Thrombospondin 1 activates the macrophage Toll-like receptor 4 pathway.
Previously, we demonstrated that macrophages from thrombospondin 1 (TSP1)-deficient mice have a reduced inflammatory phenotype, suggesting that TSP1 plays a role in macrophage activation. In this study, we determined how TSP1 regulates macrophage function. We found that recombinant or purified platelet human TSP1 treatment stimulated tumor-necrosis factor (TNF)-α expression in bone marrow-derived macrophages in a time- and dose-dependent manner. Toll-like receptor 4 (TLR4) expression (at the mRNA and protein levels) and nuclear factor-kappaB (NF-κB) activity were also stimulated by TSP1 treatment. The TSP1-mediated increase in TNF-α production was abolished in TLR4-deficient macrophages, suggesting that TSP1 activates macrophages through a TLR4-dependent pathway. TSP1 also stimulated TLR4 activation in macrophages in vivo. Furthermore, TSP1-mediated macrophage activation was attenuated by using a peptide or an antibody to block the association between TSP1 and CD36. Taken together, these data suggest that the stimulation of the macrophage TLR4 pathway by TSP1 is partially mediated by the interaction of TSP1 with its receptor, CD36.


Effect of fundoplication on the gastric emptying of liquids.
To evaluate the effect of fundoplication on the gastric emptying (GE) of liquids, the authors studied 96 male Wistar rats divided into three main groups: group E (early postoperative), formed by 32 rats that received physiological saline as a test meal and whose gastric emptying was evaluated 8 days after surgery; group L (late postoperative), which received the same test meal but was evaluated 29 days after surgery; and group G (glucose), which received 5% glucose in water and was studied 8 days after surgery. Each group was subdivided in two subgroups of 16 animals: in one (atropine), the animals received intravenous (I.V.) atropine sulfate (0.3 mg/100 mg rat weight) 60 minutes before GE test; the other subgroup (controls) received I.V. physiological saline. In both subgroups 8 animals had been submitted to fundoplication and 8 to sham operation. Every test meal, containing 6 mg% red phenol, was infused by gravity through a metallic catheter. Gastric retention was determined by measuring the concentration of the marker in the liquid recovered from the stomach 10 minutes after infusion. In the animals of group E, fundoplication increased the gastric emptying of physiological saline, both in the control and the atropine subgroups. In the L group, gastric retention values were similar in fundoplication and sham-operated rats, suggesting an adaptation of the stomach to the fundoplication. In the G group, fundoplication enhanced GE among the control animals, but not among those receiving I.V. atropine sulfate. These results support the importance of gastric emptying studies in every patient to be submitted to fundoplication.


Cyclic delamination behavior of plasma-sprayed hydroxyapatite coating on Ti-6Al-4V substrates in simulated body fluid.
This study aimed to clarify the effect of a simulated body fluid (SBF) on the cyclic delamination behavior of a plasma-sprayed hydroxapatite (HAp) coating. A HAp coating is deposited on the surfaces of surgical metallic materials in order to enhance the bond between human bone and such surfaces. However, the HAp coating is susceptible to delamination by cyclic loading from the patient's gait. Although hip joints are subjected to both positive and negative moments, only the effects of tensile bending stresses on vertical crack propagation behavior have been investigated. Thus, the cyclic delamination behavior of a HAp coating was observed at the stress ratio R=-1 in order to determine the effects of tensile/compressive loading on the delamination behavior. The delamination growth rate increased with SBF immersion, which decreased the delamination life. Raman spectroscopy analysis revealed that the selective phase dissolution in the HAp coating was promoted at interfaces. Finite element analysis revealed that the energy release rate Gmax showed a positive value even in cases with compressive loading, which is a driving force for the delamination of a HAp coating. A prediction model for the delamination growth life was developed that combines a fracture mechanics parameter with the assumed stress-dependent dissolution rate. The predicted delamination life matched the experimental data well in cases of lower stress amplitudes with SBF.


Safety and Immunogenicity of the ID93 + GLA-SE Tuberculosis Vaccine in BCG-Vaccinated Healthy Adults: A Randomized, Double-Blind, Placebo-Controlled Phase 2 Trial.
This randomized, double-blind, placebo-controlled, phase 2a trial was conducted to evaluate the safety and immunogenicity of the ID93 + glucopyranosyl lipid adjuvant (GLA)-stable emulsion (SE) vaccine in human immunodeficiency virus (HIV)-negative, previously Bacillus Calmette-Guérin (BCG)-vaccinated, and QuantiFERON-TB-negative healthy adults in South Korea. Adults (n = 107) with no signs or symptoms of tuberculosis were randomly assigned to receive three intramuscular injections of 2 μg ID93 + 5 μg GLA-SE, 10 μg ID93 + 5 μg GLA-SE, or 0.9% normal saline placebo on days 0, 28, and 56. For safety assessment, data on solicited adverse events (AEs), unsolicited AEs, serious AEs (SAEs), and special interest AEs were collected. Antigen-specific antibody responses were measured using serum enzyme-linked immunosorbent assay. T-cell immune responses were measured using enzyme-linked immunospot and intracellular cytokine staining. No SAEs, deaths, or AEs leading to treatment discontinuation were found. The solicited local and systemic AEs observed were consistent with those previously reported. Compared with adults administered with the placebo, those administered with three intramuscular vaccine injections exhibited significantly higher antigen-specific antibody levels and Type 1 T-helper cellular immune responses. The ID93 + GLA-SE vaccine induced antigen-specific cellular and humoral immune responses, with an acceptable safety profile in previously healthy, BCG-vaccinated, Mycobacterium tuberculosis-uninfected adult healthcare workers. This clinical trial was retrospectively registered on 16 January 2019 at Clinicaltrials.gov (NCT03806686).


A case study in converting disposable process scouting devices into disposable bioreactors as a future bioprocessing tool.
In this study, we perform mass transfer characterization (k(L) a) on a novel mechanically driven/stirred Process Scouting Device, PSD, (SuperSpinner D 1000®, SSD) and demonstrate that this novel device can be viewed as disposable bioreactor. Using patch-based optical sensors, we were able to monitor critical cell culture environmental conditions such as dissolved oxygen (DO) and pH in SSD for comparison to a 1 L standard spinner (SS) flask. We also coupled these mass transfer studies with mixing time studies where we observed relative high mixing times (5.2 min) that are typically observed in production scale bioreactors. Decreasing the mixing time 3.5-fold resulted in 30% increase in k(L) a (from 2.3 to 3.0 h(-1) ) and minimum DO level increased from 0% to 20% for our model hybridoma cell line. Finally, maximum viable cell density and protein titer stayed within ±20% of historical data, from our standard 5 L stirred bioreactor (Biostat®) operated under active DO control.


Production of monoclonal antibodies against phloem-limited prokaryotes of plants: a general procedure using extracts from infected periwinkles as immunogen.
Using Spiroplasma citri-infected periwinkle extracts as the source of antigens, together with monoclonal antibody technology, we determined the conditions for mouse immunization and developed a screening assay to detect those hybridomas which produce immunoglobulins specifically directed against the pathogen.


Markedly elevated hepatic arterial velocity-HAV greater than 200 cm/s-is not specific to hepatobiliary disease.
The aim of the study was to review the different etiologies and outcomes of patients with hepatic artery velocities greater than 200 cm/s. This retrospective study included 88 hospitalized patients in whom angle-corrected proper hepatic artery flow velocities greater than 200 cm/s were obtained during an abdominal ultrasonographic examination. Peak systolic hepatic artery flow velocities, hepatic artery resistance index, and portal vein flow velocities were evaluated. The patients were then allocated to one of four groups based on their primary underlying diagnosis: structural liver disease, nonstructural liver disease, generalized infection, or miscellaneous. The median hepatic artery velocity was similar for all groups, ranging from 226 to 238 cm/s. The maximum portal venous velocities were not significantly different between groups. No lab values were statistically different between the groups, except total bilirubin that was greater in the nonstructural liver disease group (8 mg/dL). Overall, 9/88 (10.2%) of patients with elevated hepatic artery velocity died within 30 days of their ultrasonographic examination. Elevated hepatic artery velocity greater than 200 cm/s in hospitalized patients is not specific to primary hepatobiliary disease but may indicate acute hepatic dysfunction from other causes such as infection or sepsis.


Prescribed opioid use is associated with adverse cardiovascular outcomes in community-dwelling older persons.
Prescribed opioids are commonly used in the older community-dwelling population for the treatment of chronic pain. Although the harmful effects of opioid abuse and overdose are well understood, little is known about the long-term cardiovascular (CV) effects of prescribed opioids. The aim of this study was to investigate the CV effects associated with prescribed opioid use. A post hoc analysis of participants in the Aspirin in Reducing Events in the Elderly (ASPREE) trial was conducted. Participants in the ASPREE trial included community-dwelling older adults without a prior history of CV disease (CVD). Prescribed opioid use was defined as opioid use at baseline and/or at the first annual visit (AV1). Cox proportional hazards regression was used to calculate hazard ratios and 95% confidence intervals (95% CI) for associations between opioid use and CVD events following AV1. Of the 17 701 participants included (mean age 75.2 years, 58.2% female), 813 took opioids either at baseline or at AV1. Over a median follow-up period of 3.58 years (IQR 2.50-4.62), CVD events, most notably heart failure hospitalization, occurred in 7% (n = 57) amongst opioid users and 4% (n = 680) amongst non-opioid users. After adjustment for multiple covariates, opiate use was associated with a 1.67-fold (CI 1.26-2.23, P < 0.001) increase in the hazard ratio for CVD events. These findings identify opioid use as a non-traditional risk factor for CVD events in community-dwelling older adults.


Asymptomatic focal calcium pyrophosphate crystal deposition within partially failed repair tissue after matrix-assisted autologous chondrocyte implantation.
Possible failures of autologous chondrocyte implantation (ACI), a cell-based technique for articular cartilage repair, are not always clinically apparent and the underlying mechanisms largely remain unknown. This case report presents the first scenario in the literature highlighting an association of a medium-term partial failure of an advanced ACI procedure (matrix-assisted ACI) in the knee with focal asymptomatic calcium pyrophosphate deposition disease, a common inflammatory pyrophosphate arthropathy. The specific presence of CPPDs, resulting from increased biomechanical stresses in the repair tissue-cartilage and repair tissue-subchondral bone integration sites, together with the absence of cartilage regeneration was identified and possibly contributed to the partial failure.Level of evidence V.


Monosomy 8 rescue gave cells with a normal karyotype in a mildly affected man with 46,XY,r(8) mosaicism.
The psychomotor and somatic development from early childhood into adult life is described in a man with 46,XY,r(8)/46,XY mosaicism. The ring chromosome 8 appeared to be of normal length on G-banding, but terminal deletions on 8q and 8p were detected with FISH and CGH. By STR marker analysis the 8p deletion proved to be quite large, at least 6.74 Mb, while the 8q deletion was small, around 2.5 Mb. The haplotype analysis also demonstrated that the r(8) originated from a maternal chromosome 8, and that cells with normal male karyotype resulted from monosomy 8 rescue after loss of the ring 8, i.e. a mitotic duplication of the paternal chromosome 8. The patient has a mild phenotype with no malformations and mild mental retardation, also compared to other ring chromosome 8 patients. His clinical condition has remained stable for the last 20 years.


Stimulatory effect of prostaglandins on the production of hexosamine-containing substances by cultured fibroblasts (4) adenosine 3':5'-cyclic monophosphate independent stimulation by prostaglandin F2alpha.
The mechanism of the stimulatory effect of prostaglandin PG) F2alpha on the production of hexosamine-containing substances by cultured fibroblasts was studied with special reference to adenosine 3':5'-cyclic monophosphate (cAMP). At the stationary phase, the cells were exposed for 6 hrs to PGF2alpha, E1, cAMP or dibutyryl-cAMP in a wide range of concentrations. cAMP itself showed a slight stimulation on the production of hexosamine-containing substances, and the effect was enhanced by using the dibutyryl derivative. PGF2alpha had much a greater capacity than either the exogeneous cAMP or the dibutyryl-cAMP for enhancing the production of hexosamine-containing substances. To know whether cAMP is involved in the stimulatory effect of PGF2alpha, intracellular cAMP level was concomitantly measured in both PGF2alpha and PGE1 treated cultures. Although the cellular cAMP level in PGE1 treated cultures was much higher than that in the PGF2alpha treated cultures, the stimulatory effect on the production of hexosamine-containing substances in PGE1 treated cultures was always much smaller than that in the PGF2alpha treated cultures. Moreover, PGF2alpha had a significant stimulatory effect on the production of hexosamine-containing substances even at a low concentration as 100 pg/ml, which is small enough not to increase any cellular cAMP level. From these results, it was concluded that the stimulatory effect of PGF2alpha on the production of hexosamine-containing substances by cultured fibroblasts is not mediated by cAMP and is caused by a mechanism different from that caused by cAMP.


CRYAB predicts clinical prognosis and is associated with immunocyte infiltration in colorectal cancer.
αB-Crystallin (CRYAB) is differentially expressed in various tumors. However, the correlation between CRYAB and immune cell infiltration in colorectal cancer (CRC) remains unclear. Kaplan-Meier survival curves in The Cancer Genome Atlas (TCGA) were used to evaluate the relationship between CRYAB expression and both overall survival and progression-free survival. The relationships between CRYAB expression and infiltrating immune cells and their corresponding gene marker sets were examined using the TIMER database. The expression of CRYAB was lower in CRC tumor tissues than in normal tissues (P < 0.05). High CRYAB gene expression and high levels of CRYAB gene methylation were correlated with high-grade malignant tumors and more advanced tumor, nodes and metastasis (TNM) cancer stages. In addition, in colorectal cancer, there was a positive correlation between CRYAB expression and immune infiltrating cells including neutrophils, macrophages, CD8 + T cells, and CD4 + T cells, as well as immune-related genes including CD2, CD3D, and CD3E. Methylation sites such as cg13084335, cg15545878, cg13210534, and cg15318568 were positively correlated with low expression of CRYAB. Because CRYAB likely plays an important role in immune cell infiltration, it may be a potential tumor-suppressor gene in CRC and a potential novel therapeutic target and predictive biomarker for colorectal cancer (CRC).


Pediatric hospitalizations for upper urinary tract calculi: Epidemiological and treatment trends in the United States, 2001-2014.
The incidence of urolithiasis in the pediatric population in the United States has steadily been increasing over the past few decades. Few studies to date have examined trends in the hospitalization and inpatient surgical treatment of urolithiasis in the pediatric population using nationally representative data. The aim was to evaluate nationwide trends in the rates of pediatric hospitalization and inpatient surgical activity for upper urinary tract calculi (UUTC) in the United States from 2001 to 2014. The National Inpatient Sample (NIS) databases for 2001-2014 were queried. Hospitalizations for patients younger than age 18 (excluding newborns), with principal discharge diagnoses of kidney or ureteral calculi were selected. Surgical procedures during hospitalization were identified. Hospitalization and surgical activity data were analyzed using trends tests, chi-square statistics, and multivariable logistic regression as appropriate. Of an estimated 30.2 million pediatric hospitalizations during the study period, 44,369 overall (147 per 100,000) were for UUTC. The total number and proportion of UUTC hospitalizations per 100,000 all-cause admissions significantly decreased between 2001 and 2014 (p < 0.0001) (figure). Surgical intervention was undertaken in 19,946 (45%) of UUTC hospitalizations, with significantly increasing frequency over the study interval (p < 0.0001). Urinary tract drainage was the most frequently performed surgical intervention. On multivariable analysis, significant predictors of a higher likelihood of undergoing inpatient surgical intervention during hospitalization for UUTC included older age, female gender, deficiency anemias, hypertension, neurologic disorders, paralysis, and hospitalization after 2001. The declining trend in hospitalization for UUTC likely reflects a shift toward outpatient care for routine cases, reserving hospitalization for sicker patients or those with complications of urolithiasis. Similar to previous studies, we also observed that girls were significantly more likely than boys to be hospitalized for stone disease, and that majority of the stone activity in the pediatric population was in children aged 15-17 years. We also observed a sharp increase in the proportion of hospitalized patients who underwent surgical intervention between 2001 and 2014, but the primary driver of this trend remains uncertain. Pediatric hospitalizations for UUTC in US children significantly decreased between 2001 and 2014, while of those hospitalized the proportion who underwent stone-related surgical intervention significantly increased over the same period. A shift towards outpatient care, reserving hospitalization and inpatient surgical care for sicker patients, those with urolithiasis-related complications, or those who fail conservative management, is a possible explanation for these observed trends.


Cholinergic hyperinnervation in the cerebral cortex of microencephalic rats does not result in muscarinic receptor down-regulation or in alteration of receptor-stimulated phosphoinositide metabolism.
Administration of methylazoxymethanol (MAM; 25 mg/kg) to pregnant rats at gestational day 15 (GD 15) induces a marked reduction of telencephalic areas of the offspring brain. Previous neurochemical studies demonstrated a marked cholinergic hyperinnervation in the cerebral cortex of microencephalic rats. In this study we have evaluated whether this cholinergic hyperinnervation could result in altered functionality of muscarinic receptors. Acetylcholinesterase activity (AChE) was increased by 69% in the cerebral cortex of MAM treated rats, confirming a relative hyperinnervation, whereas in the hippocampus and striatum no significant changes were observed. Despite the marked hyperinnervation, in the cerebral cortex of microencephalic rats neither muscarinic receptor-stimulated phosphoinositide metabolism nor muscarinic receptor density were altered. No differences in receptor density were also observed in the hippocampus and striatum. Chronic diisopropylfluorophosphate (DFP) administration induced a marked decrease of AChE activity and down-regulation of muscarinic receptors whereas atropine administration resulted in receptor up-regulation in cerebral cortex, striatum and hippocampus of both control and MAM rats. The results confirm a relative cholinergic hyperinnervation in the cerebral cortex of microencephalic rats and demonstrate that the regulation of muscarinic receptor-stimulated phosphoinositide metabolism and muscarinic receptor plasticity is not modified in a condition of increased cholinergic presynaptic terminals.


Biotoxicity of mercury to Chlorella vulgaris as influenced by amino acids.
The toxicity of mercury ion, on Chlorella vulgaris, is largely influenced by amino acids. Five amino acids, namely alanine, asparagine, glutamic acid, cysteine and histidine, were added separately to the medium containing static dose of mercury. Survival (%) of the alga was reduced with the increasing concentrations of mercury. Of these five amino acids, cysteine was found to be the most effective while alanine and glutamic acid were the least effective on reducing the toxic effect of mercury on the alga measured in terms of growth, chlorophyll and protein content. The order of detoxification was Alanine < Glutamate < Asparargine < Histidine < Cysteine. Amino acids from ligands with Hg2+ making it less toxic to the alga and produce an additional source of energy for growth and development.


Electromyographic study of human jaw-closing muscle endurance, fatigue and recovery at various isometric force levels.
The effect of a sustained isometric clenching, at various force levels or maximum voluntary bite-force levels, was evaluated on normal jaw-function subjects. Surface electromyographic (EMG) recordings of the masseter and temporalis were taken and bite force was measured using an intra-oral force transducer placed unilaterally in the 2nd premolar-1st molar region. Subjects sustained isometric force at the 25, 50, 75 and 100 per cent force level. Maximal bite force and EMG contraction levels were measured before, during and after these sustained isometric tasks. Subjects showed no change in their brief maximal contraction or force levels during or after various fatigue-inducing isometric tasks. The findings suggested a lack of contractile failure in the jaw-closing muscles. Pain intolerance rather than neuromuscular fatigue is the limiting factor of a sustained submaximal or even maximal clenching effort.


Detection of space-time clusters and epidemiological examinations of scabies in chamois.
In this paper some methodological approaches towards the detection of temporal, spatial and space-time clusters are presented and discussed in context with epidemiological factors of scabies in chamois. This presentation comprises in particular the use of the Scan Test (Wallenstein, 1980) for the detection of temporal clusters and the tests developed by Knox (1964) and Mantel (1967) for the testing of spatiotemporal interactions. Between 1952 and 1998 a total of 1689 cases of scabies in chamois and six cases in ibex were documented in 137 game preserves of the Austrian province of Styria. The evaluations indicate the presence of spatial and temporal clusters. Some possibilities for the spread of scabies in chamois are presented and reveal ibex as a probable vector for scabies transmission between chamois populations. The information may lead to strategies for controlling the spread of the disease.


Effect of organophosphorus hydrolysing enzymes on obidoxime-induced reactivation of organophosphate-inhibited human acetylcholinesterase.
The reactivation of organophosphate (OP)-inhibited acetylcholinesterase (AChE) by oximes results inevitably in the formation of highly reactive phosphyloximes (POX), which may re-inhibit the enzyme. An impairment of net reactivation by stable POX was found with 4-pyridinium aldoximes, e.g. obidoxime, and a variety of OP compounds. In this study the effect of organophosphorus hydrolase (OPH), organophosphorus acid anhydrolase (OPAA) and diisopropylfluorophosphatase (DFPase) on obidoxime-induced reactivation of human acetylcholinesterase (AChE) inhibited by different OPs was investigated. Reactivation of paraoxon-, sarin-, soman- and VX-inhibited AChE by obidoxime was impaired by POX-induced re-inhibition whereas no deviation of pseudo first-order kinetics was observed with tabun, cyclosarin and VR. OPH prevented (paraoxon) or markedly reduced the POX-induced re-inhibition (VX, sarin, soman), whereas OPAA and DFPase were without effect. Additional experiments with sarin-inhibited AChE indicate that the POX hydrolysis by OPH was concentration-dependent. The activity of OP-inhibited AChE was not affected by OPH in the absence of obidoxime. In conclusion, OPH may be a valuable contribution to the therapeutic regimen against OP poisoning by accelerating the degradation of both the parent compound, OP, and the reaction product, POX.


Mapping the neuroanatomical impact of very preterm birth across childhood.
Those born very preterm (VPT; <32 weeks gestational age) have an increased risk in developing a wide range of cognitive deficits. In early-to-late childhood, brain structure has been shown to be altered in VPT compared to full-term (FT) children; however, the results are inconsistent. The current study examined subcortical volumes, cortical thickness, and surface area in a large cohort of VPT and FT children aged 4-12 years. Structural magnetic resonance imaging (MRI) was obtained on 120 VPT and 146 FT children who returned up to three times, resulting in 176 VPT and 173 FT unique data points. For each participant, Corticometric Iterative Vertex-based Estimation of Thickness was used to obtain global measurements of total brain, cortical grey and cortical white matter volumes, along with surface-based measurements of cortical thickness and surface area, and Multiple Automatically Generated Templates (MAGeT) brain segmentation tool was used to segment the subcortical structures. To examine group differences and group-age interactions, mixed-effects models were used (controlling for whole-brain volume). We found few differences between the two groups in subcortical volumes. The VPT children showed increased cortical thickness in frontal, occipital and fusiform gyri and inferior pre-post-central areas, while thinning occurred in the midcingulate. Cortical thickness in occipital regions showed more rapid decreases with age in the VPT compared to the FT children. VPT children also showed both regional increases, particularly in the temporal lobe, and decreases in surface area. Our results indicate a delayed maturational trajectory in those born VPT.


The role of band 4.1 in the association of actin with erythrocyte membranes.
Spectrin stimulates the association of F-actin with erythrocyte inside-out vesicles. Although inside-out vesicles are nearly devoid of two of the three major cytoskeletal proteins, spectrin and actin, they retain nearly all of the cytoskeletal protein designated band 4.1. Inside-out vesicles which have been substantially depleted of band 4.1 by extraction in 1 M KCl, 0.4 M urea and then reconstituted with spectrin show a markedly diminished ability to bind actin by comparison with vesicles containing normal amounts of band 4.1. This diminution is not due to an impaired ability of the vesicles to bind spectrin. Addition of purified band 4.1 to vesicles either before or after they have been reconstituted with spectrin restores their actin binding capacity to near normal levels as does addition of a spectrin-band 4.1 complex prepared by sucrose gradient centrifugation. Band 4.1 bound to vesicles in the absence of added spectrin has no effect on actin binding. Our results suggest that a spectrin band 4.1 complex is responsible for binding actin to erythrocyte membranes.


The development of genome-wide single nucleotide polymorphisms in blue wildebeest using the DArTseq platform.
Blue wildebeest (Connochaetes taurinus taurinus) are economically important antelope that are widely utilised in the South African wildlife industry. However, very few genomic resources are available for blue wildebeest that can assist in breeding management and facilitate research. This study aimed to develop a set of genome-wide single nucleotide polymorphism (SNP) markers for blue wildebeest. The DArTseq genotyping platform, commonly used in polyploid plant species, was selected for SNP discovery. A limited number of published articles have described the use of the DArTseq platform in animals and, therefore, this study also provided a unique opportunity to assess the performance of the DArTseq platform in an animal species. A total of 20,563 SNPs, each located within a 69 bp sequence, were generated. The developed SNP markers had a high average scoring reproducibility (>99%) and a low percentage missing data (~9.21%) compared to other reduced representation sequencing approaches that have been used in animal studies. Furthermore, the number of candidate SNPs per nucleotide position decreased towards the 3' end of sequence reads, and the ratio of transitions (Ts) to transversions (Tv) remained similar for each read position. These observations indicate that there was no read position bias, such as the identification of false SNPs due to low sequencing quality, towards the tail-end of sequencing reads. The DArTseq platform was also successful in identifying a large number of informative SNPs with desirable polymorphism parameters such as a high minor allele frequency (MAF). The Bos taurus genome was used for the in silico mapping of the marker sequences and a total of 6020 (29.28%) sequences were successfully mapped against the bovine genome. The marker sequences mapped to all of the bovine chromosomes establishing the genome-wide distribution of the SNPs. Moreover, the high observed Ts:Tv ratio (2.84:1) indicate that the DArTseq platform targeted gene-rich regions of the blue wildebeest genome. Finally, functional annotation of the marker sequences revealed a wide range of different putative functions indicating that these SNP markers can be useful in functional gene studies. The DArTseq platform, therefore, represents a high-throughput, robust and cost-effective genotyping platform, which may find adoption in several other African antelope and animal species.


Classical conditioning leads to changes in extracellular concentrations of ependymin in goldfish brain.
ELISA measurements showed that brain extracellular fluid (ECF) levels of ependymin decreased for animals that learned to associate a paired presentation of a light stimulus (CS) with the onset of an electric shock (US), whereas no changes were obtained for control goldfish that received the same number of stimuli delivered in a random unpaired order. Studies of the time course of the changes showed an immediate decrease (19%) after training followed by an increase (20%) above baseline by 5 h and a final return to baseline by 25 h. These data extend the findings of previous experiments, which demonstrated a role for ependymin in two training procedures that involved motor learning, to classical conditioning where no motor learning occurs. Thus it appears that ependymin may have a functional role in molecular mechanisms of learning and memory in general.


Episiotomy characteristics and risks for obstetric anal sphincter injuries: a case-control study.
To investigate the association between the geometrical properties of episiotomy and obstetric anal sphincter injuries (OASIS) because episiotomies angled at 40-60° are associated with fewer OASIS than episiotomies with more acute angles. Case-control study. University Hospital of North Norway, Tromsø and Nordland Hospital, Bodø, Norway. Seventy-four women who had one vaginal birth and episiotomy. Cases (n = 37) have sustained OASIS at birth, while controls (n = 37) had not. The groups were matched for instrumental delivery. Two groups of women with history of only one vaginal birth were compared. Episiotomy scar was identified and photographed and relevant measures were taken. Data were analysed using conditional logistic analysis. Mean episiotomy angle, length, depth, incision point. The risk of sustaining OASIS decreased by 70% (odds ratio [OR] 0.30; 95% CI 0.14-0.66) for each 5.5-mm increase in episiotomy depth, decreased by 56% (OR 0.44; 95% CI 0.23-0.86) for each 4.5-mm increase in the distance from the midline to the incision point of the episiotomy, and decreased by 75% (OR 0.25; 95% CI 0.10-0.61) for each 5.5-mm increase in episiotomy length. Lastly, there was no difference in mean angle between groups but there was a "U-shaped" association between angle and OASIS (OR 2.09; 95% CI 1.02-4.28) with an increased risk (OR 9.00; 95% CI 1.1-71.0) of OASIS when the angle was either smaller than 15° or >60°. The present study showed that scarred episiotomies with depth > 16 mm, length > 17 mm, incision point > 9 mm lateral of midpoint and angle range 30-60° are significantly associated with less risk of OASIS. Shrinkage of tissue must be considered.


The black death past and present. 2. Some historical problems.
This paper looks, from a historian's point of view, at the black death and the epidemics of plague which succeeded it in Europe from the fourteenth to the seventeenth centuries. It identifies the controversial questions, of medical as well as historical interest, which have been raised by recent work. These include the origins of plague epidemics, the role of rodents and insect vectors in them, and the reasons for their disappearance from western Europe.


Malignant hyperthermia-like reactions in three anesthetized horses.
Problems encountered during halothane anesthesia in 3 horses included increasing rectal temperature, muscle rigidity or movement during anesthesia, irregular breathing patterns, and difficulty in stabilizing blood pressure. One horse had prolonged muscle fasciculations and failure to relax after administration of succinylcholine in addition to halothane. That horse developed severe, bilateral myositis of the triceps, lumbar, and gluteal muscles. Problems encountered were similar to those caused by the disease known as malignant hyperthermia.


Circular RNA circ_0058123 Targets the miR-939-5p/RAC1 Pathway to Promote the Development of Colorectal Cancer.
Circular RNA (circRNA) can be used as a potential target for cancer treatment. However, the biological function and potential molecular mechanism of circ_0058123 in the development of colorectal cancer (CRC) are still unclear. The expression levels of circ_0058123, microRNA-939-5p (miR-939-5p) and Rac family small GTPase 1 (RAC1) were measured by quantitative real-time polymerase chain reaction or western blot assay. 5-Ethynyl-2'-deoxyuridine (EdU) incorporation assay, transwell assay, tube formation assay and flow cytometry apoptosis assay were conducted to assess CRC cell functions. In addition, protein expression was measured with western blot assay. Dual-luciferase reporter assays and RNA immunoprecipitation assay were conducted to confirm the relationships between miR-939-5p and circ_0058123, and miR-939-5p and RAC1. In vivo CRC tumor growth experiment also were carried out to determine circ_0058123-mediatede effects on tumor formation. Our data showed that circ_0058123 and RAC1 expression were increased, but miR-939-5p was decreased in both of CRC tissues and cell lines. Circ_0058123 depletion repressed CRC cell proliferation, migration, invasion and tube formation but promoted cell apoptosis. Down-regulation of circ_0058123 could significantly suppress the CRC progression, while the addition of miR-939-5p inhibitor could reverse this effect. Circ_0058123 directly targeted miR-939-5p, and RAC1 was a target of miR-939-5p. Furthermore, RAC1 overexpression could rescue the effect of miR-939-5p on CRC development. Lastly, silence of circ_0058123 inhibited CRC tumor growth in vivo. In conclusion, circ_0058123 could promote CRC progression through regulating the miR-939-5p/RAC1 axis and may be a valuable biomarker for early diagnosis and prognosis of CRC.


Extending indication: role of living donor liver transplantation for hepatocellular carcinoma.
Living donor liver transplantation. Indication of hepatocellular carcinoma. Milan criteria. United Network for Organ Sharing (UNOS) tumor staging. Alfa-fetoprotein (AFP). Protein induced by vitamin K absence or antagonism factor II (PIVKA II).


Clinico-pathologic predictors of patterns of residual disease following neoadjuvant chemotherapy for breast cancer.
Among breast cancer patients treated with neoadjuvant chemotherapy (NAC) who do not experience a pathologic complete response (pCR), the pattern of residual disease in the breast varies. Pre-treatment clinico-pathologic features that predict the pattern of residual tumor are not well established. To investigate this issue, we performed a detailed review of histologic sections of the post-treatment surgical specimens for 665 patients with stage I-III breast cancer treated with NAC followed by surgery from 2004 to 2014 and for whom slides of the post-NAC surgical specimen were available for review. This included 242 (36.4%) patients with hormone receptor (HR)+/HER2- cancers, 216 (32.5%) with HER2+ tumors, and 207 (31.1%) with triple negative breast cancer (TNBC). Slide review was blinded to pre-treatment clinico-pathologic features. pCR was achieved in 7.9%, 37.0%, and 37.7%, of HR+/HER2- cancers, HER2+ cancers, and TNBC respectively (p < 0.001). Among 389 patients with residual invasive cancer in whom the pattern of residual disease could be assessed, 287 (73.8%) had a scattered pattern and 102 (26.2%) had a circumscribed pattern. In both univariate and multivariate analyses, there was a significant association between tumor subtype and pattern of response. Among patients with HR+/HER2- tumors, 89.4% had a scattered pattern and only 10.6% had a circumscribed pattern. In contrast, among those with TNBC 52.8% had a circumscribed pattern and 47.2% had a scattered pattern (p < 0.001). In addition to subtype, histologic grade and tumor size at presentation were also significantly related to the pattern of residual disease in multivariate analysis, with lower grade and larger size each associated with a scattered response pattern (p = 0.002 and p = 0.01, respectively). A better understanding of the relationship between pre-treatment clinico-pathologic features of the tumor and pattern of residual disease may be of value for helping to guide post-chemotherapy surgical management.


Establishment of substituent effects in the DNA binding subunit of CBI analogues of the duocarmycins and CC-1065.
An extensive series of CBI analogues of the duocarmycins and CC-1065 exploring substituent effects within the first indole DNA binding subunit is detailed. In general, substitution at the indole C5 position led to cytotoxic potency enhancements that can be >/=1000-fold providing simplified analogues containing a single DNA binding subunit that are more potent (IC(50)=2-3 pM) than CBI-TMI, duocarmycin SA, or CC-1065.


Repeated attacks of type III hereditary angioedema with factor XII mutation during pregnancy.
In type III hereditary angioedema (HAE type III), the phenotype is the same as type I and type II disease, but the level and function of C1-esterase inhibitor (C1-INH) is normal. Hereditary angioedema type III has been described as an oestrogen-sensitive form because it can be triggered or aggravated by exposure to high oestrogen levels as seen during pregnancy, especially when associated with Factor XII mutation. This case report describes the evolution and management of repeated angioedema attacks during pregnancy in a woman with HAE, with normal levels and function of C1-INH (type III); and a mis-sense mutation of factor XII. The physiopathology and genetic features, the unpredictability of clinical manifestations and the management during pregnancy and delivery are discussed.


In vitro enzymatic degradation of a biological tissue fixed by alginate dialdehyde.
Biological tissues must be chemically fixed before they can be implanted in humans as tissue engineering scaffolds. To provide an ideal tissue engineering scaffold material, which is biodegradable and cytocompatible, a novel crosslinking agent, alginate dialdehyde (ADA), was employed to fix biological tissues by our group. The study mainly investigated the enzymatic degradation of ADA fixed biological tissues in vitro. Glutaraldehyde, the most commonly used crosslinking agent for biological tissue fixation, was employed as a control. The results suggested that, the ADA fixation could enhance the resistance against enzymatic degradation of biological tissues effectively. Meanwhile, compared to glutaraldehyde-fixed tissues, the ADA-fixed tissues could also degrade gradually over time. Moreover, the ADA crosslinking reagent itself had a stimulatory effect on cell proliferation when at an appropriate concentration. The results obtained in this study demonstrate that ADA fixation might provide a successful example of the biodegradable scaffold materials in tissue engineering.


Effect of L-cysteine on the oxidation of cyclohexane catalyzed by manganeseporphyrin.
Effect of L-cysteine as the cocatalyst on the oxidation of cyclohexane by tert-butylhydroperoxide (TBHP) catalyzed by manganese tetraphenylporphyrin (MnTPP) has been investigated. The results showed that L-cysteine could moderately improve the catalytic activity of MnTPP and significantly increase the selectivity of cyclohexanol. Different from imidazole and pyridine, the L-cysteine may perform dual roles in the catalytic oxidation of cyclohexane. Besides as the axial ligand for MnTPP, the L-cysteine could also react with cyclohexyl peroxide formed as the intermediate to produce alcohol as the main product.


African Swine Fever Virus pF778R Attenuates Type I Interferon Response by Impeding STAT1 Nuclear Translocation.
African swine fever virus (ASFV) is an extensive and intricate double-stranded DNA virus with approximately 100% lethality in domestic swine. There is no effective vaccine to combat this virus, and this has led to substantial economic losses in the swine industry. ASFV encodes various proteins that impede interferon-based immune defenses in the host by employing diverse mechanisms. However, the roles of most of these proteins remain unknown. Therefore, understanding the immune evasion mechanisms employed by ASFV may facilitate the development of effective measures against the virus. In this study, we discovered a negative regulation of the type I interferon (IFN) response by the ASFV ribonuclease reductase large subunit pF778R. This novel type Ⅰ IFN response antagonist significantly inhibits IFN-α-induced interferon-stimulated response element promoter activation, precludes the upregulation of various interferon-stimulated genes, and prevents STAT1 nuclear translocation. Mechanistically, pF778R did not affect the protein levels of crucial molecules in the JAK/STAT signaling pathway or engage in direct interactions. However, pF778R expression impedes type I IFN responses mediated by the JAK/STAT signaling pathway. Further investigations revealed that pF778R did not interfere with STAT1 phosphorylation or dimerization, but it inhibited IFN signaling by weakening the nuclear accumulation of activated STAT1. The critical role of the ASFV protein pF778R in evading IFN-I-mediated innate immunity highlights a unique mode of ASFV evasion and provides insights into the pathogenic mechanism of the virus.


No effect of genetic obesity and mazindol on imidazoline I2 binding sites in the brain of Zucker rats.
The density and affinity states of imidazoline I2 binding sites as well as the density of alpha 2-adrenoceptors were quantitated in the brain of lean and obese Zucker rats. No significant differences were obtained between Zucker phenotypes for these receptors in the cerebral cortex and hypothalamus. Moreover, chronic treatment with the anorexic imidazoline derivative, mazindol, did not alter the density of brain imidazoline I2 binding sites. It is concluded that this genetic model of obesity is not associated with abnormal imidazoline I2 binding sites.


Edge detection at multiple locations using a 'radar' tracking algorithm as exemplified in isolated guinea-pig lymphatic vessels.
An algorithm based on the Kalman filter and used for tracking a target in a noisy environment has been applied to monitoring the position of moving boundaries as recorded by a video camera. The method, termed the Probabilistic Data Association Filter, is computationally efficient, allowing simultaneous monitoring at many positions along the boundary at video frame rates of 25 Hz. The present study uses the tracking procedure to investigate and correlate constrictions along lymphatic vessels. The procedure is well suited for real time tracking of visualised biological events including the simultaneous monitoring of constrictions in networks of blood vessels, the movement of cells or the trajectory of intracellular events monitored by intracellular fluorescent markers.


Genes and pathways differentially expressed in the brains of Fxr2 knockout mice.
Fragile X syndrome is a common inherited form of mental retardation and originates from the absence of expression of the FMR1 gene. This gene and its two homologues, FXR1 and FXR2, encode for a family of fragile X related (FXR) proteins with similar tissue distribution, together with sequence and functional homology. Based on these characteristics, it has been suggested that these proteins might partly complement one another. To unravel the function of Fxr2 protein, the expression pattern of 12,588 genes was studied in the brains of wild-type and Fxr2 knockout mice, an animal model which shows behavioral abnormalities partly similar to those observed in Fmr1-knockout mice. By genome expression profiling and stringent significance tests we identify genes and gene groups de-regulated in the brains of Fxr2 knockout mice. Differential expression of candidate genes was validated by real-time PCR, in situ hybridization, immunohistochemistry and western blot analysis. A number of differentially expressed genes associated with the Fxr2 phenotype have been previously involved in other memory or cognitive disorders.


[Diagnosis and treatment of esophageal leiomyoma].
To summarize the experience of diagnosis and treatment of esophageal leiomyoma. Clinical data of 52 patients with esophageal leiomyoma were analyzed from 1993 to 2002. About 54% patients in this group had difficulty of food intake. The diagnostic accuracy of gastrointestinal barium meal series, computed tomography, gastric endoscope and endoscopic ultrasonography (EUS) for esophageal leiomyoma was 64% 44% 27% and 90% respectively. All patients received operation, resection of esophageal leiomyoma by videothoracoscopy (VAS) and endoscope were performed in 6, 9 patients respectively. The remaining 37 patients received regular open operation,in whom 32 cases received enucleation of esophageal leiomyoma, 5 cases received partial esophageal resection and esophageal-gastric anastomosis. No serious complications occurred except only one case needed operation again because of bleeding. EUS is an effective method for diagnosing esophageal leiomyoma. VAS and endoscopic treatment should be considered for suitable cases in order to reduce the trauma.


Clinical and radiographic outcomes of a trabecular titanium™ acetabular component in hip arthroplasty: results at minimum 5 years follow-up.
Aim of this prospective study was to evaluate mid-term clinical and radiographic outcomes in total hip arthroplasty using an acetabular cup made of an innovative biomaterial, Trabecular Titanium™, whose highly porous structure and mechanical properties have been designed to mimic those of the natural bone, thus promoting a more physiological load transfer and a more durable fixation. Between September 2007 and November 2009, 134 total hip replacements and eight revisions were carried out using DELTA-TT primary cups (Lima Corporate, Villanova di San Daniele del Friuli, Italy) in 133 consecutive patients. Mean age was 57.5 ± 14.7 SD (18-92) years. Diagnosis was primarily hip osteoarthritis in 85 (63 %) cases, developmental dysplasia of the hip (DDH) in 24 (18 %) and hip avascular necrosis (AVN) in 10 (7 %). All the revision procedures were due to aseptic loosening of the original implant. Approval of the Institutional Review Board of the IRCCS Policlinico San Matteo in Pavia was obtained for this study. Mean follow-up was 72.7 ± 7.9 SD (60-86) months. Average Harris Hip Score (HHS) significantly increased from 44.2 ± 5.4 SD (35-52) preoperatively to 95.9 ± 3.5 SD (88-100) at the last follow-up. No major post-operative complications were observed. 99.3 % of the acetabular components were radiographically stable at the last follow-up, without any radiolucent lines, sclerotic areas or periprosthetic osteolysis. Kaplan-Meier survival rate was 99.3 % at 5 years (95 % confidence interval). This first account on the mid-term clinical performance of the DELTA-TT cup shows primary and secondary stability, thus representing an optimal solution for patients with high demands or affected by severe hip conditions.


Primary central nervous system lymphoma: current state of anti-CD20 therapy and appraisal of reported response criteria.
Primary central nervous system lymphoma (PCNSL) is an aggressive non-Hodgkin's lymphoma which is confined to the central nervous system and may also affect intraocular structures. Despite high initial rates of response to methotrexate-based chemotherapy, more than 50% of patients will experience relapse and about 10% have disease that is refractory to chemotherapy. Outcome in patients who fail treatment is very poor, and therefore new therapeutic approaches that may increase the rate of complete response and the proportion of durable remission are sought. Based on the pivotal role that anti-CD20 therapy now plays in the treatment outcome of aggressive systemic B-cell lymphomas, a similar approach is commonly being adapted for PCNSL despite the lack of evidence for its effectiveness. This review examines the current status and level of evidence for the use of monoclonal antibodies against the CD20 surface antigen, which is present on normal and malignant B-cells in PCNSL. The review covers both systemic and local (intracerebrospinal fluid or intravitreal) administration of CD20 monoclonal antibodies in PCNSL. In addition, it scrutinizes the response criteria commonly reported for evaluation of treatment outcome. The importance of differentiating unconfirmed complete response from partial response is outlined and the lack of consensus on response criteria for atypical imaging presentations of PCNSL is delineated.


An improved microwave assisted sequential extraction method followed by spectrometric analysis for metal distribution determination in South African coal samples.
Some metal pollutants are corrosive in nature, are associated with fouling and slagging challenges of the coal boilers, are highly volatile and might cause air pollution and are catalyst poisoners during Fischer-Tropsch catalytic reaction. Therefore, this work describes an improved microwave-assisted sequential extraction (MW-ASE) method followed by ICP-OES/MS analysis for metal distribution determination in South African coal samples. The multivariate optimum conditions for each sequential step were 0.1 g, 200 °C and 5 min for sample amount, microwave temperature and extraction time, respectively. Under the optimum conditions, Ga, Sr and Ba were the only metals that showed solubility towards water, therefore, these metals are classified as highly mobile and eco-toxic under wet environmental conditions. Additionally, all the investigated metals showed solubility towards acidic conditions (HCl and HNO3). These results suggest that, most metal ions are predominantly bonded to sulphate, sulphide, and carbonate coal minerals. Alternatively, Ce, Cr and Y showed total extraction recoveries of ≤ 90%, confirming their strong affinity towards quartz coal minerals. In overall, the proposed MW-ASE method reported short extraction time (0.34 h), environmentally friendly reagents (H2O and diluted H2O2) and rapid multivariate optimization with acceptable extraction efficiencies (79-98%) and reproducibility (RSD ≤ 5%).


Circulating Carbonic Anhydrase IX and Antiangiogenic Therapy in Breast Cancer.
Carbonic anhydrase IX (CAIX) is a hypoxia regulated metalloenzyme integral to maintaining cellular pH. Increased CAIX expression is associated with poor prognosis in breast cancer. To explore CAIX as a biomarker for breast cancer therapies, we measured plasma CAIX levels in healthy control subjects and in breast cancer patients. In control subjects we evaluated plasma CAIX stability via commercially available ELISA. We then similarly quantified plasma CAIX levels in (1) locally advanced breast cancer (LABC) patients treated with neoadjuvant paclitaxel + sunitinib (T + S) followed by doxorubicin and cyclophosphamide (AC); (2) metastatic breast cancer (MBC) patients treated with systemic chemotherapy. Plasma CAIX levels were stable at room temperature for at least 48 hours in control subjects. Mean baseline plasma CAIX levels were lower in controls compared to patients with LABC or MBC. In LABC, CAIX levels rose significantly in response to administration of antiangiogenic therapy (T + S) (p = 0.02) but not AC (p = 0.37). In patients with MBC treated without an antiangiogenic agent CAIX levels did not change with therapy. Our results suggest that CAIX may be an easily obtained, stable measure of tumor associated hypoxia as well as a useful pharmacodynamic biomarker for antiangiogenic therapy.


Modeling insights into SARS-CoV-2 respiratory tract infections prior to immune protection.
Mechanistic insights into human respiratory tract (RT) infections from SARS-CoV-2 can inform public awareness as well as guide medical prevention and treatment for COVID-19 disease. Yet the complexity of the RT and the inability to access diverse regions pose fundamental roadblocks to evaluation of potential mechanisms for the onset and progression of infection (and transmission). We present a model that incorporates detailed RT anatomy and physiology, including airway geometry, physical dimensions, thicknesses of airway surface liquids (ASLs), and mucus layer transport by cilia. The model further incorporates SARS-CoV-2 diffusivity in ASLs and best-known data for epithelial cell infection probabilities, and, once infected, duration of eclipse and replication phases, and replication rate of infectious virions. We apply this baseline model in the absence of immune protection to explore immediate, short-term outcomes from novel SARS-CoV-2 depositions onto the air-ASL interface. For each RT location, we compute probability to clear versus infect; per infected cell, we compute dynamics of viral load and cell infection. Results reveal that nasal infections are highly likely within 1-2 days from minimal exposure, and alveolar pneumonia occurs only if infectious virions are deposited directly into alveolar ducts and sacs, not via retrograde propagation to the deep lung. Furthermore, to infect just 1% of the 140 m2 of alveolar surface area within 1 week, either 103 boluses each with 106 infectious virions or 106 aerosols with one infectious virion, all physically separated, must be directly deposited. These results strongly suggest that COVID-19 disease occurs in stages: a nasal/upper RT infection, followed by self-transmission of infection to the deep lung. Two mechanisms of self-transmission are persistent aspiration of infected nasal boluses that drain to the deep lung and repeated rupture of nasal aerosols from infected mucosal membranes by speaking, singing, or cheering that are partially inhaled, exhaled, and re-inhaled, to the deep lung.


Inhibitory effect of triamcinolone acetonide on corneal neovascularization.
Corneal neovascularization (NV) plays an important role in the pathogenesis of corneal disorders. Recently, triamcinolone acetonide (TA) has been reported as a potential treatment for ocular angiogenesis. However, there are no reports on the inhibitory effect of TA on the corneal NV. Triamcinolone acetonide (2 mg) was administered to four rabbits' eyes by a subconjunctival injection immediately after a basic fibroblast growth factor (bFGF)-pellet was placed into the cornea. As a control, four eyes received an injection of distilled water. Four weeks later, the inhibition of corneal NV was evaluated as the percentage ratio of the vessel invasion area to the area that was sandwiched between the pellet and the limbus cornea. To identify the characteristic appearance of new corneal vessels, the control cornea was examined by using the antibody of vascular endothelial growth factor (VEGF). To confirm TA concentration in TA-treated corneas, the TA level was measured using high-performance liquid chromatography. Neovascularization from the limbus to the pellet was detected in control eyes 4 weeks after the bFGF pellet implantation. TA-treated eyes demonstrated the inhibition of the neovascular response to the pellet. The severity of NV as compared between control and TA-treated eyes was statistically significant (P<0.05). Morphologically, new vessel growth was shown in the control cornea, and endothelial cells of new vessels were positively stained with the antibody of VEGF. TA concentration in TA-treated corneas at 2 weeks showed 63.5+/-42.8 microg/g (n=4, mean +/- SD), while TA was not detected in control and TA-treated corneas at 4 weeks. The level of TA was effectively maintained for at least 2 weeks after the subconjunctival injection. We have demonstrated that subconjunctival TA administration inhibited rabbit corneal NV. This agent may prove useful in the treatment of corneal angiogenic disorders.


[Glucose-lowering therapy in patients with cardiac comorbidities].
The risk for cardiovascular events, congestive heart failure and cardiac arrhythmia is significantly increased in patients with diabetes. Although poor glycaemic control has been associated with an increased cardiovascular event rate, aggressive glucose-lowering strategies have failed to improve cardiovascular endpoints or mortality. Therefore, treatment-associated adverse effects, especially hypoglycaemia and weight gain, must be carefully outbalanced against the potential benefits of better glycaemic control. Furthermore, certain drug-specific aspects must be considered: Pioglitazone is contraindicated in patients with heart failure, and DPP-4 inhibitors have recently been associated with an increased heart failure rate. Heart rate may increase during treatment with GLP-1 analogues. Only with metformin a reduction in cardiovascular endpoint has been demonstrated in patients with diabetes. Insulin and sulphonylureas have yielded neutral results in the available endpoint trials. Endpoint studies with GLP-1 analogues or SGLT-2 inhibitors have not yet been completed. These various drug-specific actions in the cardiovascular system need to be born in mind for the choice of the optimal glucose-lowering strategy in patients with cardiac comorbidities.


Targeting 6-phosphogluconate dehydrogenase in the oxidative PPP sensitizes leukemia cells to antimalarial agent dihydroartemisinin.
The oxidative pentose phosphate pathway (PPP) is crucial for cancer cell metabolism and tumor growth. We recently reported that targeting a key oxidative PPP enzyme, 6-phosphogluconate dehydrogenase (6PGD), using our novel small-molecule 6PGD inhibitors Physcion and its derivative S3, shows anticancer effects. Notably, humans with genetic deficiency of either 6PGD or another oxidative PPP enzyme, glucose-6-phosphate dehydrogenase, exhibit non-immune hemolytic anemia upon exposure to aspirin and various antimalarial drugs. Inspired by these clinical observations, we examined the anticancer potential of combined treatment with 6PGD inhibitors and antimalarial drugs. We found that stable knockdown of 6PGD sensitizes leukemia cells to antimalarial agent dihydroartemisinin (DHA). Combined treatment with DHA and Physcion activates AMP-activated protein kinase, leading to synergistic inhibition of human leukemia cell viability. Moreover, our combined therapy synergistically attenuates tumor growth in xenograft nude mice injected with human K562 leukemia cells and cell viability of primary leukemia cells from human patients, but shows minimal toxicity to normal hematopoietic cells in mice as well as red blood cells and mononucleocytes from healthy human donors. Our findings reveal the potential for combined therapy using optimized doses of Physcion and DHA as a novel antileukemia treatment without inducing hemolysis.


17beta-estradiol suppresses cytotoxicity and proliferative capacity of murine splenic NK1.1+ cells.
In order to clarify the effects of 17beta-estradiol (E2) on natural killer (NK) cells and the possibly regulatory mechanisms, we obtained highly purified and viable NK cells from C57BL/6J mouse spleen by a magnetic cell sorter (MACS). These cells were treated with E2 and then their cytotoxicity and proliferative capacity were examined. To further investigate the mechanisms on the effect of E2 on NK cells, expressions of activation-associated markers (CD69, CD122) and inhibitory receptors (CD94, Ly49), and intracellular cytokine production were analyzed. At last, we performed the cDNA microarray to explore the possible involved genes. We found that E2 could suppress NK cell cytotoxicity and proliferative capacity in vitro. E2 reduced NK cell cytotoxicity and proliferative capacity, which may be through influencing the phenotypes and cytokine expression of NK cells, mainly involving CD94 and IFN-gamma. Furthermore, regulation of Stat4, Fyn, Sh2d1a, Eat2, Cd244, Irf1, Runx1, Irf7, Irf5, Esrra and Nr5a1 genes may be related to the cytotoxicity, proliferation and cytokine production of E2-mediated purified NK cells.


Aflatoxins in sunflower seeds: effect of zinc in aflatoxin production by two strains of Aspergillus parasiticus.
Growth and aflatoxin production by Aspergillus parasiticus NRRL 2999 and Aspergillus parasiticus RC 12 were studied both in sunflower seed and a synthetic culture medium (with and without zinc enrichment). On a synthetic culture medium the strains behaved in different ways according to the zinc concentration. In sunflower seed medium the influence of zinc was not so evident. Thus the results show that the influence of zinc is not the same for different strains and substrates.


Effects of the Personalized Recovery Oriented Services (PROS) Program on Hospitalizations.
Though the recovery model has been implemented widely in outpatient mental health settings, there are no large sample evaluations of recovery oriented psychiatric rehabilitation programs that address both serious mental illness (SMI) and co-occurring disorders (COD) using a more comprehensive Medicaid reimbursable approach. This study examined preliminary hospitalization outcomes, for adults with SMI and COD enrolled in the NYS Personalized Recovery Oriented Services (PROS) program. McNemar's chi-square test was used to examine changes in hospitalization rates from pre-PROS admission to post-PROS discharge in a sample of 12,006 adults discharged from PROS. Negative binomial regression models were used to calculate adjusted rates of hospitalizations and hospital days. Demographic, psychosocial, and diagnosis predictor variables were extracted from the OMH web-based Child and Adult Integrated Reporting System. Hospitalization data were extracted from the Mental Health Automated Recordkeeping System, and Medicaid. From pre-admission to post-discharge, psychiatric hospitalization rate decreased significantly, from 24% to 14%. Substance related hospitalizations also decreased significantly, from 5% to 3%. Average number of hospitalizations and number of days hospitalized decreased even after adjusting for sociodemographic factors. PROS serves a high number of COD patients, and the number of psychiatric and substance related hospitalizations decreased after an episode of PROS, as did the number of days hospitalized. Findings support the maintenance of psychiatric rehabilitation models that include recovery oriented components. Further analyses with control samples are proposed.


Temperature, pressure and cholesterol effects on bilayer fluidity; a comparison of pyrene excimer/monomer ratios with the steady-state fluorescence polarization of diphenylhexatriene in liposomes and microsomes.
Pyrene excimer/monomer (E/M) ratios have been compared with the steady-state fluorescence polarization (P) of diphenylhexatriene (DPH) in multilamellar liposomes of dilaurylphosphatidylcholine and rat liver microsomes. The purpose was to use the well-understood properties of DPH to reveal the nature of bilayer fluidity which pyrene manifests as an E/M ratio. Reducing the temperature (from 37 degrees C to 8 degrees C), increasing the hydrostatic pressure (from 0.1 to 70 MPa), and, in liposomes, cholesterol enrichment (up to 0.30 mole fraction) separately decreased the E/M ratios and increased P. The pyrene membrane/buffer partition coefficient was affected by temperature but not by pressure, and in the case of cholesterol enrichment, it was assumed to be unaffected. Plots of P as a function of the E/M ratio showed the two to be closely correlated (r = 0.99 in liposomes and 0.96 in microsomes), independent of the treatment used to reduce fluidity. The apparent activation volume and enthalpy for excimer formation was calculated and compared with published data. Pyrene E/M ratios probably reflect the intermolecular volume (fluidity) of the outer region of the bilayer, which is reduced by a decrease in temperature and an increase in pressure and cholesterol. DPH reports the bilayer interior, which is similarly ordered by the experimental treatments. The regional distinction between the two probes, however, accounts for the divergence of E/M ratios and P, which has been reported in membranes enriched with fluidizing fatty acids.


Rational design of in situ localization solid-state fluorescence probe for bio-imaging of intracellular endogenous cysteine.
Fluorescence detection technology has been widely concerned for its advantages of low cost, simple operation, good sensitivity, real-time and non-destructive biological imaging. However, most fluorophores emit bright fluorescence in solution, and the fluorescence decreases significantly in the high concentration or solid/aggregated state, which is called aggregation-caused quenching (ACQ). Cysteine (Cys) is an important kind of amino-acid in the field of bio-medicine, whose main function is to participate in metabolism and protein synthesis, detoxification, but intracellular cysteine concentrations (30-200 μM) are much low, and direct detection of endogenous cysteine is hampered by interference with other thiols. To solve the above problems, based on solid-state fluorophore HPQ, we for the first time prepared a novel solid-state fluorescence probe MA-HPQ, for monitoring of endogenous Cys, operated by the mechanism of excited intramolecular proton transfer (ESIPT). MeO-HPQ is completely insoluble in water, has very strong solid-state fluorescence with the maximum emission wavelength of 510 nm and the maximum excitation wavelength of 365 nm. This special property makes it very suitable for confocal microscopy compared with ordinary water-soluble fluorescent dyes. Due to the large Stokes shift (145 nm), MA-HPQ has very desirable advantages: reduced interference of background fluorescence, increased sensitivity, and enhanced contrast of biological imaging. More importantly, by preventing it from establishing internal hydrogen bonds, which is between imine nitrogen and phenolic hydroxyl groups, it can be made insoluble in water and have strong fluorescence properties, and the process is reversible. The ESIPT process can be blocked by masking phenolic hydroxyl, which can inhibit fluorescence to a large extent. In the presence of Cys, the probe reacts, releasing free MeO-HPQ, and begins to form a precipitated solid. The precipitated solid emitted bright green solid-state fluorescence, which was enhanced 43 times more than MA-HPQ. These results indicate that the probe MA-HPQ can be suitable to real spatiotemporal imaging of endogenous cysteine in HeLa cells. The excellent performance of the probe makes it applying for the visualization detection of endogenous cysteine in living cells and tissues with obtaining satisfactory results.


Erratum.
Erratum to "High-dose Dexamethasone Therapy as the Initial Treatment for Idiopathic Thrombocytopenic Purpura: Protocol for a Multicenter, Open-label, Single Arm Trial" Vol.72, No.2, pp197-201.


Treatment of premature ventricular complexes with acebutolol.
The effect of intravenous acebutolol versus saline solution on frequent premature ventricular complexes was evaluated in a double-blind, randomized study in 20 patients, including 3 with chronic obstructive pulmonary disease. Frequent premature ventricular complexes were abolished or reduced by 75% or more in none of 12 patients given saline solution but in 18 of 20 patients (90%) given acebutolol (P less than 0.001). This therapeutic effect of acebutolol persisted for at least 2.5 hours in 17 of 20 patients (85%), for at least 3.5 hours in 14 (70%) and for at least 4 hours in 8 (40%). Acebutolol was well tolerated by the three patients with chronic obstructive pulmonary disease. These data indicate that intravenous acebutolol is useful in the treatment of premature ventricular complexes.


Elements modulating the block of transcription elongation at the adenovirus 2 attenuation site.
We have previously reported that a block of transcription elongation is functioning in vivo and in vitro within the leader sequences of SV40 and the adenovirus 2 major late transcription units and in the regulation of transcription of the P4 promoter of minute virus of mice. In the present study using the HeLa whole cell extract-Sarkosyl system with adenovirus 2 major late promoter as a template we have analyzed several basic parameters that can contribute to our understanding of the mechanism that regulates the elongation block at the adenovirus 2 attenuation site. We show that the elongation block is augmented at elevated temperatures (40-45 degrees C). The elongation block can be reversed by the addition of 0.2 M NaCl to the transcription reaction and the reversibility is temperature-dependent. Furthermore, while at 30-35 degrees C the elongation block is reversible with dilution of the Sarkosyl, at 40-45 degrees C it is only partially reversible. These results may indicate that a factor(s) is involved in the regulation of the elongation block and/or that the conformation of the transcription complex is temperature dependent. Finally, we show that the extent of the elongation block is dependent on the consecutive T residues at the attenuation site and we discuss the involvement of RNA secondary structure in eliciting the elongation block.


Epidermoid cyst in an intrapancreatic accessory spleen in the pancreas head: a case report.
An epidermoid cyst in an intrapancreatic accessory spleen (ECIPAS) in the pancreas head is an extremely rare condition. The natural course of this condition is not well known, and it is difficult to diagnose before surgery due to the lack of specific imaging findings. A tumor was found in the head of the pancreas in a 68-year-old man with abdominal distension and discomfort. Magnetic resonance imaging (MRI) suggested a malignant tumor, such as a colloid cancer. The tumor was removed surgically, with pathologic examination showing that it was an ECIPAS. ECIPAS cannot be easily distinguished from other pancreatic cystic tumors, making it necessary to include ECIPAS in the differential diagnosis of these tumors. Unnecessary surgical resection may be avoided by more accurate preoperative diagnosis based on clinical and imaging characteristics.


Mental health and the veterinary profession.
Ellie Patterson, Vetlife marketing officer, summarises the services offered by Vetlife - an independent, confidential and free charity for everyone in the veterinary community.


Dopamine agonist action of phencyclidine.
Although the psychotomimetic action of phencyclidine is often used to model a hypoglutamate theory of psychosis or schizophrenia, work also exists showing that phencyclidine has a significant affinity for the dopamine D2 receptor. The present study was done to determine whether phencyclidine has a direct functional dopamine-like action on cells. The effect of phencyclidine was tested on the release of prolactin from rat cultured anterior pituitary cells. It was found that the release of prolactin was 50% inhibited by 4 nM phencyclidine. This strong dopamine-like agonist action at the functional high-affinity state of the dopamine D2 receptor by the phencyclidine psychotomimetic is consistent with the dopamine hypothesis of psychosis.


Towards a selective adsorbent for arsenate and selenite in the presence of phosphate: Assessment of adsorption efficiency, mechanism, and binary separation factors of the chitosan-copper complex.
The potential for a chitosan-copper polymer complex to select for the target contaminants in the presence of their respective competitive ions was evaluated by synthesizing chitosan-copper beads (CCB) for the treatment of (arsenate:phosphate), (selenite:phosphate), and (selenate:sulfate). Based on work by Rhazi et al., copper (II) binds to the amine moiety on the chitosan backbone as a monodentate complex (Type I) and as a bidentate complex crosslinking two polymer chains (Type II), depending on pH and copper loading. In general, the Type I complex exists alone; however, beyond threshold conditions of pH 5.5 during synthesis and a copper loading of 0.25 mol Cu(II)/mol chitosan monomer, the Type I and Type II complexes coexist. Subsequent chelation of this chitosan-copper ligand to oxyanions results in enhanced and selective adsorption of the target contaminants in complex matrices with high background ion concentrations. With differing affinities for arsenate, selenite, and phosphate, the Type I complex favors phosphate chelation while the Type II complex favors arsenate chelation due to electrostatic considerations and selenite chelation due to steric effects. No trend was exhibited for the selenate:sulfate system possibly due to the high Ksp of the corresponding copper salts. Binary separation factors, α12, were calculated for the arsenate-phosphate and selenite-phosphate systems, supporting the mechanistic hypothesis. While, further research is needed to develop a synthesis method for the independent formation of the Type II complexes to select for target contaminants in complex matrices, this work can provide initial steps in the development of a selective adsorbent.


Excretion of biogenic amine metabolites in anorexia nervosa.
Urinary metabolites of catecholamines and indoleamines have been investigated in 16 patients with anorexia nervosa (AN) and 13 controls using a HPLC-method. Vanillic mandelic acid, 3-methoxy-4-hydroxyphenylglycol, homovanillic acid, 3,4-dihydroxyphenylacetic acid, 5-hydroxyindole acetic acid and indoleacetic acid were significantly decreased in the pre-treatment phase. In four patients long-term treatment including parenteral and enteral nutrition together with psychological methods resulted in an increase in the levels of these substances and this correlated with increased weight gain and urinary creatinine. It is concluded that both central and peripheral disturbances are involved in AN, particularly with regard to biogenic amine metabolism.


Association between Gamma-Glutamyltransferase Level and Risk of Stroke: A Systematic Review and Meta-analysis of Prospective Studies.
Stroke is often regulated by a number of modifiable and nonmodifiable risk factors. Recently, studies suggested high gamma-glutamyltransferase (GGT) level may be associated with stroke, but drew inconsistent conclusions. So, we conducted a meta-analysis to evaluate the relationship between GGT level and risk of stroke. We systematically searched PubMed, Embase, and Cochrane Library (updated to January 2015) for prospective cohort studies. Then, relative risk (RR) with 95% confidence interval (CI) was used to assess the association. Regression analyses, subgroup analyses, and sensitivity analyses were also performed. The Begg test, Egger test, and the trim-and-fill method were used to assess potential publication bias. A total of 5707 cases and 926,497 participants in 10 prospective studies were included. Overall, high GGT level has a positive association with increased risk of stroke (RR = 1.28; 95% CI, 1.16-1.43). In the subgroup analyses, a positive association was consistently observed in each subgroup except in the women subgroup (RR = 1.45; 95% CI, .9-2.34) and a large number of stroke events subgroups (≥ 500) (RR = 1.25; 95% CI, .85-1.84). Heterogeneity was significantly reduced in the subgroup analysis by population characteristics. In the publication bias test, the resulting adjusted RR remained significant (RR = 1.10; 95% CI, 1.00-1.21) after using the trim-and-fill method. Our meta-analysis provides evidence that a high level of GGT is significantly associated with increased risk of stroke independently of alcohol intake. Gender and ethnicity variations may exist in the relationship between high GGT level and risk of stroke.


Deeply eosinophilic cell variant of signet-ring type of gastric carcinoma: a diagnostic dilemma.
Signet-ring cell type of gastric carcinoma can easily escape detection by the pathologist, and this confusion is compounded by the presence of morphologic patterns other than the conventional appearance. One such morphologic variant is composed of deeply eosinophilic cells containing minute cytoplasmic granules with neutral mucin. With this morphology, epithelioid gastrointestinal stromal tumor (GIST), and hepatoid and rhabdoid variants of adenocarcinoma enter the list of diagnostic possibilities, and accurate diagnosis is essential for therapeutic and prognostic considerations. We report a 28-year-old man who presented with dyspepsia, nausea, vomiting, and black-colored stools. Endoscopy revealed a growth at the incisura, a biopsy specimen from which was reported as a poorly differentiated tumor. The patient underwent subtotal gastrectomy and the resected specimen showed a 6.5 x 4-cm circumferential growth in the antropyloric region. Histological sections from the growth showed transmural infiltration by deeply eosinophilic cells with hyperchromatic nuclei. A differential diagnosis of epithelioid GIST, rhabdoid variant, hepatoid variant, and signet-ring cell adenocarcinoma was considered. Special stains and immunohistochemistry confirmed the tumor to be a variant of signet-ring cell adenocarcinoma of the stomach. Morphologic variations of signet-ring cell carcinoma pose an important diagnostic dilemma, which has therapeutic and prognostic significance for the patient. These variants need to be recognized by pathologists to enable an accurate diagnosis.


Assessment of Oxidative Stress in Patients with Premature Canities.
Premature canities is a common, yet unexplored disorder. Oxidative stress levels have been evaluated within the greying hair follicle but not in the sera of patients with premature canities. To evaluate the oxidative stress parameters in the sera of patients with premature canities. A pilot case-controlled study, conducted in a tertiary care setup in Delhi during November 2011 to December 2012. Fifty-two self-reporting cases of premature canities (age of onset <20 years) and 30 healthy controls were recruited from outpatient Department of Dermatology. Oxidative stress parameters (serum malonaldehyde (MDA), whole blood reduced glutathione (rGSH) and serum ferric reducing antioxidant potential [FRAP]) were assessed in cases and controls. Mann-Whitney test was used to compare the oxidative stress parameters between the two groups (SPSS version 17.0, SPSS Inc, Chicago, USA; P < 0.05 considered as significant). The age and sex distribution of cases and controls was comparable. The mean serum levels of MDA were higher in cases than controls (3.7 ± 1.6 nmol/ml vs. 2.8 ± 1.5 nmol/ml; P = 0.01). The GSH levels were lower in the cases than controls (31.5 ± 8.9 mg/dl vs. 36.6 ± 16.9 mg/dl; P = 0.064). Similarly, the mean FRAP levels were lower in the cases than controls (400 ± 70 nmol/ml vs. 430 ± 80 nmol/ml; P = 0.038). Patients with premature canities had a higher level of pro-oxidants and lower levels of antioxidants than controls. This is the first humble attempt to document the oxidative stress parameters in sera of patients with premature canities, further studies with larger sample size are required to reach a definite conclusion.


Common variable immunodeficiency disorder (CVID)-related liver disease: assessment of the main histological aspects using novel semiquantitative scoring systems, image analysis and correlation with clinical parameters of liver stiffness and portal hypertension.
We aimed to investigate the relationship between T-cell-mediated sinusoidal injury, nodular regenerative hyperplasia like changes (NRH-LC) and fibrosis, clinical measures of fibrosis and portal hypertension, and progression rate in common variable immunodeficiency disorder (CVID)-related liver disease. This is a retrospective single-centre study. Liver biopsies from CVID patients with liver disease were reviewed to assess for NRH-LC, fibrosis and elastosis, including collagen and elastin proportionate areas. CD3 positive T-cells infiltration and sinusoidal endothelial changes by CD34 expression were quantified by image analysis and a semiquantitative method, respectively. These findings were correlated with liver stiffness measurements (LSM) and hepatic venous pressure gradient (HVPG). NRH-LC and pericellular elastosis were present in most biopsies (32/40 and 38/40, respectively). All biopsies showed fibrosis, which was limited to pericellular in 21/40 (52.5%) and included bridging fibrous septa in 19/40 (47.5%). 28/40 liver biopsies showed enhanced sinusoidal expression of CD34. There were more CD3 positive cells in biopsies with NRH-LC compared with those without. There was no significant correlation between LSM, HVPG and fibrosis/elastosis scores. Five of seven patients with at least two biopsies showed progression in fibrosis stage. NRH-LC and fibrosis in CVID patients often coexist along with the presence of sinusoidal endothelial changes and sinusoidal lymphocytic infiltration. Fibrosis progresses over time, and significant fibrosis can be observed in young patients (<30 years old), potentially reflecting a more aggressive form of CVID-related liver disease. Further studies are necessary to investigate the relationship between histological findings, clinical measures of fibrosis and portal hypertension and outcome.


Natural photoswitches expose STIM1 activation steps.
STIM1, an ER-located Ca2+ sensor, activates Orai1 channels upon Ca2+-storedepletion. Prior to this, STIM1 undergoes a sequence of conformational changes, which cannot be controlled individually with high spatiotemporal resolution. Ma et al. [1] used the power of optogenetic engineering to transfer light-sensitivity to STIM1 and precisely characterize individual STIM1 activation steps.


Gene Therapy of the Peripheral Nervous System: The Enteric Nervous System.
The enteric nervous system (ENS) is a complex network of ganglia embedded in the walls of the gastrointestinal (GI) tract. The highly integrated enteric neural network can act independently of sympathetic and parasympathetic innervation to control all aspects of GI function. Due to the central role the ENS plays in GI physiology, the ability to manipulate genetic expression within the ENS is an invaluable tool for both research and clinical applications. Here, we describe a method for gene delivery to the ENS using direct injections of adeno-associated virus (AAV) into the gut wall. This method is able to achieve transgene expression in both neurons and glia located in discrete areas of the ENS.


A new administration form of ritodrine. Ritodrine plasma levels with sustained-release capsules in a steady state.
A sustained-release administration form of oral ritodrine was introduced to cope with the problem of the short dosage interval inherent to conventional oral ritodrine administration. The bioavailability of this application form was assessed in a clinical study including 11 patients. With a dosage frequency of only 3 times daily and a dosage form index of 1.8, plasma levels of 14.6-26.5 ng/ml ritodrine (equivalent to an infusion rate of approximately 50 micrograms/min) were obtained without serious cardiovascular or metabolic side effects. It was concluded that with this new formulation clinical applicability of orally administered ritodrine in tocolytic therapy has increased.


General practitioners' evaluations of optimal timing to initiate advance care planning for patients with cancer, organ failure, or multimorbidity: A health records survey study.
Appropriate timing to initiate advance care planning is difficult, especially for individuals with non-malignant disease in community settings. To identify the optimal moment for, and reasons to initiate advance care planning in different illness trajectories. A health records survey study; health records were presented to 83 GPs with request to indicate and substantiate what they considered optimal advance care planning timing within the 2 years before death. We used quantitative and qualitative analyses. We selected and anonymized 90 health records of patients who died with cancer, organ failure or multimorbidity, from a regional primary care registration database in the Netherlands. The median optimal advance care planning timing according to the GPs was 228 days before death (interquartile range 392). This moment was closer to death for cancer (87.5 days before death, IQR 302) than for organ failure (266 days before death, IQR 401) and multimorbidity (290 days before death, IQR 389) (p < 0.001). The most frequently mentioned reason for cancer was "receiving a diagnosis" (21.5%), for organ failure it was "after a period of illness" (14.7%), and for multimorbidity it was "age" and "patients" expressed wishes or reflections' (both 12.0%). The optimal advance care planning timing and reasons to initiate advance care planning indicated by GPs differ between patients with cancer and other illnesses, and they also differ between GPs. This suggests that "the" optimal timing for ACP should be seen as a "window of opportunity" for the different disease trajectories.


Exploring the pathways of homologous recombination.
There has been significant progress in elucidating the mechanisms by which meiotic and mitotic recombination occur. Double-strand breaks in particular have been the object of attention in studies on meiotic gene conversion, site-specific mitotic recombination, the repair of transposon excision and the transformation of cells with linearized DNA. A combination of genetic analysis and physical studies of molecular recombination intermediates have established that double-strand breaks can occur by two different mechanisms.


Multivariate morphological brain signatures enable individualized prediction of dispositional need for closure.
Need for closure (NFC) reflects stable individual differences in the desire for a quick, definite, and stable answer to a question. A large body of research has documented the association between NFC and various cognitive, emotional and social processes. Despite considerable interest in psychology, little effort has been made to uncover the neural substrates of individual variations in NFC. Herein, we took a data-driven approach to predict NFC trait combining machine learning framework and the whole-brain grey matter volume (GMV) features, which represent a reliable brain imaging measure and have been commonly employed to explore neural basis underlying individual differences of cognition and behaviors. Brain regions contributing to the prediction were then subjected to functional connectivity and decoding analyses for a quantitative inference on their psychophysiological functions. Our results indicated that multivariate patterns of GMV derived from multiple regions across distributed brain systems predicted NFC at individual level. The contributing regions are distributed across the emotional processing network (e.g., striatum), cognitive control network (e.g., lateral prefrontal cortex), social cognition network (e.g., temporoparietal junction) and perceptual processing network (e.g., occipital cortex). The current study provided the first evidence that dispositional NFC is embodied in multiple large-scale brain networks, helping to delineate a more complete picture about the neuropsychological processes that support individual differences in NFC. Beyond these findings, the current interdisciplinary approach to constructing and interpreting neuroimaging-based prediction model of personality traits would be informative to a wide range of future studies on personality.


Effects of adrenergic agents on rat peritoneal macrophages activated in vitro by acetylated low-density lipoprotein.
Macrophages interact with modified lipoproteins and alter their functional status. In this study, the effects of the adrenergic agents adrenaline, isoproterenol, and dobutamine on macrophages activated with acetylated low-density lipoprotein were tested. The aim of this investigation was to determine whether adrenergic agents influence biologically significant functions of these cells in an in vitro model of macrophage-lipoprotein acute interaction. Rat peritoneal macrophages were incubated with acetylated low-density lipoprotein for 16 h, with or without adrenergic agents. Hydrogen peroxide and nitric oxide production and acid phosphatase activities in the supernatant and cell lysate were assayed. Adrenaline and isoproterenol inhibited the production of hydrogen peroxide, stimulated the production of nitric oxide, and increased the extracellular activity of acid phosphatase in the lipoprotein-activated cells. Dobutamine increased the extracellular, but decreased the intracellular acid phosphatase activity. Adrenaline and dobutamine also produced changes in the cell size and nuclear morphology of the macrophages. Macrophages are activated in vitro by acetylated low-density lipoprotein, and their functions and morphology are modified under the influence of adrenergic agents. Certain effects could be attributed to changes in cyclic AMP levels.


Fibroblast growth factor 21: a novel long-acting hypoglycemic drug for canine diabetes.
Currently, insulin is commonly used in the clinical management of canine diabetes. However, it must be injected preprandially causing much inconvenience to the owners. Therefore, the development of long-acting hypoglycemic agents has attracted much attention in the scientific community. This study aimed to investigate the long-acting hypoglycemic effect of canine fibroblast growth factor 21 (cFGF-21) in diabetic dogs. Diabetic dogs were administered with cFGF-21, polyethylene glycol-modified cFGF-21 (PEG-cFGF-21), or insulin once a day, once every 2, 3, or 4 days subcutaneously. The results showed that cFGF-21 and PEG-cFGF-21 maintained blood glucose comparable to normal levels for 2 and 3 days respectively while insulin maintained the blood glucose for only 2 h after a single injection. After treatment with cFGF-21, oral glucose tolerance test (OGTT) was significantly improved with glycosylated hemoglobin (HbA1c) close to the normal levels. In addition, cFGF-21 significantly repaired islet β cells, increased insulin content, and protected the pancreas from streptozotocin-induced injury. Furthermore, cFGF-21 exhibited both antioxidant and anti-inflammatory properties in the pancreas. We conclude, therefore, that cFGF-21 and PEG-cFGF-21 can maintain blood glucose comparable to normal levels for 2 and 3 days respectively after a single dose. The long-acting efficacy of cFGF-21 can be attributed to improvement in oxidative stress and the reduction of inflammation in the pancreas.


Use of sera from humans and dolphins with lacaziosis and sera from experimentally infected mice for Western Blot analyses of Lacazia loboi antigens.
Antibodies in the sera of patients with lacaziosis recognized an approximately 193-kDa antigen and other Lacazia loboi antigens. Paracoccidioides brasiliensis gp43 antigen was detected by all evaluated sera, but they failed to detect a protein with the same molecular mass in L. loboi extracts. This study is the first to examine the humoral response to L. loboi antigens by using multiple host sera.


Rational design of biodegradable thermoplastic polyurethanes for tissue repair.
As a type of elastomeric polymers, non-degradable polyurethanes (PUs) have a long history of being used in clinics, whereas biodegradable PUs have been developed in recent decades, primarily for tissue repair and regeneration. Biodegradable thermoplastic (linear) PUs are soft and elastic polymeric biomaterials with high mechanical strength, which mimics the mechanical properties of soft and elastic tissues. Therefore, biodegradable thermoplastic polyurethanes are promising scaffolding materials for soft and elastic tissue repair and regeneration. Generally, PUs are synthesized by linking three types of changeable blocks: diisocyanates, diols, and chain extenders. Alternating the combination of these three blocks can finely tailor the physio-chemical properties and generate new functional PUs. These PUs have excellent processing flexibilities and can be fabricated into three-dimensional (3D) constructs using conventional and/or advanced technologies, which is a great advantage compared with cross-linked thermoset elastomers. Additionally, they can be combined with biomolecules to incorporate desired bioactivities to broaden their biomedical applications. In this review, we comprehensively summarized the synthesis, structures, and properties of biodegradable thermoplastic PUs, and introduced their multiple applications in tissue repair and regeneration. A whole picture of their design and applications along with discussions and perspectives of future directions would provide theoretical and technical supports to inspire new PU development and novel applications.


Economic evaluation of cell salvage in pediatric surgery.
Red blood cells are a scarce resource with demand outstripping supply. Use of intraoperative red cell salvage (CS) - the process of collecting shed blood during surgery and reinfusing it to patients - is often used as an effective blood conservation strategy. However, little is known about the economic impact of CS during pediatric surgery. A decision tree model was used to estimate the transfusion-related costs per patient (2010 USD) from a healthcare system perspective of four transfusion strategies among children undergoing elective orthopedic or cardiac surgery: (i) CS followed by allogeneic transfusion, (ii) CS followed by autologous transfusion, (iii) allogeneic transfusion alone, and (iv) autologous transfusion alone. Cell salvage and allogeneic transfusion was the least expensive strategy (USD 883.3) followed by CS and autologous blood transfusion (USD 1,269.7), allogeneic transfusion alone (USD 1,443.0), and autologous transfusion alone (USD 1,824.7). Savings associated with CS use persisted in separate analyses of orthopedic and cardiac surgery, as well as in one-way and probabilistic sensitivity analyses. Use of CS, particularly along with allogeneic blood transfusion, appears cost-saving and cost-effective in pediatric surgery.


BRAF activation by metabolic stress promotes glycolysis sensitizing NRASQ61-mutated melanomas to targeted therapy.
NRAS-mutated melanoma lacks a specific line of treatment. Metabolic reprogramming is considered a novel target to control cancer; however, NRAS-oncogene contribution to this cancer hallmark is mostly unknown. Here, we show that NRASQ61-mutated melanomas specific metabolic settings mediate cell sensitivity to sorafenib upon metabolic stress. Mechanistically, these cells are dependent on glucose metabolism, in which glucose deprivation promotes a switch from CRAF to BRAF signaling. This scenario contributes to cell survival and sustains glucose metabolism through BRAF-mediated phosphorylation of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-2/3 (PFKFB2/PFKFB3). In turn, this favors the allosteric activation of phosphofructokinase-1 (PFK1), generating a feedback loop that couples glycolytic flux and the RAS signaling pathway. An in vivo treatment of NRASQ61 mutant melanomas, including patient-derived xenografts, with 2-deoxy-D-glucose (2-DG) and sorafenib effectively inhibits tumor growth. Thus, we provide evidence for NRAS-oncogene contributions to metabolic rewiring and a proof-of-principle for the treatment of NRASQ61-mutated melanoma combining metabolic stress (glycolysis inhibitors) and previously approved drugs, such as sorafenib.


Pension-Spiking, Free-Riding, and the Effects of Pension Reform on Teachers' Earnings.
In many states, local school districts are responsible for setting the earnings that determines the size of pensions, but are not required to make contributions to cover the resulting state pension fund liabilities. In this paper, I document evidence that this intergovernmental incentive inherent in public sector defined benefit pension systems distorts the amount and timing of income for public school teachers. I use the introduction of a policy that required experience-rating on earnings increases above a certain limit in a differences-in-differences framework to identify whether districts are willing to pay the full costs of their earnings promises. Because of the design of the policy, overall earnings of teachers near retirement did not change. Instead, districts that previously provided one-time pay increases shifted to smaller increments spread out over several years. In addition, some districts that did not practice pension-spiking prior to the reform appear to begin providing payments up to the new, lower limit, perhaps due to increased salience of the fiscal incentive. Therefore, the policy was ineffective at decreasing pension costs.