determine native, in vitro Cu(2+) and glucose modified low-density lipoprotein (LDL) particles for four healthy subjects. The LDL electropherograms are highly reproducible with good precisions of effective mobility and peak area. The native LDL capillary electrophoresis (CE) profile shows a major peak with lower mobility and two minor peaks with higher mobilities. For three-hour Cu(2+) oxidation, one major peak with mobility close to that of the native major peak, and one minor peak with mobility extending to -47 x 10(-5)cm(2)V(-1)s(-1) appear. For eighteen-hour Cu(2+) oxidation, one major peak with mobility much higher than that of the native major peak appears. As the reaction time for LDL and Cu(2+) increases from 0 to 24h, effective mobility of the LDL major peak increases, suggesting that LDL particles become more negatively charged and oxidized as the time increases. The in vitro glycated LDL particles are characterized by a major peak and two minor peaks. Mobility of the major peak is close to that of native major peak, but the second minor peak is much more negatively charged with mobility extending to -53 x 10(-5)cm(2)V(-1)s(-1). Native, oxidized and glycated LDL particles show distinctive differences in their CZE profiles. Agarose electrophoresis shows that the charge to mass ratios of native, three-hour Cu(2+) and glucose modified LDL particles are similar, but that of eighteen-hour Cu(2+) oxidized LDL particles is higher.


Metabolism of exogenously administered natural surfactant in the newborn lamb.
[3H]-Palmitate labeled natural lamb surfactant and free [14C]-choline were mixed with the lung fluid of 11 term lambs at cesarean section, before the first breath. After receiving the isotope, the lambs were delivered, allowed to breathe spontaneously, and were subsequently sacrificed from 5 min to 96 h of age. Alveolar washes, lung homogenates, microsomal and lamellar body fractions of lungs, and pulmonary alveolar macrophages were examined for the presence of labeled phosphatidylcholine. Analysis of the labeled natural surfactant kinetic data revealed an apparent t 1/2 of phosphatidylcholine in the whole lung of 6.0 days. This half-life can be interpreted only as a rough estimate. Appearance of considerable [3H] labeled phosphatidylcholine in the lung homogenates demonstrated uptake of phosphatidylcholine from alveoli into lung tissue. The surfactant-associated label in homogenates was localized preferentially to lamellar body fractions. Some of the administered [14C]-choline appeared in phosphatidylcholine. Almost all of this labeled phosphatidylcholine was associated with the homogenate. Extremely small % of administered [3H] and [14C] were found in pulmonary alveolar macrophages.


Design, synthesis and biological evaluation of substituted flavones and aurones as potential anti-influenza agents.
We designed a series of substituted flavones and aurones as non-competitive H1N1 neuraminidase (NA) inhibitors and anti-influenza agents. The molecular docking studies showed that the designed flavones and aurones occupied 150-cavity and 430-cavity of H1N1-NA. We then synthesized these compounds and evaluated these for cytotoxicity, reduction in H1N1 virus yield, H1N1-NA inhibition and kinetics of inhibition. The virus yield reduction assay and H1N1-NA inhibition assay demonstrated that the compound 1f (4-methoxyflavone) had the lowest EC50 of 9.36 nM and IC50 of 8.74 μM respectively. Moreover, kinetic studies illustrated that compounds 1f and 2f had non-competitive inhibition mechanism.


The oncological multidimensional prognostic index is a promising decision-making tool: A real-world analysis in older patients with metastatic colorectal cancer.
Approximately 50% of colorectal cancers occur in older patients. International societies recommend geriatric tools to optimise treatment of older patients. Comprehensive Geriatric Assessment (CGA) is a multidimensional assessment used to classify patients as fit, vulnerable, or frail. The CGA-based oncological multidimensional prognostic index (onco-MPI) also classifies patients as high-, intermediate-, or low-risk based on tumour characteristics. We investigated the role of CGA and onco-MPI in older patients with metastatic colorectal cancer (mCRC) in a real-world setting. Data for consecutive mCRC patients aged ≥70 years were retrieved from a prospectively maintained database from 2010 to 2020. We analyzed patients' and tumours' characteristics, and the CGA domains. Onco-MPI was calculated by a validated algorithm derived from CGA domains. Pearson's test was used to verify whether onco-MPI scores and chemotherapy administration were correlated. The study included 488 mCRC patients with a mean age of 76.1 years. According to CGA, 52% of patients were fit, 28% vulnerable, and 20% frail. According to onco-MPI, 9% were low, 54% intermediate, and 37% high-risk. The median OS was 22.7 months. The following factors improved OS: 0-1 ECOG PS, low onco-MPI, fit based on CGA, chemotherapy administration, and doublet regimen. Chemotherapy administration significantly correlated with onco-MPI scores, leading to a survival gain regardless of the risk subgroups. First-line regimen had no impact on survival across the CGA and onco-MPI categories. CGA and onco-MPI scores confirmed their prognostic impact in older mCRC patients and may aid in decision-making and subgroup stratification in dedicated trials.


Ultrasound prediction of fetal weight in prolonged pregnancy.
In this study we applied two commonly used birth weight prediction equations to a sample of 121 women with prolonged pregnancies. Subjects had sonographic measurements of biparietal diameter and abdominal perimeter taken within 2 days of delivery at Parkland Memorial Hospital. Although the two prediction equations were obtained from a population of women in New Haven, Connecticut, who delivered over a wide range of gestational ages, when the equations were applied to the sample of prolonged pregnancies in Dallas, Texas, there was a strong correlation (0.71) between predicted and actual birth weight. Moreover, reestimation of the New Haven equations with use of the Dallas data yielded similar regression coefficients. Finally, birth weight prediction equations for black, white, and Hispanic patients in Dallas were not significantly different. These findings suggest a remarkably constant relationship between fetal head and abdominal dimensions and birth weight over different gestational ages and for different population groups.


3-Chloro-1,2-propanediol biodegradation by Ca-alginate immobilized Pseudomonas putida DSM 437 cells applying different processes: mass transfer effects.
3-Chloro-1,2-propanediol (3-CPD) biodegradation by Ca-alginate immobilized Pseudomonas putida cells was performed in batch system, continuous stirred tank reactor (CSTR), and packed-bed reactor (PBR). Batch system exhibited higher biodegradation rates and 3-CPD uptakes compared to CSTR and PBR. The two continuous systems (CSTR and PBR) when compared at 200 mg/L 3-CPD in the inlet exhibited the same removal of 3-CPD at steady state. External mass-transfer limitations are found negligible at all systems examined, since the observable modulus for external mass transfer Ω ≪ 1 and the Biot number Bi > 1. Intra-particle diffusion resistance had a significant effect on 3-CPD biodegradation in all systems studied, but to a different extent. Thiele modulus was in the range of 2.5 in batch system, but it was increased at 11 when increasing cell loading in the beads, thus lowering significantly the respective effectiveness factor. Comparing the systems at the same cell loading in the beads PBR was less affected by internal diffusional limitations compared to CSTR and batch system, and, as a result, exhibited the highest overall effectiveness factor.


Photolabeling of calmodulin with basic, amphiphilic alpha-helical peptides containing p-benzoylphenylalanine.
A novel photoreactive amino acid has been incorporated synthetically into two model peptides and the calmodulin-binding domain from myosin light chain kinase. Cross-linked photoadducts of each peptide with calmodulin have been prepared and digested by chemical and/or enzymatic methods to determine the site of label attachment. Depending on the position of the photoprobe in the peptide sequence, either Met-144 or Met-71 is photolabeled. These results are discussed in relation to the three-dimensional structure of calmodulin obtained crystallographically and the known solution properties of calmodulin.


Morphological and molecular characterizations of Sarcocystis miescheriana and Sarcocystis suihominis in domestic pigs (Sus scrofa) in China.
In this study, 36.8% (28/76) of tissue samples collected from domestic pigs (Sus scrofa) contained sarcocysts, as determined by light microscopy. The organisms were identified as Sarcocystis miescheriana and Sarcocystis suihominis based on their morphological and molecular characteristics. Four genetic markers, i.e., 18S rDNA, 28S rDNA, ITS-1 region (ITS-1), and the mitochondrial COX1 gene (COX1), of the two parasites were sequenced and analyzed, and the 28S rDNA and ITS-1 of S. suihominis obtained from pigs constituted the first records of these markers in GenBank. The sequences of the four loci (18S rDNA, 28S rDNA, ITS-1, and COX1) of S. miescheriana shared high identities with those of S. miescheriana obtained from domestic and/or wild pigs in GenBank, with similarities of 99.6%, 99.6%, 95.9%, and 95.4%, respectively. The 18S rDNA sequences of S. suihominis exhibited 99.4% identity with those of S. suihominis from domestic and wild pigs. The comparison of the newly obtained sequences of the four genetic markers between the two parasites revealed that the interspecific similarities of 18S rDNA, 28S rDNA, ITS-1, and COX1 were 97.7%, 96.6%, 80.3%, and 81.2%, respectively. Therefore, the two species could be better discriminated with ITS-1 and mitochondrial COX1 compared with 18S rDNA or 28S rDNA. The phylogenetic analysis using 28S rDNA indicated that the two Sarcocystis species in domestic pigs had a close relationship.


Extracellular vesicles in respiratory disease.
Extracellular vesicles (EVs) are membranous nanoparticles secreted by nearly all cell types and play a critical role in cell-to-cell crosstalk. EVs can be categorized based on their size, surface markers, or the cell type from which they originate. EVs carry "cargo," including but not limited to, RNA, DNA, proteins, and small signaling molecules. To date, many methods have been developed to isolate EVs from biological fluids, such as blood plasma, urine, bronchoalveolar lavage fluid, and urine. Once isolated, EVs can be characterized by dynamic light scattering, nanotracking analysis, nanoscale flow cytometry, and transmission electron microscopy. Given the ability of EVs to transport cargo between cells, research has recently focused on understanding their role in various human diseases. As understanding of their significance to disease processes grows, insight into the mechanisms behind the physiological role of their cargo in target cells can facilitate the development of a new type of biomarker and therapeutic target for diseases in future. In addition, their ability to deliver their cargo selectively to target cells within the human body means that they could serve as therapeutic agents or methods of drug delivery. In this review, we will first introduce EVs and the cargo they carry, outline current methods for EV isolation and characterization, and discuss their potential use as biomarkers and therapeutic agents in the near future.


Strict selection criteria in uncontrolled donation after circulatory death provide excellent long-term kidney graft survival.
We aimed to report our experience in uncontrolled donation after circulatory death (uDCD) kidney transplantation applying a strict donor selection and preservation criteria. All kidney recipients received a graft from a local uDCD. As controls, we included all renal transplants from local standard criteria donation after brain death (SDBD) donors. Normothermic regional perfusion was the preservation method in all cases. A total of 19 kidneys from uDCD donors were included and 67 controls. Delayed graft function (DGF) was higher in the uDCD group (42.1% vs 17.9%; P = .033), whereas no differences were observed in primary nonfunction (0% cases vs 3% controls; P = .605). The estimated glomerular filtration rate was identical in both groups. No differences were observed in graft survival censored for death between the uDCD and the SDBD groups at 1-year (100% vs 95%) or 5-year follow-up (92% vs 91%). uDCD kidney recipients did not have higher risk of graft loss in the multivariate analysis adjusted by recipient age, cold ischemic time, presence of DGF, and second kidney transplant (HR: 0.4; 95% CI 0.02-6; P = .509). Obtaining renal grafts from uDCD is feasible in a small city and provides similar outcomes compared to standard DBD donors.


Silent coronary artery-left ventricular fistula: a disorder of the thebesian system?
Two cases with abnormal electrocardiogram were found to have the unusual direct communication between the coronary artery and left ventricular chamber without any manifestations of the other reported coronary arterial fistula.


EPR spectra of photosystem I and other iron protein components in intact cells of cyanobacteria.
Electron paramagnetic resonance (EPR) spectra were recorded of whole filaments of the cyanobacteria Nostoc muscorum and Anabaena cylindrica. Signals due to manganese were removed by freezing and thawing the cells in EDTA. EPR spectra were assigned on the basis of their g values, linewidths, temperature dependence and response to dithionite and light treatments. The principal components identified were: (i) rhombic Fe3+ (signal at g = 4.3), probably a soluble storage form of iron; (ii) iron-sulfur centers A and B of Photosystem I; (iii) the photochemical electron acceptor 'X' of Photosystem I; this component was also observed for the first time in isolated heterocysts; (iv) soluble ferredoxin which was present at a concentration of 1 molecule per 140 +/- 20 chlorophyll molecules; (v) a membrane-bound iron-sulfur protein (g = 1.92). A signal g = 6 in the oxidized state was probably due to an unidentified heme compound. During deprivation of iron the rhombic Fe3+, centers A, B and X of Photosystem I, and soluble ferredoxin were all observed to decrease.


Apolipoprotein CIII hyperactivates β cell CaV1 channels through SR-BI/β1 integrin-dependent coactivation of PKA and Src.
Apolipoprotein CIII (ApoCIII) not only serves as an inhibitor of triglyceride hydrolysis but also participates in diabetes-related pathological events such as hyperactivation of voltage-gated Ca(2+) (CaV) channels in the pancreatic β cell. However, nothing is known about the molecular mechanisms whereby ApoCIII hyperactivates β cell CaV channels. We now demonstrate that ApoCIII increased CaV1 channel open probability and density. ApoCIII enhanced whole-cell Ca(2+) currents and the CaV1 channel blocker nimodipine completely abrogated this enhancement. The effect of ApoCIII was not influenced by individual inhibition of PKA, PKC, or Src. However, combined inhibition of PKA, PKC, and Src counteracted the effect of ApoCIII, similar results obtained by coinhibition of PKA and Src. Moreover, knockdown of β1 integrin or scavenger receptor class B type I (SR-BI) prevented ApoCIII from hyperactivating β cell CaV channels. These data reveal that ApoCIII hyperactivates β cell CaV1 channels through SR-BI/β1 integrin-dependent coactivation of PKA and Src.


Coarse-graining time series data: Recurrence plot of recurrence plots and its application for music.
We propose a nonlinear time series method for characterizing two layers of regularity simultaneously. The key of the method is using the recurrence plots hierarchically, which allows us to preserve the underlying regularities behind the original time series. We demonstrate the proposed method with musical data. The proposed method enables us to visualize both the local and the global musical regularities or two different features at the same time. Furthermore, the determinism scores imply that the proposed method may be useful for analyzing emotional response to the music.


Fat elimination in acute renal failure.
Lipid metabolism and elimination of parenterally administered fat were investigated in 15 patients with acute renal failure (ARF). The mean triglyceride level was elevated to 2.56 +/- 1.43 mmol/l and the mean cholesterol level was 3.32 +/- 0.66 mmol/l, which is slightly below the normal range. A type IV hyperlipoproteinaemia was present in 47 per cent of the patients. The triglyceride content of LDL and VLDL was elevated and the cholesterol concentration of HDL and of LDL was reduced markedly. The fractional removal rate of triglycerides (K2) evaluated by an intravenous fat tolerance test using a bolus technique was reduced to 2.44 +/- 1.56 per cent/min which is about half of normal and correspondingly the elimination half life was prolonged to 28.4 min. No correlation could be demonstrated between the impairment of fat elimination and residual renal function, basal and VLDL triglyceride concentration or HDL cholesterol content.


Risk factors for perioperative mortality and transfusion in sacrococcygeal teratoma resections.
Sacrococcygeal teratomas are a common congenital tumor. Surgical resection can occur in utero, in the neonatal period, or in the postneonatal period. We describe patient and tumor factors associated with mortality and transfusion in this population. We did a retrospective chart review of patients who underwent sacrococcygeal teratoma resection between January 1998 and March 2016. Demographic data, transfusion data, and tumor characteristics were collected. Descriptive statistics were calculated, and univariate comparisons were performed with chi-square test and Fisher's exact test. Variables significant at univariate level were used in multivariate logistic regression and negative binomial regression. Of the 112 cases, 6 were in utero repairs, 73 were neonatal repairs, and 33 were repairs at >30 days of life. There was 17%, 1%, and 0% intraoperative mortality and 33%, 5%, and 0% 30-day mortality in the in utero, neonatal, and >30 days of life repairs, respectively. All six patients who died within the first 30 days of life had a postmenstrual age of <32 weeks at time of surgery. All six patients who died had noncystic tumors. Patients with noncystic tumors were more likely to be born prior to 30-week gestation (23/65 vs 6/47; χ2 = 7.3; P = 0.007). Gestational age >30 weeks was associated with decreased intraoperative death (0% vs 10%; modified maximum likelihood estimate of OR 0.05; 95% CI 0.002-0.96; P = 0.02). Gestational age >30 weeks (2.4% vs 13.8%; OR 0.15; 95% CI 0.03-0.89; P = 0.04) and cystic morphology (0% vs 9.2%; modified maximum likelihood estimate of OR 0.1; CI 0.01-1.75; P = 0.04) were associated with decreased 30-day mortality and emergent surgery (17.9% vs 1.2%; OR 18; 95% CI 2-162.2; P = 0.004) was associated with increased 30-day mortality. Gestational age >30 weeks (33.7% vs 62.1%; OR 0.27; 95% CI 0.09-0.79; P = 0.02) and Altman class 3-4 (12.1% vs 52.7%; OR 0.1; 95% CI 0.03-0.34; P = 0.0002) were associated with decreased need for transfusion and noncystic tumor was associated with increased transfusion volume (131.6 ml·kg-1 [95% CI 94-184] vs 63 ml·kg-1 [95% CI 40-100.1]; P = 0.01). Prematurity is associated with increased intraoperative and 30-day mortality. Noncystic tumor morphology was the only significant factor associated with transfusion volume and all six patients who died had transfusion volumes of 240 ml·kg-1 or greater. In these patients at high risk of mortality due to blood loss, the anesthesia team should be prepared to manage massive transfusion and coagulopathy with blood components and pharmacologic measures.


Structural independence of hydrogen-bond symmetrisation dynamics at extreme pressure conditions.
The experimental study of hydrogen-bonds and their symmetrization under extreme conditions is predominantly driven by diffraction methods, despite challenges of localising or probing the hydrogen subsystems directly. Until recently, H-bond symmetrization has been addressed in terms of either nuclear quantum effects, spin crossovers or direct structural transitions; often leading to contradictory interpretations when combined. Here, we present high-resolution in-situ 1H-NMR experiments in diamond anvil cells investigating a range of systems containing linear O-H ⋯ O units at pressure ranges of up to 90 GPa covering their respective H-bond symmetrization. We found pronounced minima in the pressure dependence of the NMR resonance line-widths associated with a maximum in hydrogen mobility, precursor to a localisation of hydrogen atoms. These minima, independent of the chemical environment of the O-H ⋯ O unit, can be found in a narrow range of oxygen oxygen distances between 2.44 and 2.45 Å, leading to an average critical oxygen-oxygen distance of [Formula: see text] Å.


Fixed versus Flexible Gonadotropin Releasing Hormone Antagonist Protocol in Controlled Ovarian Stimulation for Invitro Fertilization in Women with Polycystic Ovary Syndrome.
This study was conducted to compare the results of fixed versus flexible GnRH antagonist protocols in controlled ovarian stimulation for Intra Cytoplasmic Sperm Injection (ICSI) in patients with PCOS. A randomized clinical trial was performed on 100 PCOS women, who were admitted to a tertiary infertility clinic and were candidate for IVF/ICSI. They were divided into two groups based on the GnRH antagonist protocol. We started GnRH antagonist 0.25mg in flexible protocol when a follicle ≥ 14 mm in diameter was seen in transvaginalsonography (Group 1). In fixed protocol, GnRH antagonist was administered from day 6 of stimulation (Group 2). Number of oocytes in methapase 2, number of developed and frozen embryo as main outcome and days of stimulation, number of gonadotropin and antagonist used assecondry outcome measures were assessed and compared between the two groups. The days of stimulation and the number of antagonist used was not significantly different between fixed and flexible group (p ≥ 0.05).Although the number of gonadotropin injections was significantly lower in flexible group (p = 0.03), the number of oocyte retrieved and the number of embryo which cryopreserved was significantly higher in flexible compared to fixed protocol (p < 0.01). It seems using flexible antagonist protocol in PCOS infertile patients is in favor of better outcomes in terms of number of good quality oocytes and embryo and possibility for cryopreservation for future cycles.


Descriptions of New Species of Chespirito Ferreira, Keller amp; Branham (Coleoptera: Lampyridae: Chespiritoinae) and the First Record for the Subfamily in the United States.
Three new species of Chespirito Ferreira, Keller and Branham 2020 are described from North America, including the first species from the United States: Chespirito milleri new species from Arizona, USA, Chespirito hintoni new species from Mexico state, Mexico and Chespirito costae new species from Morelo state, Mexico. An updated distribution map and key to males of Chespirito is provided along with illustrations of diagnostic characters of the newly described species.


[Diversity of signal transduction pathways which induce apoptosis of thymocytes. Role of bcl-2, pim-1, c-myc and p53 genes in selection processes of thymocytes].
There are independent signaling pathways which transmit apoptotic signals in thymocytes. c-Myc and p53 proteins participate in the apoptosis induction, whereas Bcl-2 and Pim-1 proteins inhibit complex apoptotic machinery.


Mode of action of butylated hydroxyanisole (BHA) and other phenols in preventing loss of 11 beta-hydroxylase activity in cultured bovine adrenocortical cells.
When cultured bovine adrenocortical cells are incubated with cortisol, or other steroids that are pseudosubstrates for 11 beta-hydroxylase (cytochrome P-45011 beta), the activity of the enzyme decreases. In previous experiments, three substances were shown to protect 11 beta-hydroxylase against loss of enzymatic activity in the presence of pseudosubstrates:BHA (butylated hydroxyanisole,2(3)-tert-butyl-4-methoxyphenol), dimethyl sulfoxide (DMSO), and metyrapone. The present experiments examine the protective effects of several phenolic analogs of BHA in this system, and compare their activities to that of DMSO and metyrapone. When a variety of analogs of BHA were tested for their abilities to prevent loss of 11 beta-hydroxylase activity in cultured adrenocortical cells incubated with 50 microM cortisol for 24 hr, phenol itself was found to be about equipotent with BHA. Addition of methyl, methoxy and benzyl groups to phenol did not diminish protective activity of the compound, but addition of one and particularly two tert-butyl groups greatly diminished activity. Thus, BHT(2,6-di-t-butyl-4-methylphenol) was inactive, in contrast to BHA. The hydroxy group of phenol was essential since benzene and fluorobenzene were inactive. Compounds with multiple hydroxyl groups were not as active as phenol itself, with the exception of catechol. No products of phenol formed during incubations of cells with cortisol were detected by high performance liquid chromatography. Estimated EC50 values for protection of 11 beta-hydroxylase by phenols were about 100 microM, whereas the EC50 values for dimethyl sulfoxide and metyrapone were 10 mM and 300 nM respectively. On a semilogarithmic plot, the dose-response curves for all these compounds were approximately parallel. To aid in determining the mechanism of protection of 11 beta-hydroxylase, phenols and DMSO were tested for prevention of loss of 11 beta-hydroxylase activity at three different oxygen concentrations (2, 5, and 19% O2). Lowering the oxygen concentration itself resulted in a small diminution of the loss of 11 beta-hydroxylase. Phenols and dimethyl sulfoxide were more effective at low oxygen and less effective in air. Because the cytochrome P-450 inhibitor metyrapone was found previously to be very effective in protecting 11 beta-hydroxylase against loss of activity, we examined whether phenols and dimethyl sulfoxide may act by directly inhibiting 11 beta-hydroxylase activity. In a 1-hr incubation with cells, BHA, phenol, and dimethyl sulfoxide all inhibited 11 beta-hydroxylase, but at concentrations that ranged from 4- to greater than 100-fold higher than those required for protection.(ABSTRACT TRUNCATED AT 400 WORDS)


Dexamethasone added to levobupivacaine prolongs the duration of interscalene brachial plexus block and decreases rebound pain after arthroscopic rotator cuff repair.
It has been reported that the addition of dexamethasone to interscalene brachial plexus block (ISBPB) prolongs the duration of the block effect. However, there have been no studies focusing on the effects of dexamethasone on rebound pain after the block effect has worn off. The aim of this study was to investigate the effect on postoperative pain when dexamethasone was added to ISBPB for arthroscopic rotator cuff repair (ARCR). In this multicenter, single-blinded, and randomized controlled study, 54 patients (33 males, 21 females) who received ARCR were randomly assigned to group L (ISBPB with 20 cc of 0.25% levobupivacaine; 21 patients) or group LD (ISBPB with 20 cc of 0.25% levobupivacaine + 3.3 mg dexamethasone; 33 patients). The primary outcome was the visual analog scale (VAS) for pain after the block effect had worn off. Secondary outcomes were the duration of analgesia, the time to the first request for additional analgesic, the number of additional doses of analgesic, and complications. The VAS scores on postoperative days 0 and 1 were significantly lower in group LD than group L (P = .005, .035). This indicated that the rebound pain was relieved in group LD. After postoperative day 1, there was no significant difference in VAS score (P = .43 and .19 for days 2 and 3, respectively). The duration of analgesia was significantly longer in group LD than group L (P < .001). The time to the first request for additional analgesic was significantly longer in group LD than group L (P < .001). The number of additional doses of analgesic was significantly lower in group LD (P < .001). In ARCR, the addition of dexamethasone to levobupivacaine not only prolongs the duration of ISBPB but also relieves rebound pain after the block effect wears off.


Dr Frederick Knight Hunt (1814-54) re-visited: medical man and journalist.
This paper studies Knight Hunt's career after he lost control of the Medical Times, and shows how he was brought to the notice of Charles Dickens who made him a sub-editor on the Daily News in 1845 and subsequently selected him as a pivotal contributor to Household Words, a post he gave up only when he became editor of the newspaper in 1851.


Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation.
Two genomic clones (lambda Ubi-1 and lambda Ubi-2) encoding the highly conserved 76 amino acid protein ubiquitin have been isolated from maize. Sequence analysis shows that both genes contain seven contiguous direct repeats of the protein coding region in a polyprotein conformation. The deduced amino acid sequence of all 14 repeats is identical and is the same as for other plant ubiquitins. The use of transcript-specific oligonucleotide probes shows that Ubi-1 and Ubi-2 are expressed constitutively at 25 degrees C but are inducible to higher levels at elevated temperatures in maize seedlings. Both genes contain an intron in the 5' untranslated region which is inefficiently processed following a brief, severe heat shock. The transcription start site of Ubi-1 has been determined and a transcriptional fusion of 0.9 kb of the 5' flanking region and the entire 5' untranslated sequence of Ubi-1 with the coding sequence of the gene encoding the reporter molecule chloramphenicol acetyl transferase (CAT) has been constructed (pUBI-CAT). CAT assays of extracts of protoplasts electroporated with this construct show that the ubiquitin gene fragment confers a high level of CAT expression in maize and other monocot protoplasts but not in protoplasts of the dicot tobacco. Expression from the Ubi-1 promoter of pUBI-CAT yields more than a 10-fold higher level of CAT activity in maize protoplasts than expression from the widely used cauliflower mosaic virus 35S promoter of a 35S-CAT construct. Conversely, in tobacco protoplasts CAT activity from transcription of pUBI-CAT is less than one tenth of the level from p35S-CAT.


Physiological responses of food neophobics and food neophilics to food and non-food stimuli.
Individual differences in human food neophobia (the reluctance to try novel foods) and food neophilia (the overt willingness to try novel foods) influence the evaluation of tastes and odors, as well as the sampling of such stimuli. Past research also notes an association of food neophobia to PTC sensitivity, body weight, and cephalic phase salivary response. The present study assessed physiological reactions of food neophobics and neophilics to pictures of food and non-food stimuli. Stimuli pictures were presented in random order on a computer screen for a period of 5 min. No significant differences were found between the groups in relation to non-food stimuli. However, pulse, GSR, and respirations were significantly increased in food neophobics when presented pictures of food stimuli. Thus, further evidence is provided to support a physiological component at least partially responsible for differences noted between neophobics and neophilics in sensitivity, psychophysical ratings, and "willingness to try" personality. Such a component may also lead to differences in weight, nutrition, and overall health.


Expression of platelet-derived growth factor (PDGF) and PDGF receptor genes in the developing rat brain.
Steady-state levels of rat central nervous system (CNS) platelet-derived growth factor (PDGF) A- and B-chain mRNAs were measured by a polymerase chain reaction method employing a synthetic gene internal standard, and the rates of transcription of PDGF A- and B-chain genes in CNS were estimated by a nuclear runoff assay. The abundance of PDGF B-chain mRNA was an order of magnitude below that of PDGF A-chain mRNA, while the rate of PDGF B-chain transcription was only slightly below that for the PDGF A-chain gene, indicating that the half-life of PDGF B-chain in CNS is shorter than that of PDGF A-chain mRNA. No developmental alterations in expression of the PDGF A- and B-chain genes were detected. By contrast, Northern blots showed that steady-state levels of mRNAs encoding the two PDGF receptor proteins, alpha and beta, were markedly higher in embryonic day 15 and postnatal day 6 rat brains than in later life. These results suggest that the actions of PDGF on the brain in vivo are regulated not at the level of PDGF A and B-chain gene expression, but rather by changes in the level of expression of PDGF alpha- and beta-receptor genes.


Microbiological transformation of two labdane diterpenes, the main constituents of Madia species, by two fungi.
Microbial transformation of 13R,14R,15-trihydroxylabd-7-ene (5) and 13R,14R,15-trihydroxylabd-8(17)-ene (6) by the fungus Debaryomyces hansenii gave 1 (13R,14R,15-trihydroxy-6-oxolabd-8-ene) and 3 (7alpha,13R,14R,15-tetrahydroxy-labd-8(17)-ene), respectively. While, microbial transformation of 5 by Aspergillus niger afforded 2 (3beta,13R,14R,15-tetrahydroxy-labd-7-ene), and 13R,14R,15-trihydroxylabd-8,17-ene (6) gave 3 and 4 (3R,14R,15-3-oxotetrahydroxy-labd-7-ene). The structures of the new compounds, 1 and 2, were assigned by 1D and 2D high-field NMR spectroscopic methods. Antimicrobial activity of these compounds were tested and their MIC were determined.


Potentiation of the effects of GABA by pentobarbitone.
Pentobarbitone (50 microM) has been shown to potentiate GABA-induced responses in rat isolated superior cervical ganglia and in dorsal or ventral root fibres of immature rat isolated spinal cords. Measurements of dose ratios for the antagonism of GABA-induced responses of dorsal root fibres by bicuculline showed that pentobarbitone (50 microM) did not significantly alter this antagonism. This result indicates that the antagonism of GABA by bucuculline at dorsal root fibres is not reversed by pentobarbitone. Ventral root responses to GABA (but not glycine) were also potentiated by pentobarbitone (+/-)-Nipecotic acid (300 microM) potentiated responses to GABA much more than those to 3-aminopropane-sulphonic acid.


Fast nucleotide identification through fingerprinting using gold nanoparticle-based surface-assisted laser desorption/ionisation.
We report a method centred on gold nanoparticle-based surface-assisted laser desorption/ionisation for analysis of deoxynucleotides and alkylated nucleobases. Gold nanoparticles allow for enhanced analysis capability by eliminating undesired signature peaks; thus more elegant mass spectra can be attained that allow identification by nucleotide mass fingerprint. The resulting fingerprinting patterns on the spectra are compared and associated with the presence of different nucleotides in the sample. This method can be easily extended to modified nucleotides implicated in genome lesions due to exposure to environment chemicals, such as DNA adducts (e.g. guanine adducts). The use of gold nanoparticles for surface-assisted laser desorption/ionisation can be an useful tool to resolve common issues of background noise when analysing nucleic acids samples.


Influence of azadirachtin, a botanical insecticide, on Chrysoperla carnea (Stephens) reproduction: toxicity and ultrastructural approach.
A commercial formulation of azadirachtin (Align) was tested on Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) adults to determine its effects on reproduction. Three methods of exposure were carried out in the laboratory (residual, direct contact and ingestion) with concentrations close to the maximum field recommended concentration in Spain (48 mg AI/liter). Align proved to be harmless to newly emerged adults irrespective of the mode of exposure. Nevertheless, fecundity was reduced in a reversible way in females that have ingested azadiracthin after the onset of oviposition. Fertility was not different from that of controls in all treatments. Additional tests showed that males were not involved in the reduction of oviposition. Electron microscopy studies pointed out that Align interfered with vitellogenin synthesis and/or its uptake by developing oocytes; thus, growing follicles in treated females were significantly smaller that those of the controls. The role of azadirachtin as a compound that may influence several hormonal and protein titers involved in reproduction is also discussed.


Role and mechanism of the AMPK pathway in waterborne Zn exposure influencing the hepatic energy metabolism of Synechogobius hasta.
Previous studies have investigated the physiological responses in the liver of Synechogobius hasta exposed to waterborne zinc (Zn). However, at present, very little is known about the underlying molecular mechanisms of these responses. In this study, RNA sequencing (RNA-seq) was performed to analyse the differences in the hepatic transcriptomes between control and Zn-exposed S. hasta. A total of 36,339 unigenes and 1,615 bp of unigene N50 were detected. These genes were further annotated to the Nonredundant protein (NR), Nonredundant nucleotide (Nt), Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups (COG) and Gene Ontology (GO) databases. After 60 days of Zn exposure, 708 and 237 genes were significantly up- and down-regulated, respectively. Many differentially expressed genes (DEGs) involved in energy metabolic pathways were identified, and their expression profiles suggested increased catabolic processes and reduced biosynthetic processes. These changes indicated that waterborne Zn exposure increased the energy production and requirement, which was related to the activation of the AMPK signalling pathway. Furthermore, using the primary hepatocytes of S. hasta, we identified the role of the AMPK signalling pathway in Zn-influenced energy metabolism.


[Familial periodic paralysis with hypokaliemia, hyperaldosteronism and extracellular vacuolization (author's transl)].
The observation of a 20 years old patient with a periodic paralysis is reported. During a crisis induced by 20 U of insulin and 150 g of glucose, transient hyperaldosteronism was found and the muscular biopsy performed in the left biceps brachii, disclosed vacuoles with a double membrane. The significance of these findings is discussed.


How subjective are subjective language complaints.
Subjective language complaints (SLC) are common during ageing but have not been investigated in detail. We aim to determine their association with demographic and clinical variables and objective cognitive performance. A sample of 479 individuals aged 50 years or above (average 66 ± 9.1 years), followed in primary care, with no history of brain disorder were asked two questions concerning SLC, fulfilled a depression scale and undertook a battery of cognitive tests. Response to questions regarding proper name retrieval (PNR) and word finding difficulties (WFD) was studied and their contribution to each measure of the battery calculated by repeated linear regression analysis, adjusting for age, sex, education, living alone and depressive symptoms. Word finding difficulties (47.6%) were more frequently reported than problematic PNR (10.9%). Both were more common in women, in subjects with depressive symptoms and in those living alone, but were unrelated with age or education. Both symptoms contributed significantly to the variance in tests of semantic fluency and episodic memory. PNR was also associated with immediate phonological memory. Subjective language complaints are especially common amongst individuals living alone and/or with depressive symptoms. They are associated with a worse cognitive performance in some memory and language-executive tests. Further studies are needed to understand their predictive value for cognitive decline.


Immediate adenoidectomy vs initial watchful waiting strategy in children with recurrent upper respiratory tract infections: an economic evaluation.
To compare the costs associated with 2 clinical strategies in children with recurrent upper respiratory tract infections (URTIs): immediate adenoidectomy vs an initial watchful waiting strategy. A cost-minimization analysis from a societal perspective including both direct and indirect costs, alongside an open randomized controlled trial with a 2-year follow-up. Multicenter study, including 11 general and 2 university hospitals in the Netherlands. The study population comprised 111 children aged 1 through 6 years, selected for adenoidectomy for recurrent URTIs according to current clinical practice. A strategy of immediate adenoidectomy with or without myringotomy or a strategy of initial watchful waiting. Difference in median costs during the 2-year follow-up. The median total of direct and indirect costs in the adenoidectomy and watchful waiting group were €1385 (US $1995) and €844 (US $1216) per patient, respectively. The extra costs in the adenoidectomy group are primarily attributable to surgery and visits to the otorhinolaryngologist. Other costs did not differ significantly between the groups. In children selected for adenoidectomy for recurrent URTIs, immediate adenoidectomy results in an increase in costs, whereas it confers no clinical benefit over an initial watchful waiting strategy. trialregister.nl Identifier:NTR968; isrctn.org Identifier:ISRCTN03720485.


The transcription factor Maz is essential for normal eye development.
Wnt/β-catenin signaling has an essential role in eye development. Faulty regulation of this pathway results in ocular malformations, owing to defects in cell-fate determination and differentiation. Herein, we show that disruption of Maz, the gene encoding Myc-associated zinc-finger transcription factor, produces developmental eye defects in mice and humans. Expression of key genes involved in the Wnt cascade, Sfrp2, Wnt2b and Fzd4, was significantly increased in mice with targeted inactivation of Maz, resulting in abnormal peripheral eye formation with reduced proliferation of the progenitor cells in the region. Paradoxically, the Wnt reporter TCF-Lef1 displayed a significant downregulation in Maz-deficient eyes. Molecular analysis indicates that Maz is necessary for the activation of the Wnt/β-catenin pathway and participates in the network controlling ciliary margin patterning. Copy-number variations and single-nucleotide variants of MAZ were identified in humans that result in abnormal ocular development. The data support MAZ as a key contributor to the eye comorbidities associated with chromosome 16p11.2 copy-number variants and as a transcriptional regulator of ocular development.


25 years after age-1: genes, interventions and the revolution in aging research.
This communication will briefly review more than 30 years of research on aging using the nematode Caenorhabditis elegans ("The Worm") as carried out in the labs of Tom Johnson. We will highlight research directions initiated in the 1980's, which were exciting for those of us trying to turn over a new leaf in aging research. In this narrative, I will discuss primarily the science that I and my lab have been involved with for the last 30 years. This area has been fascinating to those studying the sociology of science as modern aging research has moved to replace the simplistic, poorly controlled and outright fictitious approaches seen in much of the previous aging research.


ACR-26: a novel nicotinic receptor subunit of parasitic nematodes.
Nematode nicotinic acetylcholine receptors are the targets for many effective anthelmintics, including those recently introduced into the market. We have identified a novel nicotinic receptor subunit sequence, acr-26, that is expressed in all the animal parasitic nematodes we examined from clades III, IV and V, but is not present in the genomes of Trichinella spiralis, Caenorhabditis elegans, Pristionchus pacificus and Meloidogyne spp. In Ascaris suum, ACR-26 is expressed on muscle cells isolated from the head, but not from the mid-body region. Sequence comparisons with other vertebrate and nematode subunits suggested that ACR-26 may be capable of forming a functional homomeric receptor; when acr-26 cRNA was injected into Xenopus oocytes along with Xenopus laevis ric-3 cRNA we occasionally observed the formation of acetylcholine- and nicotine-sensitive channels. The unreliable expression of ACR-26 in vitro may suggest that additional subunits or chaperones may be required for efficient formation of the functional receptors. ACR-26 may represent a novel target for the development of cholinergic anthelmintics specific for animal parasites.


The Use of Chatbots in Oncological Care: A Narrative Review.
Few reports have investigated chatbots in patient care. We aimed to assess the current applications, limitations, and challenges in the literature on chatbots employed in oncological care. We queried the PubMed database through April 2022 and included studies that investigated the use of chatbots in different phases of oncological care. The search used five different combinations of the specific terms "chatbot", "cancer", "oncology", and "conversational agent". Inclusion criteria were chatbot use in any aspect of oncological care-prevention, patient education, treatment, and surveillance. The initial search yielded 196 records, 21 of which met inclusion criteria. The identified chatbots mostly focused on breast and ovarian cancer (n=8), with the second most common being cervical cancer (n=3). Good patient satisfaction was reported among 14 of 21 chatbots. The most reported chatbot applications were cancer screening, prevention, risk stratification, treatment, monitoring, and management. Of 12 studies examining efficacy of care via chatbot, 9 demonstrated improvements compared to standard care. Chatbots used for oncological care to date demonstrate high user satisfaction, and many have shown efficacy in improving patient-centered communication, accessibility to cancer-related information, and access to care. Currently, chatbots are primarily limited by the need for extensive user-testing and iterative improvement before widespread implementation.


Microcavity phonon polaritons from the weak to the ultrastrong phonon-photon coupling regime.
Strong coupling between molecular vibrations and microcavity modes has been demonstrated to modify physical and chemical properties of the molecular material. Here, we study the less explored coupling between lattice vibrations (phonons) and microcavity modes. Embedding thin layers of hexagonal boron nitride (hBN) into classical microcavities, we demonstrate the evolution from weak to ultrastrong phonon-photon coupling when the hBN thickness is increased from a few nanometers to a fully filled cavity. Remarkably, strong coupling is achieved for hBN layers as thin as 10 nm. Further, the ultrastrong coupling in fully filled cavities yields a polariton dispersion matching that of phonon polaritons in bulk hBN, highlighting that the maximum light-matter coupling in microcavities is limited to the coupling strength between photons and the bulk material. Tunable cavity phonon polaritons could become a versatile platform for studying how the coupling strength between photons and phonons may modify the properties of polar crystals.


Agglutination of mouse erythrocytes by binding of non-choline phospholipids to a 70 000-Dalton protein.
The structure of some phospholipids that cause agglutination of mouse erythrocytes has been studied. Haemagglutination is a property of non-choline-containing phospholipids; the phosphate group is essential and unsaturated fatty acids optimal. A protein of Mr 70 000 was isolated from mouse erythrocyte membranes which completely inhibited phospholipid-mediated erythrocyte agglutination. It is proposed that this protein is the phospholipid binding site on mouse erythrocytes and the ligand for the human B-lymphocyte receptor for mouse erythrocytes. Preliminary investigations suggest that a similar inhibitor of phospholipid-mediated agglutination is found in serum. Agglutination of mouse erythrocytes by phospholipid and specific inhibition by the 70 kDa membrane protein constitute a simple system for studying the interaction of phospholipid with protein.


Mantel-Haenszel-type tests for testing equivalence or more than equivalence in comparative clinical trials.
This paper proposes Mantel-Haenszel-type statistics for testing whether a new treatment is at least as effective as the standard treatment in comparative binomial trials. The null hypotheses considered are of a specified nonzero difference and of a ratio not equal to unity. It is shown that we may also use these tests for testing the equivalence of two treatments.


Proteomic analysis of hypothalamus in prepubertal and pubertal female goat.
The functions of proteins at the onset of puberty in goats remain largely unexplored. To identify the proteins regulating puberty in goats, we analysed protein abundance and pathways in the hypothalamus of female goats. We applied tandem mass tag (TMT) labelling, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and parallel reaction monitoring (PRM) to examine hypothalamus of pubertal (cases; n = 3) and prepubertal (controls; n = 3) goats. We identified 5119 proteins, including 69 differentially abundant proteins (DAPs), of which 35 were upregulated and 34 were downregulated. Fourteen DAPs were randomly selected to verify these results using PRM, and the results were consistent with the TMT quantitative results. DAPs were enriched in MAPK signalling pathway, Ras signalling pathway, Autophagy-animal, Endocytosis, and PI3K/Akt/mTOR signalling pathway categories. These pathways are related to embryogenesis, cell proliferation, cell differentiation, and promoting the release of gonadotropin-releasing hormone (GnRH) in the hypothalamus. In particular, PDGFRβ and MAP3K7 occupied important locations in the protein-protein interaction network. The results demonstrate that DAPs and their related signalling pathways are crucial in regulating puberty in goats. However, further research is needed to explore the functions of DAPs and their pathways to provide new insights into the mechanism of puberty onset. SIGNIFICANCE: In domestic animals, reaching the age of puberty is an event that contributes significantly to lifetime reproductive potential. And the hypothalamus functions directly in the complex systemic changes that control puberty. Our study was the first TMT proteomics analysis on hypothalamus tissues of pubertal goats, which revealed the changes of protein and pathways that are related to the onset of puberty. We identified 69 DAPs, which were enriched in the MAPK signaling pathway, the Ras signaling pathway, and the IGF-1/PI3K/Akt/mTOR pathway, suggesting that these processes were probably involved in the onset of puberty.


PRMT1 methylation of WTAP promotes multiple myeloma tumorigenesis by activating oxidative phosphorylation via m6A modification of NDUFS6.
Epigenetic modifications play important roles during the pathogenesis of multiple myeloma (MM). Herein, we found that protein arginine methyltransferase 1 (PRMT1) was highly expressed in MM patients, which was positively correlated with MM stages. High PRMT1 expression was correlated with adverse prognosis in MM patients. We further showed that silencing PRMT1 inhibited MM proliferation and tumorigenesis in vitro and in vivo. Mechanistically, we revealed that the knockdown of PRMT1 reduced the oxidative phosphorylation (OXPHOS) of MM cells through NDUFS6 downregulation. Meanwhile, we identified that WTAP, a key component of the m6A methyltransferase complex, was methylated by PRMT1, and NDUFS6 was identified as a bona fide m6A target of WTAP. Finally, we found that the combination of PRMT1 inhibitor and bortezomib synergistically inhibited MM progression. Collectively, our results demonstrate that PRMT1 plays a crucial role during MM tumorigenesis and suggeste that PRMT1 could be a potential therapeutic target in MM.


Influence of Psoroptes ovis on the energy metabolism of heifer calves.
To determine the effect of Psoroptes ovis on the energy metabolism of heifers, 32 calves were randomly assigned to four treatments in a 2 X 2 factorial arrangement. Calves were fed the same diet at two intake levels, high or low, and were either infested or were not infested with P. ovis mites. Calves were housed in antigrooming stanchions. Body composition was determined by urea dilution on Days 0 and 63. Ration digestibility was determined on fecal grab samples using acid-insoluble ash as a marker. Infested calves had developed a severe P. ovis infestation 7 weeks following exposure and had significantly lower daily gain, gain:feed, and energy retention and higher serum glutamic-oxaloacetic transaminase than control calves. P. ovis infestation increased the maintenance energy requirement of calves by greater than 50% (79 vs. 123 kcal of net energy kg-1 body wt.75). For each 10% increase in the body surface affected by P. ovis, maintenance energy requirement increased 0.5 mcal day-1.


Efficient Removal of Heavy Metals from Aqueous Solutions through Functionalized γ-Graphyne-1 Membranes under External Uniform Electric Fields: Insights from Molecular Dynamics Simulations.
Carbon-based nanosheet membranes with functionalized pores have great potential as water treatment membranes. In this study, the separation of Hg2+ and Cu2+ as heavy metal ions from aqueous solutions using a functionalized γ-graphyne-1 nanosheet membrane is investigated by molecular dynamics simulations. The simulation systems consist of a γ-graphyne-1 nanosheet with -COOH or -NH2 functional groups on the edge of pores placed in an aqueous solution containing CuCl2 and HgCl2. An external electric field is applied as a driving force across the membrane for the separation of heavy metal ions using these functionalized pores. The ion-membrane and water molecule-membrane interaction energies, the radial distribution function of cations, the retention time and permeation of ions through the membrane, the density profile of water and ions, and the hydrogen bond in the system are investigated, and these results reveal that the performance of -NH2-functionalized γ-graphyne-1 is better than that of -COOH-functionalized γ-graphyne-1 in the separation of Cu2+, while the Hg2+ cations encounter a high energy barrier as they pass through the membrane, especially in the -COOH-functionalized pore, due to their larger ionic radius and the smaller pore size of this membrane.


Psychosocial predictors of health status revisited.
A one-year prospective design was used to explore health care costs, claims, and job burnout in relation to the following predictors: work and life stress, social support, exercise, Type A behavior, and personal hardiness. Nowack's (1991) model guided the study. A sample of 260 working adults from two organizations participated. Bivariate correlational results suggested that stressful work events were positively related to health care costs and the number of claims filed. With regard to job burnout, (a) stress appeared positively related to personal accomplishment and (b) Type A behavior was associated with greater emotional exhaustion. Higher job satisfaction, social support, and Type A behavior were associated with an increased sense of personal accomplishment. Findings concerning objective health indicators are discussed in relation to previous results pertaining to self-reports of health/illness.


The metabolic response of the canine neonate to twenty-four hours of fasting.
The metabolic effects of 24 hours of neonatal fasting in unanesthetized dogs were compared to fasting for three hours during the first day of life. Blood glucose, lactate, and ketones were unaltered while FFA (0.94 +/- 0.07 v 0.70 +/- 0.04 mmol/L, P less than .01), glycerol (0.21 +/- 0.01 v 0.12 +/- 0.01 mmol/L, P less than .01), and triglycerides (0.41 +/- 0.03 v 0.23 +/- 0.03 mmol/L, P less than .01) were lower at 24 hours. Glucose production and lactate and alanine turnover were unaffected while palmitate turnover declined (8.8 +/- 0.7 v 5.1 +/- 0.5 mumol/kg/min, P less than .01). Oxygen consumption decreased (6.9 +/- 0.4 v 6.0 +/- 0.3 mL/kg/min, P less than .02) while RQ increased (0.79 +/- 0.02 v 0.86 +/- 0.03, P less than 0.05) at 24 hours. Hepatic glycogen content declined (575 +/- 37 to 266 +/- 32 mumol/g, P less than .001) and could account for a GP of 12 mumol/kg/min between 3 and 24 hours of age. Both gluconeogenesis from lactate and alanine increased, together accounting for 7% and 21% of glucose production at 3 and 24 hours. The increment in gluconeogenesis may be facilitated by augmented hepatic cytosolic phosphoenolpyruvate carboxykinase at 24 hours (1.8 +/- 0.2 v 14.1 +/- 0.8 nmol/min mg protein, P less than .01). Despite the decline in VO2, hepatic ATP and energy charge were unaltered by 24 hours of fasting. These data suggest that FFA availability diminishes during a prolonged neonatal canine fast resulting in lower VO2. Furthermore, as FFA availability declines, glucose utilization becomes the predominant precursor for energy production.(ABSTRACT TRUNCATED AT 250 WORDS)


Treatment with spexin mitigates diet-induced hepatic steatosis in vivo and in vitro through activation of galanin receptor 2.
It was reported that spexin as an adipocyte-secreted protein could regulate obesity and insulin resistance. However, the specific metabolic contribution of spexin to fatty liver remains incompletely understood. Herein, we investigated the effects of spexin on hepatosteatosis and explored the underlying molecular mechanisms. HFD-fed mice were injected with spexin and/or GALR2 antagonist M871, while PA-induced HepG2 cells were treated with spexin in the absence or presence of M871 for 12 h, respectively. Gene expression in liver tissues and hepatocytes was assessed by qRT-PCR and western blotting, respectively. The results showed that body weight, visceral fat content, liver lipid droplet formation, hepatic intracellular triglyceride, and serum triglyceride were reduced in spexin-treated mice. Furthermore, spexin increased the expression of hepatic CPT1A, PPARα, SIRT1, KLF9, PGC-1α and PEPCK in vivo and in vitro. Additionally, spexin treatment improved glucose tolerance and insulin sensitivity in mice fed the HFD. Interestingly, these spexin-mediated beneficial effects were abolished by the GALR2 antagonist M871 in mice fed HFD and PA-induced HepG2 cells, suggesting that spexin mitigated HFD-induced hepatic steatosis by activating the GALR2, thereby increasing CPT1A, PPARα, SIRT1, KLF9, PGC-1α and PEPCK expression. Taken together, these data suggest that spexin ameliorates NAFLD by improving lipolysis and fatty acid oxidation via activation of GALR2 signaling.


Coreceptor reversal in the thymus: signaled CD4+8+ thymocytes initially terminate CD8 transcription even when differentiating into CD8+ T cells.
A central paradigm of T cell development is that CD4+8+ (DP) thymocytes differentiate into CD4+ or CD8+ T cells in response to intrathymic signals that extinguish transcription of the inappropriate coreceptor molecule. Contrary to this prevailing paradigm, we now demonstrate that signaled DP thymocytes initially terminate CD8 transcription even when differentiating into CD8+ T cells. Remarkably, thymocytes that have selectively terminated CD8 transcription can be signaled by IL-7 to differentiate into CD8+ T cells by silencing CD4 transcription and reinitiating CD8 transcription, events we refer to as "coreceptor reversal." These observations significantly alter our understanding of CD8+ T cell differentiation and lead to a new perspective ("kinetic signaling") on CD4/CD8 lineage determination in the thymus. These observations also suggest a novel mechanism by which bipotential cells throughout development can determine their appropriate cell fate.


An improved polymerase chain reaction method for genetic testing of spinocerebellar ataxia type 3.
The development of a reliable PCR assay for genetic testing of spinocerebellar ataxia type 3. Touchdown PCR conditions were tested and different primer sets were evaluated with genomic DNA from blood sample of patients suffering from spinocerebellar ataxia type 3 (SCA3). An improved PCR assay was developed with a new set of primers and using the optimized touchdown PCR protocol. This new assay had been successfully employed in the screening of one identificated SCA3 family. Results from the present study document a simple and reliable PCR assay for genetic testing of SCA3. Strategies used in the present study may find applications in the optimization of PCR assay for triplet expansion with GC rich in the sequence context.


Effects of lysophospholipids on tumor microenvironment.
The effects of lysophospholipids (LPLs) on cancer microenvironment is a vast and growing field. These lipids are secreted physiologically by various cell types. They play highly important roles in the development, activation and regulation of the immune system. They are also secreted by cancerous cells and there is a strong association between LPLs and cancer. It is clear that these lipids and in particular sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) play major roles in regulating the growth of tumor cells, and in manipulating the immune system. These activities can be divided into two parts; the first involves the ability of S1P and LPA to either directly or through some of the enzymes that generate them such as sphingosine kinases or phospholipases, induce the motility and invasiveness of tumor cells. The second mechanism involves the recently discovered effects of these lipids on the anti-tumor effector natural killer (NK) cells. Whereas S1P and LPA induce the recruitment of these effector cells, they also inhibit their cytolysis of tumor cells. This may support the environment of cancer and the ability of cancer cells to grow, spread and metastasize. Consequently, LPLs or their receptors may be attractive targets for developing drugs in the treatment of cancer where LPLs or their receptors are up-regulated.


Penalization in the treatment of amblyopia.
Penalization is a useful alternative to occlusion therapy in amblyopia of mild and moderate degrees and in preserving therapeutic gains made by previous occlusion treatment. Penalization at near, total, and alternating penalization are more valuable than penalization at distance. Because penalization does not prevent the development of visual deprivation amblyopia, unilateral atropine therapy must be used carefully in treating patients in an age group susceptible to occlusion amblyopia.


Evaluation of groundwater quality and human health risks from fluoride and nitrate in semi-arid region of northern India.
Groundwater quality in the alluvial plains of Punjab has special significance and needs great attention since it is the foremost source of drinking, irrigation and industrial uses. The present research work emphasizes the integrated hydrogeochemical and chemometric statistical approaches to appraise the geochemical processes and source apportionment of the groundwater in the alluvial plains of Jalandhar district, Punjab, India. The human health risk assessment was also performed to quantify the potential non-carcinogenic impacts of nitrate and fluoride on human health through ingestion of groundwater. For this purpose, 41 groundwater samples were collected from different groundwater abstraction units and analysed for pH, electrical conductivity, total dissolved solids, total hardness, total alkalinity and major ions (Ca2+, Mg2+, Na+, K+, HCO3-, CO32-, SO42-, NO3-, F-, Cl- and PO43-) using standard protocols. Drinking water quality index and Revelle index showed that groundwater samples fall under poor to unfit water class and salinization along the south-western portion of the study region shows poor water quality. The results of the hazard index (HIingestion) show 68% and 46.34% of the groundwater samples have HI > 1 for children and adults. The non-carcinogenic health risk assessment of nitrate (NO3-) and fluoride (F-) on the local population indicated that the children are more vulnerable through direct ingestion of drinking water than adults. Piper diagram and saturation index reveal that Ca2+-Mg2+-HCO3- is the dominant hydrochemical facies and oversaturated with calcite, dolomite and aragonite minerals in the groundwater. Gibbs diagrams, chloro-alkaline indices and scatter plots show that the hydrochemistry of the groundwater is mainly governed by aquifer material interaction such as weathering of silicate, carbonate rock, halite dissolution and cation exchange process. Chemometric statistical techniques revealed that the source identification of parameters such as Ca2+, Mg2+, Na+, K+, HCO3-, CO3- and F- is originated from geogenic factors, whereas NO3-, SO42-, Cl- and PO43- are from the anthropogenic origin. Therefore, urgent and efficient measures must be taken to combat groundwater pollution and reduce human health risk in the study area.


The acceptability of five periapical radiographic techniques to dentists and patients.
Dentists are now advised to use film holders as an aid to intra-oral radiography. The aim of this investigation was to compare a range of commercially available film holders for periapical radiography with the traditional bisecting angle technique using the patient's finger as support for the film, in terms of the ease of use for the dentist and patient comfort. Twelve general dental practitioners (GDPs) were asked to perform 50 periapical examinations using their conventional (bisecting angle) technique (BA) followed by 50 using, in turn, four film holders: the Stabe (S), the Eggen (E), the Rinn (R) and the 'Superbite' devices (SB). There was no difference between BA, S and SB techniques in terms of difficulty experienced by the GDPs. However, both E and R film holders led to an increase in examinations reported as 'difficult' to perform. There was no difference in patient discomfort between the BA and SB techniques, but the use of the E and R film holders and, to a lesser degree, the S film holder was associated with increased discomfort. There was no consistent association between dentists' assessments of examinations as 'difficult to perform' and patients' opinions on discomfort. It is concluded that SB film holders provide a simple means of intraoral radiographic positioning with no increase in patient discomfort.


Psychological distress, substance use, and adjustment among parents living with HIV.
Being a parent, especially a custodial parent, living with HIV was anticipated to increase psychological distress and challenges to self-care. Mental health symptoms, substance use, and health care utilization were assessed among 3818 HIV-infected adults, including custodial parents, noncustodial parents, and nonparents, in 4 AIDS epicenters. Custodial parents demonstrated significantly poorer medication adherence and attendance at medical appointments but were similar to nonparents and noncustodial parents in mental health symptoms and treatment utilization for mental health and substance use problems. Noncustodial parents demonstrated the highest levels of recent substance use and substance abuse treatment. Other markers of risk, such as African American ethnicity, lack of current employment income, and injection drug use moderated many of the apparent psychosocial disadvantages exhibited by parents. Interventions specific to the psychosocial stressors facing families living with HIV are needed.


Inhibition of interaction between ROCK1 and Rubicon restores autophagy in endothelial cells and attenuates brain injury after prolonged ischemia.
Acute ischemic stroke (AIS) induces cerebral endothelial cell death resulting in the breakdown of the blood-brain barrier (BBB). Endothelial cell autophagy acts as a protective mechanism against cell death. Autophagy is activated in the very early stages of ischemic stroke and declines after prolonged ischemia. Previous studies have shown that Rubicon can inhibit autophagy. The current study aimed to investigate whether continuous long-term ischemia can inhibit autophagy in endothelial cells after ischemic stroke by regulating the function of Rubicon and its underlying mechanism. Wild-type male C57BL/6J mice were subjected to transient middle cerebral artery occlusion (tMCAO). ROCK1, ROCK2, and NOX2 inhibitors were injected into male mice 1 h before the onset of tMCAO. Disease severity and BBB permeability were evaluated. bEnd.3 cells were cultured in vitro and subjected to oxygen-glucose deprivation (OGD). bEnd.3 cells were pretreated with or without ROCK1, ROCK2, or NOX2 inhibitors overnight and then subjected to OGD. Cell viability and permeability were also evaluated. The expression of Rubicon, ROCK1, and autophagy-related proteins were analyzed. Increased BBB permeability was correlated with Rubicon expression in tMCAO mice and Rubicon was upregulated in endothelial cells subjected to OGD. Autophagy was inhibited in endothelial cells after long-term OGD treatment and knockdown of Rubicon expression restored autophagy and viability in endothelial cells subjected to 6-h OGD. ROCK1 inhibition decreased the interaction between Beclin1 and Rubicon and restored cell viability and autophagy suppressed by 6-h OGD treatment in endothelial cells. Additionally, ROCK1 inhibition suppressed Rubicon, attenuated BBB disruption, and brain injury induced by prolonged ischemia in 6-h tMCAO mice. Prolonged ischemia induced the death of brain endothelial cells and the breakdown of the BBB, thus aggravating brain injury by increasing the interaction of ROCK1 and Rubicon with Beclin1 while inhibiting canonical autophagy. Inhibition of ROCK1 signaling in endothelial cells could be a promising therapeutic strategy to prolong the therapeutic time window in AIS.


Anorexia nervosa patients display a deficit in membrane long chain poly-unsaturated fatty acids.
The energy deficit in anorexia nervosa (AN) results from a low energy and fat intake. Data from the literature are at odds about a deficiency in essential fatty acids (EFA), linoleic (LA) and alpha-linolenic (ALA) acid. The aim of the present study was to determine RBC phospholipid FA composition in AN patients, and to relate this FA profile to fat mass and BMI. We postulated that there should not be a deficit in EFA, because AN patients are in negative energy balance, and a significant supply of EFA arises from the lipolysis of AT triglycerides. Twenty-two AN women (14.2 ± 2.0 kg/m² BMI) had a DEXA-body composition analysis, and an analysis of FA composition in the phospholipids of red blood cell membranes by capillary gas chromatography. The control group was made of 25 healthy women. There was no EFA deficiency in RBC, i.e. no decrease in LA and ALA, and no rise in C20:3 n-9. However, long chain poly-unsaturated FA (LC-PUFA), mainly those of the n-3 family were significantly reduced. A negative correlation was found between fat mass and ALA, with a critical fat mass close to 5 kg, under which ALA was dramatically increased. AN patients display a complex membrane FA profile without traits of EFA deficiency but with a deficiency in LC-PUFA. We propose that adipose tissue is an endogenous source of FA able to compensate for the reduced food intake in EFA, but unable to provide enough LC-PUFA.


Efficacy of 5% imiquimod for the treatment of Vaginal intraepithelial neoplasia-A systematic review of the literature and a meta-analysis.
Vaginal intraepithelial neoplasia (VaIN) is an uncommon disease associated with HPV and is considered to be a precursor of vaginal carcinoma. To date, treatment recommendations vary with no universally accepted standard of care as best treatment modality. Nevertheless, 5% imiquimod appears to be a promising, alternative, non-invasive treatment option. To ascertain the efficacy of 5% imiquimod for the treatment of this rare condition, we conducted a systematic review and meta-analysis of the proportion of women who received 5% imiquimod with their complete response, HPV clearance and recurrence rates. A literature search was carried out throughout the PubMed, EMBASE, ClinicalTrials.gov and Cochrane Databases for relevant studies. We computed the summary proportions for complete response, HPV clearance and non-recurrence following administration of 5% imiquimod by random effects meta-analysis. Six articles reporting on 94 patients were included. The summary proportions of women with complete response and HPV clearance were 76.5% (95% CI 59.4-98.5) and 52.5% (95% CI 29.5-93.6) respectively. The summary proportion of women with non-recurrence appeared high (94.3% (95% CI 67.1-132)), yet not significant. Use of 5% imiquimod for the treatment of VaIN is associated with relatively high response rate, satisfactory HPV clearance, whilst the risk for VaIN recurrence appears low.


Involvement of autophagy in the maintenance of rat intervertebral disc homeostasis: an in-vitro and in-vivo RNA interference study of Atg5.
In the largest avascular low-nutrient intervertebral disc, resident cells would utilize autophagy, a stress-response survival mechanism by self-digestion and recycling wastes. Our goal was to elucidate the involvement of autophagy in disc homeostasis through RNA interference of autophagy-related gene 5 (Atg5). In vitro, small interfering RNAs (siRNAs) targeting autophagy-essential Atg5 were transfected into rat disc cells. Cell viability with levels of autophagy including Atg5 expression, apoptosis, and senescence was assessed under serum starvation and/or pro-inflammatory interleukin-1 beta (IL-1β) stimulation. In vivo, time-course autophagic flux was monitored following Alexa Fluor® 555-labeled Atg5-siRNA injection into rat tail discs. Furthermore, 24-h temporary static compression-induced disruption of Atg5 siRNA-injected discs was observed by radiography, histomorphology, and immunofluorescence. In disc cells, three different Atg5 siRNAs consistently suppressed autophagy with Atg5 protein knockdown (mean 44.4% [95% confidence interval: -51.7, -37.1], 51.5% [-80.5, -22.5], 62.3% [-96.6, -28.2]). Then, Atg5 knockdown reduced cell viability through apoptosis and senescence not in serum-supplemented medium (93.6% [-0.8, 21.4]) but in serum-deprived medium (66.4% [-29.8, -8.6]) further with IL-1β (44.5% [-36.9, -23.5]). In disc tissues, immunofluorescence detected intradiscal signals for the labeled siRNA even at 56-d post-injection. Immunoblotting found 56-d autophagy suppression with prolonged Atg5 knockdown (33.2% [-52.8, -5.3]). With compression, Atg5 siRNA-injected discs presented radiographic height loss ([-43.9, -0.8]), histological damage ([-5.5, -0.2]), and immunofluorescent apoptosis ([2.2, 22.2]) and senescence ([4.1, 19.9]) induction compared to control siRNA-injected discs at 56 d. This loss-of-function study suggests Atg5-dependent autophagy-mediated anti-apoptosis and anti-senescence. Autophagy could be a molecular therapeutic target for degenerative disc disease.


Cell specific effect of ecdysone on RNA synthesis in the differentiated salivary gland of Acricotopus lucidus.
The influence of the steroid hormone ecdysone on the RNA metabolism of 2 cell types (main and anterior lobe) of the Acricotopus salivary gland has been investigated. Ecdysone has been found to increase the synthesis of 4S RNA and poly(A)+ RNA in the main lobe, while the anterior lobe remained unaffected by the same treatement. The stimulated poly(A)+ RNA is characterized by its relatively low cytoplasmic turnover. No evidence has been found for the stimulation of ribosomal RNA synthesis in both cell types. Analysis of the metabolic stability of cytoplasmic poly(A)+ RNA after ecdysone administration revealed a different response of the main and anterior lobe cells. Only in the anterior lobe ecdysone causes a shortening of the lifetime of that part of poly(A)+ RNA which is peculiar in its early appearance and relatively low stability in the cytoplasm.


Pelvic sonography and uterine artery color Doppler analysis in the diagnosis of female precocious puberty.
To evaluate the role of ultrasound and color Doppler analyses in the diagnosis of precocious puberty. Gray-scale sonographic uterine and ovarian evaluation together with color Doppler analysis of the uterine artery were prospectively performed in 29 girls presenting with premature breast development and pubic hair growth. The values were compared with results obtained from the gonodotrophin releasing hormone stimulation test. Excluded from the study were patients with isolated thelarche or isolated pubarche and those patients with gonodotrophin releasing hormone-independent puberty and with polycystic ovaries. According to the Tanner scale, all the girls presented a breast stage of 2-3 and pubic hair stage 2-3. The uterine size was greater in those girls who presented a pubertal response to the gonodotrophin releasing hormone test (Group II; n = 20) (8.07 +/- 4.47 mL) than in those who did not (Group I; n = 9) (3.07 +/- 1.18 mL; P = 0.001). The ovarian volume and the number of small follicles was not significantly different between the groups. On Doppler analysis, more elevated impedances were observed in Group I (pulsatility index = 3.28 +/- 0.37) than in Group II (pulsatility index = 2.29 +/- 0.19; P = 0.001) girls. The presence of a low pulsatility index (< 2.5) at the level of the uterine arteries had a high diagnostic value for precocious puberty (sensitivity 86%, specificity 100%). Uterine artery Doppler analysis may assist the diagnosis of gonodotrophin releasing hormone-dependent precocious puberty, may be useful for the selection of those girls needing treatment, and may simplify the follow-up of girls treated for precocities.


Paracrine factors in adult rat testis gonadotrophin control of opioids and LHRH like peptide.
A paracrine regulation involves agents which are produced by one cell type and act on an other one within an organ. In rodent testis, local control mechanisms modulate the actions of the gonadotrophins according to local requirements. Two groups of peptides-opioids and testicular LHRH are defined as paracrine factors and in vivo they are both modified by HCG. In vitro, after HCG exposure, we first localized an opioid like material in Sertoli cells cytoplasma by immunohistochemistry. This material is detected in freeze dried homologous culture media using a dot immunobinding technique. With a longer HCG exposure, an LHRH like material is then visualized in the basal compartment of the Sertoli cells and it is detected in freeze dried homologous culture media by the same technical procedure than for opioid material. By adding synthetic enkephalins to culture medium, we obtain the same results as with the endogenous opioid material, excreted after HCG addition. If naloxone a potent opiate antagonist, is added to the culture medium previously to HCG or enkephalins, the Sertoli cells cytoplasma are no more immunoreactives with the anti-enkephalin serum and no LHRH material is neither visualized by immunohistochemical technique neither detected in culture media. We conclude that testicular opioids, synthetized by the Leydig cells and which have specific Sertoli cells receptors are one Leydig-Sertoli paracrine communication factor. One way of response to their receptor fixation is the synthesis and excretion by Sertoli cells of testicular LHRH. This one is known to act on Leydig cells via specific receptors and it is one Sertoli-Leydig cells paracrine communication factor.(ABSTRACT TRUNCATED AT 250 WORDS)


Doctoral education from a distance.
This article describes the environmental factors that have contributed to the recent rapid growth of nursing doctoral education at a distance. Early and recent efforts to deliver distance doctoral education are discussed, using The University of Arizona College of Nursing experience as the key exemplar. The Community of Inquiry model is introduced as an appropriate model for doctoral education and then used as a framework to evaluate the current state of the art in distance doctoral nursing education. Successes and challenges in delivering doctoral education from a distance are described.


Kinase regulation: competing phosphatases in JAK dephosphorylation.
The Janus kinases (JAKs) are essential for cytokine receptor signalling and their dephosphorylation represents a potent inhibitory mechanism. A new paper highlights the T-cell tyrosine phosphatase as an important JAK inhibitor.


Comparison of commonly used antimicrobial susceptibility testing methods for evaluating susceptibilities of clinical isolates of Enterobacteriaceae and nonfermentative Gram-negative bacilli to cefoperazone-sulbactam.
The aim of this study was to investigate the cefoperazone-sulbactam (CFP-SUL) susceptibilities of important Gram-negative bacteria (GNB) by agar dilution (reference method), disk diffusion, and two automated methods. A total of 799 GNB isolates, including Enterobacteriaceae (n = 500) and nonfermentative GNB (NFGNB, n = 299), were recovered from various clinical specimens collected at National Taiwan University Hospital, Taipei, Taiwan from November 2013 to December 2014. The agar dilution method, disk diffusion method, and two automated susceptibility systems (Phoenix and Vitek 2) were used for testing susceptibility of the isolates to CFP-SUL. Categories of susceptibility (susceptible, intermediate, or resistant) to CFP-SUL yielded from each method were interpreted according to CFP-SUL interpretive breakpoints proposed previously. The results of categorical agreement and errors obtained between the agar dilution method and the other three methods were analyzed. The Vitek 2 system had the highest error rates against Escherichia coli (n = 150) and Enterobacter cloacae (n = 77) isolates, i.e., 6.7% and 11.7% minor errors, 8.5% and 1.7% major errors, and 40% and 20% very major errors, respectively. Additionally, the Vitek 2 system was also found to have a significantly lower sensitivity (44.4%) and lower positive predictive value (18.2%) for detecting CFP-SUL nonsusceptible E. coli isolates than other methods. For carbapenem-nonsusceptible Enterobacteriaceae isolates, the Vitek 2 system failed to detect correct susceptibility to CFP-SUL. The three methods failed to correctly detect CFP-SUL susceptibility categories against all NFGNB isolates except Pseudomonas aeruginosa. The Vitek 2 system is a suboptimal method in correctly detecting CFP-SUL susceptibility categories for E. coli, E. cloacae, and carbapenem-nonsusceptible Enterobacteriaceae isolates.


Association of Small Fiber Function with Microvascular Perfusion of Peripheral Nerves in Patients with Type 2 Diabetes : Study using Quantitative Sensory Testing and Magnetic Resonance Neurography.
Diabetic small fiber neuropathy (SFN) is caused by damage to thinly myelinated A‑fibers (δ) and unmyelinated C‑fibers. This study aimed to assess associations between quantitative sensory testing (QST) and parameters of peripheral nerve perfusion obtained from dynamic contrast enhanced (DCE) magnetic resonance neurography (MRN) in type 2 diabetes patients with and without SFN. A total of 18 patients with type 2 diabetes (T2D, 8 with SFN, 10 without SFN) and 10 healthy controls (HC) took part in this cross-sectional single-center study and underwent QST of the right leg and DCE-MRN of the right thigh with subsequent calculation of the sciatic nerve constant of capillary permeability (Ktrans), extravascular extracellular volume fraction (Ve), and plasma volume fraction (Vp). The Ktrans (HC 0.031 min-1 ± 0.009, T2D 0.043 min-1 ± 0.015; p = 0.033) and Ve (HC 1.2% ± 1.5, T2D: 4.1% ± 5.1; p = 0.027) were lower in T2D patients compared to controls. In T2D patients, compound z‑scores of thermal and mechanical detection correlated with Ktrans (r = 0.73; p = 0.001, and r = 0.57; p = 0.018, respectively) and Ve (r = 0.67; p = 0.002, and r = 0.69; p = 0.003, respectively). Compound z‑scores of thermal pain and Vp (r = -0.57; p = 0.015) correlated negatively. The findings suggest that parameters of peripheral nerve microcirculation are related to different symptoms in SFN: A reduced capillary permeability may result in a loss of function related to insufficient nutritional supply, whereas increased capillary permeability may be accompanied by painful symptoms related to a gain of function.


Directing Group-Promoted Inert C-O Bond Activation Using Versatile Boronic Acid as a Coupling Agent.
A simple Ni(cod)2 and carbene mediated strategy facilitates the efficient catalytic cross-coupling of methoxyarenes with a variety of organoboron reagents. Directing groups facilitate the activation of inert C-O bonds in under-utilized aryl methyl ethers enabling their adaptation for C-C cross-coupling reactions as less toxic surrogates to the ubiquitous haloarenes. The method reported enables C-C cross-coupling with readily available and economical arylboronic acid reagents, which is unprecedented, and compares well with other organoboron reagents with similarly high reactivity. Extension to directing group assisted chemo-selective C-O bond cleavage, and further application towards the synthesis of novel bifunctionalized biaryls is reported. Key to the success of this protocol is the use of directing groups proximal to the reaction center to facilitate the activation of the inert C-OMe bond.


The influence of the type of embryo culture media on the outcome of IVF/ICSI cycles.
Embryo culture media are important factors in IVF, which can significantly influence clinical outcome of IVF/ICSI cycles. Despite this, it is still not completely clear which formulation is most optimal and whether sequential or continuous media should be favored. This study retrospectively analyzed the outcome of IVF/ICSI cycles with regard to different types of culture media used to culture embryos, namely sequential and two types of single step continuous embryo culture media. If the data were combined for both types of single step continuous embryo culture media the only significant difference we observed was the proportion of poor quality embryos on day 3, which was significantly higher (16.9% vs. 22.5%; P = 0.017) in the sequential media. The pregnancy (55.1% vs. 40.5%; P = 0.113) and live birth rates (42.9% vs. 33.8%; P = 0.308) were lower in continuous media, although the difference was not statistically significant. Furthermore, the blastocyst rate (sequential vs. continuous; 47.4% vs. 47.3%; P = 1), and birthweight (3280 ± 630g vs. 3272 ± 575g; P = 0.96) did not significantly differ regardless of the medium used to culture embryos. Additional comparison of each type of continuous medium to sequential media revealed that the difference in the quality of cleavage stage embryos for combined data of both continuous culture media may be derived from the group of cycles were SAGE 1-Step was used to culture embryos. These results therefore indicate that continuous media can be equivalent to sequential media and could help lower the workload in busy IVF labs without impairing the clinical results. Although, caution is needed because this study is limited by its retrospective design. To confirm the results, especially in terms of live birth rates and perinatal outcome, a prospective study is needed with a higher number of included couples.


Detecting proton flux across chromatophores driven by F0F1-ATPase using N-(fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt.
N-(Fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (F-DHPE) is a lipid fluorescence dye sensitive to pH changes and is used in this study for detecting proton flux through F0F1-ATPase within chromatophores driven by ATP hydrolysis. F-DHPE is easily labeled to the outer surface of chromatophores. In the range of pH 7.0 to 9.0, fluorescence intensity is sensitive to pH changes. The sensitivity is especially great in the range of pH 8.2 to 9.0, so pH 8.6 was chosen as the appropriate experimental condition. It is shown that added ATP not only acts as a fluorescence quencher but also can be hydrolyzed by F0F1-ATPase to pump protons into chromatophores, resulting in fluorescence restoration. A stimulator (NaSO3) and various types of inhibitors (NaN3, 5'-adenylyl imidodiphosphate [AMP-PNP], and N,N'-dicyclohexylcarbodiimide [DCCD]) of F0F1 confirmed that fluorescence restoration is caused by ATP-driven proton flux. When loaded with one antibody (anti-beta antibody) or two antibodies (anti-beta antibody and sheep to rabbit second antibody), F0F1-ATPase exhibits lower proton pumping activities, as indicated by fluorescence restoration. The possible mechanism of the inhibition of antibodies on proton pumping activity is discussed.


Nop2 is required for mammalian preimplantation development.
Nucleolar protein 2 (NOP2) is evolutionarily conserved from yeast to human, and has been found to play an important role in accelerating cell proliferation, cell-cycle progression, and tumor aggressiveness. The expression pattern and function of Nop2 during early mammalian embryo development, however, has not been investigated. We identified Nop2 as an essential gene for development to the blastocyst stage while performing an RNA interference (RNAi)-based screen in mouse preimplantation embryos. Nop2 is expressed throughout preimplantation development, with highest mRNA and protein accumulation at the 8-cell and morula stages, respectively. RNAi-mediated knockdown of Nop2 results in embryos that arrest as morula. NOP2-deficient embryos exhibit reduced blastomere numbers, greatly increased apoptosis, and impaired cell-lineage specification. Furthermore, knockdown of Nop2 results in global reduction of all RNA species, including rRNA, small nuclear RNA, small nucleolar RNA, and mRNA. Taken together, our results demonstrate that Nop2 is an essential gene for blastocyst formation, and is required for RNA processing and/or stability in vivo during preimplantation embryo development in the mouse.


Implementation of mandatory opioid prescribing limits in North Carolina: healthcare administrator and prescriber perspectives.
Recent increases in state laws to reduce opioid prescribing have demonstrated a need to understand how they are interpreted and implemented in healthcare systems. The purpose of this study was to explore the systems, strategies, and resources that hospital administrators and prescribers used to implement the 2017 North Carolina Strengthen Opioid Prevention (STOP) Act opioid prescribing limits, which limited initial prescriptions to a five (for acute) or seven (for post-surgical) days' supply. We interviewed 14 hospital administrators and 38 prescribers with degrees in medicine, nursing, pharmacy, business administration and public health working across North Carolina. Interview guides, informed by the Consolidated Framework for Implementation Research, explored barriers and facilitators to implementation. Interview topics included communication, resources, and hospital system support. Interviews were recorded and transcribed, then analyzed using flexible coding, integrating inductive and deductive coding, to inform analytic code development and identify themes. We identified three main themes around implementation of STOP act mandated prescribing limits: organizational communication, prescriber education, and changes in the electronic medical record (EMR) systems. Administrators reflected on implementation in the context of raising awareness and providing reminders to facilitate changes in prescriber behavior, operationalized through email and in-person communications as well as dedicated resources to EMR changes. Prescribers noted administrative communications about prescribing limits often focused on legality, suggesting a directive of the organization's policy rather than a passive reminder. Prescribers expressed a desire for more spaces to have their questions answered and resources for patient communications. While hospital administrators viewed compliance with the law as a priority, prescribers reflected on concerns for adequately managing their patients' pain and limited time for clinical care. Hospital administrators and prescribers approached implementation of the STOP act prescribing limits with different mindsets. While administrators were focused on policy compliance, prescribers were focused on their patients' needs. Strategies to implement the mandate then had to balance patient needs with policy compliance. As states continue to legislate to prevent opioid overdose deaths, understanding how laws are implemented by healthcare systems and prescribers will improve their effectiveness through tailoring and maximizing available resources.


The hot clot sign. A new finding in deep venous thrombosis on bone scintigraphy.
Focal Tc-99m MDP uptake on bone scanning in regions of deep venous thrombosis (DVT) is reported in two patients with documented DVT. It is speculated that this uptake may be related to localized calcification or ossification, which is occasionally observed on radiographs of patients with chronic DVT.


Bonding of restorative materials to dentine: the present status in the Federal Republic of Germany.
It is desirable for filling materials to adhere over the entire area of contact with enamel and dentine. At present we try to achieve adhesion to dentine either by intermediate linking layers (primers) or by the use of 'adhesive' filling materials. Bond strength to dentine attainable with the adhesive glass ionomer cements ChemFil II, Fuji-Ionomer Type II and Ketac-Fil and the primers Scotchbond and Dentin-Adhesit were determined. The tests were carried out on specimens stored for 3 and 21 days in distilled water at 37 degrees C; they were also subjected to thermal cycling. The highest measured bond strength with the glass ionomer cements was about 3.2 MN/m2; etching of the dentine did not generally intensify the bond, while thermal cycling had a slightly negative effect on adhesion. The highest bond strength measured with the primers was about 4.7 MN/m2. There were clear differences between the individual products; in those with very strong primary adhesion, etching of the dentine resulted in impairment of the bond, while thermal cycling had a clear adverse effect.


How many trials are required for parameter estimation in diffusion modeling? A comparison of different optimization criteria.
Diffusion models (Ratcliff, 1978) make it possible to identify and separate different cognitive processes underlying responses in binary decision tasks (e.g., the speed of information accumulation vs. the degree of response conservatism). This becomes possible because of the high degree of information utilization involved. Not only mean response times or error rates are used for the parameter estimation, but also the response time distributions of both correct and error responses. In a series of simulation studies, the efficiency and robustness of parameter recovery were compared for models differing in complexity (i.e., in numbers of free parameters) and trial numbers (ranging from 24 to 5,000) using three different optimization criteria (maximum likelihood, Kolmogorov-Smirnov, and chi-square) that are all implemented in the latest version of fast-dm (Voss, Voss, & Lerche, 2015). The results revealed that maximum likelihood is superior for uncontaminated data, but in the presence of fast contaminants, Kolmogorov-Smirnov outperforms the other two methods. For most conditions, chi-square-based parameter estimations lead to less precise results than the other optimization criteria. The performance of the fast-dm methods was compared to the EZ approach (Wagenmakers, van der Maas, & Grasman, 2007) and to a Bayesian implementation (Wiecki, Sofer, & Frank, 2013). Recommendations for trial numbers are derived from the results for models of different complexities. Interestingly, under certain conditions even small numbers of trials (N < 100) are sufficient for robust parameter estimation.


Comparison of voiding parameters in men and women with lower urinary tract symptoms.
The aim of this study was to compare voiding parameters by using urodynamic investigations in men and women with lower urinary tract symptoms (LUTS). A total of 164 individuals (76 men and 88 women) completed the International Prostate Symptom Score (IPSS) and underwent a detailed urodynamic investigation. Patients were stratified by voiding function, which included bladder voiding efficiency (BVE) of < 80% and of >or= 80%. IPSSs were similar in the men and women except for voiding symptoms for those with a BVE of >or= 80% (12 for men vs. 9 for women, P=0.016). When voiding parameters were compared according to BVE in men and women, respectively, maximum flow rate (Q(max)) (P=0.002), average flow rate (Q(ave)) (P=0.003), voided volume (P=0.037), post-void residual (PVR) (P<0.001), and bladder outlet obstruction index (P=0.016) were different for men with a BVE of < 80% and men with a BVE of >or= 80%. However, for women with a BVE of < 80% or >or= 80%, PVR (P<0.001), detrusor pressure at maximum flow rate (P=0.022), and opening detrusor pressure (P=0.024) were different. The voiding parameters of women differ from those of men according to voiding function. The present results show that a pressure-flow study may represent a difference according to BVE in women as well as in men. Our findings suggest that opening detrusor pressure in women reflects the urethral and detrusor during voiding phase accurately as detrusor pressure at maximum flow rate does.