Benzyl isothiocyanate promotes miR-99a expression through ERK/AP-1-dependent pathway in bladder cancer cells.
Benzyl isothiocyanate (BITC), a bioactive natural product present in cruciferous vegetables, has been proved to prevent cancer progression through various mechanisms. In our previous report, we proved that BITC exhibits antitumor effects in bladder cancer by suppressing IGF1R, FGFR3, and mTOR, which is mediated by miR-99a expression. In this study, we identified the signal pathway involved in regulating miR-99a expression after BITC exposure in bladder cancer. Treatment with different BITC concentrations resulted in induction of miR-99a expression in bladder cancer cell lines. Activation of extracellular signal-regulated protein kinase (ERK) and c-jun N-terminal kinase was observed in bladder cancer after BITC treatment for 24 hours. Interestingly, by using a chemical inhibitor of candidate pathways, we found that only the ERK signal pathway is required for miR-99a expression. Furthermore, we evaluated the transcription factor that may contribute to miR-99a expression in response to BITC treatment. The results indicated that c-Jun/AP-1 was activated after BITC treatment. Moreover, we confirmed c-Jun/AP-1 activation through immunofluorescence and the luciferase reporter assay. The results showed that BITC treatment markedly improved nuclear translocation of c-Jun/AP-1 and luciferase activity dose dependently. Finally, pretreatment with the ERK inhibitor U0126 diminished c-Jun phosphorylation and transcriptional activation, suggesting that BITC elicits ERK/c-Jun signal transduction, which is responsible for miR-99a expression in bladder cancer. The present work identifies the mechanism involved in upregulation miR-99a after BITC treatment, which provides an explanation for BITC biological function in our previous work.


Material-specific thrombin generation following contact between metal surfaces and whole blood.
Little is known about the blood compatibility of metals used in various medical devices. We have previously shown that titanium and derivatives thereof are among the most thrombogenic materials which may explain its outstanding osteointegrating properties. The aim of the present study was to characterize the thrombogenic and complement-activating properties of various metals used today in medical applications. Polyester chips were coated with 50- to 100-nm thick layers of aluminium, iridium, indium, nickel, tantalum, tin, titanium, or zirconium using magnetron sputtering. The metal-coated chips were then incubated in direct contact with whole blood in an in vitro chamber model, and the blood was then analyzed for platelet counts, thrombin-antithrombin (AT), fXIIa-AT, fXIa-AT and fXIIa-C1INH complexes and the complement parameters C3a and sC5b-9. Titanium, tantalum and indium were found to exhibit pronounced thrombogenic properties, whereas aluminium, nickel and, in particular, iridium were essentially non-thrombogenic. Tin and zirconium were intermediate activators. All metals activated complement to a similar degree, with the exception of aluminium, which had more pronounced activating properties. This study clearly indicates that metals indeed have varying thrombogenic and complement activating properties. These studies have implications for the selection of metals intended for medical applications.


Identification of sequence elements that confer cell-type-specific control of MF alpha 1 expression in Saccharomyces cerevisiae.
The MF alpha 1 gene of Saccharomyces cerevisiae, a major structural gene for mating pheromone alpha factor, is an alpha-specific gene whose expression is regulated by the mating-type locus. To study the role of sequences upstream of MF alpha 1 in its expression and regulation, we generated two sets of promoter deletions: upstream deletions and internal deletions. By analyzing these deletions, we have identified a TATA box and two closely related, tandemly arranged upstream activation sites as necessary elements for MF alpha 1 expression. Two upstream activation sites were located ca. 300 and 250 base pairs upstream of the MF alpha 1 transcription start points, which were also determined in this study. Each site contained a homologous 22-base-pair sequence, and both sites were required for maximum transcription level. The distance between the upstream activation sites and the transcription start points could be altered without causing loss of transcription efficiency, and the sites were active in either orientation with respect to the coding region. These elements conferred cell type-specific expression on a heterologous promoter. Analysis with host mating-type locus mutants indicates that these sequences are the sites through which the MAT alpha 1 product exerts its action to activate the MF alpha 1 gene. Homologous sequences with these elements were found in other alpha-specific genes, MF alpha 2 and STE3, and may mediate activation of this set of genes by MAT alpha 1.


MGAT5/TMEM163 variant is associated with prognosis in ursodeoxycholic acid-treated patients with primary biliary cholangitis.
Primary biliary cholangitis (PBC) is a chronic immune-mediated liver disease. Previous genome-wide meta-analysis has identified the association between variants in TMEM163 with PBC. Here we aimed to evaluate the association between variants near the reported risk loci of TMEM163 at 2q21.3 and prognosis of PBC patients. We performed a retrospective analysis of 347 PBC patients treated with ursodeoxycholic acid (UDCA) for at least 1 year. We collected clinical data at diagnosis and 1 year after UDCA treatment. SNPs within 200 kb upstream and downstream of the lead variant were genotyped and screened. We identified that rs661899 near MGAT5 and TMEM163 showed the strongest association with prognosis in PBC patients. Patients carrying the rs661899 T allele tended to respond incompletely to UDCA treatment and had worse performances in laboratory values including aspartate aminotransferase (53.5 vs 32 vs 28.5 U/L, p = 0.001), alkaline phosphate (157.25 vs 125 vs 113 U/L, p = 0.001), albumin (41.5 vs 42.3 vs 43.7 g/L, p = 0.008) and bilirubin (19.2 vs 14.9 vs 12.85 μmol/L, p = 0.001). GLOBE scores (p = 4.8 × 10-5) and UK-PBC risk scores (p = 4.6 × 10-4) were strongly correlated with rs661899 genotype. Patients with TT genotype had a higher risk for adverse events compared with CC genotype (p = 0.039) during the 1-year follow-up. Results were also verified in an independent cohort. PBC patients carrying the rs661899 T allele are associated with poor prognosis and adverse outcomes after 1-year UDCA therapy.


The G protein-coupled oestrogen receptor 1 agonist G-1 disrupts endothelial cell microtubule structure in a receptor-independent manner.
The G protein-coupled oestrogen receptor GPER1, also known as GPR30, has been implicated in oestrogen signalling, but the physiological importance of GPER1 is not fully understood. The GPER1 agonist G-1 has become an important tool to assess GPER1-mediated cellular effects. Here, we report that this substance, besides acting via GPER1, affects the microtubule network in endothelial cells. Treatment with G-1 (3 μM) for 24 h reduced DNA synthesis by about 60 % in mouse microvascular endothelial bEnd.3 cells. Treatment with 3 μM G-1 prevented outgrowth of primary endothelial cells from mouse aortic explants embedded in Matrigel. Treatment with G-1 (0.3-3 μM) for 24 h disrupted bEnd.3 cell and HUVEC microtubule structure in a concentration-dependent manner as assessed by laser-scanning confocal immunofluorescence microscopy. G-1-induced (3 μM) disruption of microtubule was observed also after acute (3 and 6 h) treatment and in the presence of the protein synthesis inhibitor cycloheximide. Disruption of microtubules by 3 μM G-1 was observed in aortic smooth muscle cells obtained from both GPER1 knockout and wild-type mice, suggesting that G-1 influences microtubules through a mechanism independent of GPER1. G-1 dose dependently (10-50 μM) stimulated microtubule assembly in vitro. On the other hand, microtubules appeared normal in the presence of 10-50 μM G-1 as determined by electron microscopy. We suggest that G-1-promoted endothelial cell anti-proliferation is due in part to alteration of microtubule organization through a mechanism independent of GPER1. This G-1-promoted mechanism may be used to block unwanted endothelial cell proliferation and angiogenesis such as that observed in, e.g. cancer.


Corneal Breakthrough Haze After Photorefractive Keratectomy With Mitomycin C: Incidence and Risk Factors.
To identify preoperative and intraoperative factors affecting breakthrough corneal haze incidence after photorefractive keratectomy (PRK) with mitomycin C (MMC). In this retrospective study of PRK performed at the Care Vision Refractive Laser Center, Tel Aviv, Israel, a total of 7535 eyes (n = 3854 patients; mean age ± SD, 26 ± 6 years; 55% men) underwent PRK with intraoperative MMC application. Patients with histories of corneal pathology or surgery were excluded. Incidence, time of onset, and corneal haze severity were documented on follow-up of 118 ± 110 days. Eyes were grouped by preoperative refraction [low (≤-3D), moderate (-3D to -6D), or high (>-6D) myopia; low (≤3D) or high (>3D) astigmatism; low or high hyperopia]; by intraoperative time (above or below 40 seconds); and by MMC application time (above or below 40 seconds). The main outcome measures were incidence, onset time, and severity of corneal haze. The haze incidence was 2.1% in eyes with high myopia versus 1.1% in those with low to moderate myopia (P = 0.002), and 3.5 times higher in eyes with high than with low astigmatism (P < 0.05). The overall incidence was higher in eyes treated for hyperopia (10.8%) than for myopia (1.3%) (P = 0.0001). In eyes with moderate myopia, the haze incidence was lower in MMC application time ≥40 seconds (0%) than in <40 seconds (1.3%) (P = 0.03). After surgery, a mild early haze incidence peaked at 68.8 ± 6 days and severe late haze at 115 ± 17 days (P = 0.02). Hyperopic and large myopic or astigmatic corrections carry higher risk of haze. Longer MMC application might have beneficial haze prevention.


Novel and unusual genes for nitrogen and metal cycling in Planctomycetota- and KSB1-affiliated metagenome-assembled genomes reconstructed from a marine subsea tunnel.
The Oslofjord subsea road tunnel is a unique environment in which the typically anoxic marine deep subsurface is exposed to oxygen. Concrete biodeterioration and steel corrosion in the tunnel have been linked to the growth of iron- and manganese-oxidizing biofilms in areas of saline water seepage. Surprisingly, previous 16S rRNA gene surveys of biofilm samples revealed microbial communities dominated by sequences affiliated with nitrogen-cycling microorganisms. This study aimed to identify microbial genomes with metabolic potential for novel nitrogen- and metal-cycling reactions, representing biofilm microorganisms that could link these cycles and play a role in concrete biodeterioration. We reconstructed 33 abundant, novel metagenome-assembled genomes (MAGs) affiliated with the phylum Planctomycetota and the candidate phylum KSB1. We identified novel and unusual genes and gene clusters in these MAGs related to anaerobic ammonium oxidation, nitrite oxidation, and other nitrogen-cycling reactions. Additionally, 26 of 33 MAGs also had the potential for iron, manganese, and arsenite cycling, suggesting that bacteria represented by these genomes might couple these reactions. Our results expand the diversity of microorganisms putatively involved in nitrogen and metal cycling, and contribute to our understanding of potential biofilm impacts on built infrastructure.


Reference interval studies: what is the maximum number of samples recommended?
Little attention has been paid to the maximum number of specimens for reference interval calculation, i.e., the number of specimens beyond which there is no further benefit in reference interval calculation. We present a model for the estimation of the maximum number of specimens for reference interval studies based on setting the 90% confidence interval of the reference limits to be equal to the analyte reporting interval. Equations describing the bounds on the upper and lower 90% confidence intervals for logarithmically transformed and untransformed data were derived and applied to determine the maximum number of specimens required to calculate a reference interval for 12 common chemistry and hematology analytes. Maximum sample sizes ranged from 126 to 18,171 and depended on the standard deviation of the population, any transformation involved and on the chosen reporting interval. This paper demonstrates the importance of the influence of reporting interval on reference intervals. Using this technique can reduce the cost of determining a reference interval by identifying the maximum number of specimens required.


Morphological correlates of hearing loss after cochlear implantation and electro-acoustic stimulation in a hearing-impaired Guinea pig model.
Hybrid or electro-acoustic stimulation (EAS) cochlear implants (CIs) are designed to provide high-frequency electric hearing together with residual low-frequency acoustic hearing. However, 30-50% of EAS CI recipients lose residual hearing after implantation. The objective of this study was to determine the mechanisms of EAS-induced hearing loss in an animal model with high-frequency hearing loss. Guinea pigs were exposed to 24 h of noise (12-24 kHz at 116 dB) to induce a high-frequency hearing loss. After recovery, two groups of animals were implanted (n = 6 per group), with one group receiving chronic acoustic and electric stimulation for 10 weeks, and the other group receiving no stimulation during this time frame. A third group (n = 6) was not implanted, but received chronic acoustic stimulation. Auditory brainstem responses were recorded biweekly to monitor changes in hearing. The organ of Corti was immunolabeled with phalloidin, anti-CtBP2, and anti-GluR2 to quantify hair cells, ribbons and post-synaptic receptors. The lateral wall was immunolabeled with phalloidin and lectin to quantify stria vascularis capillary diameters. Bimodal or trimodal diameter distributions were observed; the number and location of peaks were objectively determined using the Aikake Information Criterion and Expectation Maximization algorithm. Noise exposure led to immediate hearing loss at 16-32 kHz for all groups. Cochlear implantation led to additional hearing loss at 4-8 kHz; this hearing loss was negatively and positively correlated with minimum and maximum peaks of the bimodal or trimodal distributions of stria vascularis capillary diameters, respectively. After chronic stimulation, no significant group changes in thresholds were seen; however, elevated thresholds at 1 kHz in implanted, stimulated animals were significantly correlated with decreased presynaptic ribbon and postsynaptic receptor counts. Inner and outer hair cell counts did not differ between groups and were not correlated with threshold shifts at any frequency. As in the previous study in a normal-hearing model, stria vascularis capillary changes were associated with immediate hearing loss after implantation, while little to no hair cell loss was observed even in cochlear regions with threshold shifts as large as 40-50 dB. These findings again support a role of lateral wall blood flow changes, rather than hair cell loss, in hearing loss after surgical trauma, and implicate the endocochlear potential as a factor in implantation-induced hearing loss. Further, the analysis of the hair cell ribbons and post-synaptic receptors suggest that delayed hearing loss may be linked to synapse or peripheral nerve loss due to stimulation excitotoxicity or inflammation. Further research is needed to separate these potential mechanisms of delayed hearing loss.


Morphometry by computerized three-dimensional reconstruction of the human carpal bones during embryogenesis.
Carpal skeleton shows drastic developmental changes during embryogenesis. At this stage, the cartilaginous matrices appear and later form models of the limb bones. The purpose of this study was to investigate the morphometry of carpal bones in humans during embryological development. We obtained digitalized histological serial sections of 18 human embryos and early fetuses from the Institute of Anatomy in Paris. Surfdriver and MSC.Patran software were used for three-dimensional reconstruction and morphometry. There was a strong correlation between the volume of the carpal cartilaginous structure and the size of the embryos (P<0.001) and an exponential correlation between the carpal volume and the percentage of volume presented by the proximal carpal row (P=0.005). According to inertia parameters, the geometry of carpal cartilaginous structure, initially plane, becomes curved during embryogenesis. Carpal bones growth follows non-homothetic transformation. The innovations in embryo reconstruction serve as new tool for scientific investigation. A hypothesis of carpal development is proposed.


Radiosensitization of hormone-refractory prostate cancer cells by gossypol treatment.
many drugs have been tested to increase the sensitivity of prostate cancer cells to radiotherapy. Gossypol, a natural polyphenolic compound extracted from the cotton plant, is one of the agents the efficacy of which has been investigated in the treatment of prostate cancer for this purpose. The main aim of this study was to investigate the best gossypol application with irradiation, when gossypol was applied either sequentially (24 h before and after irradiation) or concurrently in PC-3 hormone-refractory and radioresistant prostate cancer cells. The XTT viability assay was used to evaluate the cytotoxicity of different concentrations of gossypol in PC- 3 cells. Irradiation was applied to PC-3 cells via 6 MV photon linear accelerator and delivered 24 h before, 24 h after radiation or at the same time with gossypol administration. gossypol caused radiosensitization of PC-3 cells that are known to be radioresistant, with high Bcl-2 levels. Among different applications of gossypol and irradiation (before, after and concurrent) in prostate cancer cells, the best results were observed by the application of gossypol 24 h before irradiation. our study suggests that gossypol represents a promising novel anticancer treatment for radiosensitization of human hormone-refractory prostate cancer cells.


Efficacy of halofuginone lactate in the prevention of cryptosporidiosis in dairy calves.
To evaluate the efficacy of halofuginone lactate (Halocur; Intervet) when used prophylactically against cryptosporidiosis in dairy calves, 513 Holstein heifer calves were randomly distributed between two experimental groups. Those in the treatment group (n=259) received 0.1 mg halofuginone base per kg bodyweight daily in a 10 ml solution, for the first seven days following birth, while those in the control group (n=254) received an equal volume of a placebo solution. Faecal samples were collected weekly from all calves until three weeks of age to test for Cryptosporidium oocysts and to assess the diarrhoeal status. Growth parameters of the calves were also recorded at approximately four months of age. Calves that received halofuginone were significantly less likely to shed Cryptosporidium oocysts during the course of the study than placebo-treated calves (odds ratio 0.6, 95 per cent confidence interval 0.4 to 0.9, P=0.009). Mean growth measurements of calves in the treated group were consistently higher than those of the control calves (hipometer-derived P=0.052, wither height P=0.097) and mortality was lower in the treated group (P=0.07). However, these differences were not statistically significant (P>0.05). No association was found between the prophylactic use of halofuginone and the incidence of calf diarrhoea.


Reinforced sternal closure: the bilateral straight longitudinal wire technique.
Sternal dehiscence and mediastinitis are among the most severe complications of median sternotomy. A simplified technique of reinforced closure is described. A straight wire is inserted longitudinally on each side of the sternum, placed within the transverse wires when the latter are approximated. Using this technique in 112 patients with a precarious sternum, no cases of sternal dehiscence or mediastinitis have been seen.


Preparing radiology staff to meet service goals: a training model.
This article describes a model used to train radiology staff in customer service relations at a large southeastern medical center. Information about the needs of the radiology department and staff was acquired through quantitative and qualitative assessments. The primary goal of the training was twofold: 1) to develop employee awareness of customer expectations and 2) to develop problem-solving skills to respond to customer service related issues. Instructional methods compatible with adult learning were used and training results were assessed. Positive changes in employee attitudes and behaviors are described and recommendations for training development and implementation are discussed.


Hi-trees and their layout.
We introduce hi-trees, a new visual representation for hierarchical data in which, depending on the kind of parent node, the child relationship is represented using either containment or links. We give a drawing convention for hi-trees based on the standard layered drawing convention for rooted trees, then show how to extend standard bottom-up tree layout algorithms to draw hi-trees in this convention. We also explore a number of other more compact layout styles for layout of larger hi-trees and give algorithms for computing these. Finally, we describe two applications of hi-trees: argument mapping and business decision support.


Pediatric Posterior Infectious Uveitis.
To describe the most important cause of infectious posterior uveitis in pediatric patients. Review of the literature. The most important causes of infectious uveitis in pediatric patients are: cat-scratch disease, toxocariasis, tuberculosis, viral diseases and toxoplasmosis. Ocular manifestations include retinitis, neuroretinitis, choroidal granulomas, peripheral granulomas and posterior pole granulomas. Infectious posterior uveitis is a challenging subject and should be considered in the differential diagnosis of any posterior uveitis in children. Infectious uveitis must be excluded before initiating immunosuppressive therapy.


Yeast as a tool to explore cathepsin D function.
Cathepsin D has garnered increased attention in recent years, mainly since it has been associated with several human pathologies. In particular, cathepsin D is often overexpressed and hypersecreted in cancer cells, implying it may constitute a therapeutic target. However, cathepsin D can have both anti- and pro-survival functions depending on its proteolytic activity, cellular context and stress stimulus. Therefore, a more detailed understanding of cathepsin D regulation and how to modulate its apoptotic functions is clearly needed. In this review, we provide an overview of the role of cathepsin D in physiological and pathological scenarios. We then focus on the opposing functions of cathepsin D in apoptosis, particularly relevant in cancer research. Emphasis is given to the role of the yeast protease Pep4p, the vacuolar counterpart of cathepsin D, in life and death. Finally, we discuss how insights from yeast cathepsin D and its role in regulated cell death can unveil novel functions of mammalian cathepsin D in apoptosis and cancer.


Epidemiology and natural history of inflammatory bowel diseases.
In the West, the incidence and prevalence of inflammatory bowel diseases has increased in the past 50 years, up to 8-14/100,000 and 120-200/100,000 persons, respectively, for ulcerative colitis (UC) and 6-15/100,000 and 50-200/100,000 persons, respectively, for Crohn's disease (CD). Studies of migrant populations and populations of developing countries demonstrated a recent, slow increase in the incidence of UC, whereas that of CD remained low, but CD incidence eventually increased to the level of UC. CD and UC are incurable; they begin in young adulthood and continue throughout life. The anatomic evolution of CD has been determined from studies of postoperative recurrence; CD begins with aphthous ulcers that develop into strictures or fistulas. Lesions usually arise in a single digestive segment; this site tends to be stable over time. Strictures and fistulas are more frequent in patients with ileal disease, whereas Crohn's colitis remains uncomplicated for many years. Among patients with CD, intestinal surgery is required for as many as 80% and a permanent stoma required in more than 10%. In patients with UC, the lesions usually remain superficial and extend proximally; colectomy is required for 10%-30% of patients. Prognosis is difficult to determine. The mortality of patients with UC is not greater than that of the population, but patients with CD have greater mortality than the population. It has been proposed that only aggressive therapeutic approaches, based on treatment of early recurrent lesions in asymptomatic individuals, have a significant impact on progression of these chronic diseases.


Effects of castration upon the morphology of the accessory sex organs of the male rat--a scanning electron microscopy study.
A systematic, comparative study of the accessory sex glands of the adult male rat after androgen withdrawal was carried out. The changes were investigated by using scanning electron microscopy at different intervals after surgical castration. The main common signs of epithelial cell involution were flattening of the cell surface, reduction of the size and number of microvilli, some blurring of the cell borders, cessation of secretory activity and diminution of the luminal volume of the glands. Overall, confident signs of atrophy were evident after one week, and complete epithelial involution was reached by the third week. The epithelial cell atrophy was accompanied by a relative stromal hyperplasia. The new observation seems to be that the process of stroma consolidation is progressing for a considerable time subsequent to the completion of the epithelial involution. This phenomenon is particularly evident in the dorsal prostate, the seminal vesicle and the coagulating gland.


Ovine congenital progressive muscular dystrophy (OCPMD) is a model of TNNT1 congenital myopathy.
Ovine congenital progressive muscular dystrophy (OCPMD) was first described in Merino sheep flocks in Queensland and Western Australia in the 1960s and 1970s. The most prominent feature of the disease is a distinctive gait with stiffness of the hind limbs that can be seen as early as 3 weeks after birth. The disease is progressive. Histopathological examination had revealed dystrophic changes specifically in type I (slow) myofibres, while electron microscopy had demonstrated abundant nemaline bodies. Therefore, it was never certain whether the disease was a dystrophy or a congenital myopathy with dystrophic features. In this study, we performed whole genome sequencing of OCPMD sheep and identified a single base deletion at the splice donor site (+ 1) of intron 13 in the type I myofibre-specific TNNT1 gene (KT218690 c.614 + 1delG). All affected sheep were homozygous for this variant. Examination of TNNT1 splicing by RT-PCR showed intron retention and premature termination, which disrupts the highly conserved 14 amino acid C-terminus. The variant did not reduce TNNT1 protein levels or affect its localization but impaired its ability to modulate muscle contraction in response to Ca2+ levels. Identification of the causative variant in TNNT1 finally clarifies that the OCPMD sheep is in fact a large animal model of TNNT1 congenital myopathy. This model could now be used for testing molecular or gene therapies.


Preinvasive Colorectal Lesions of African Americans Display an Immunosuppressive Signature Compared to Caucasian Americans.
African Americans (AAs) have higher colorectal cancer (CRC) incidence and mortality rate than Caucasian Americans (CAs). Recent studies suggest that immune responses within CRCs contribute to the disparities. If racially distinct immune signatures are present in the early phases of carcinogenesis, they could be used to develop interventions to prevent or slow disease. We selected a convenience sample of 95 patients (48 CAs, 47 AAs) with preinvasive colorectal adenomas from the surgical pathology laboratory at the Medical University of South Carolina. Using immunofluorescent-conjugated antibodies on tissue slides from the lesions, we quantified specific immune cell populations: mast cells (CD117+), Th17 cells (CD4+RORC+), and NK cell ligand (MICA/B) and inflammatory cytokines, including IL-6, IL-17A, and IFN-γ. We compared the mean density counts (MDCs) and density rate ratios (RR) and 95% CI of immune markers between AAs to CAs using negative binomial regression analysis. We adjusted our models for age, sex, clinicopathologic characteristics (histology, location, dysplasia), and batch. We observed no racial differences in age or sex at the baseline endoscopic exam. AAs compared to CAs had a higher prevalence of proximal adenomas (66% vs. 40%) and a lower prevalence of rectal adenomas (11% vs. 23%) (p =0.04) but no other differences in pathologic characteristics. In age, sex, and batch adjusted models, AAs vs. CAs had lower RRs for cells labeled with IFNγ (RR 0.50 (95% CI 0.32-0.81); p=0.004) and NK cell ligand (RR 0.67 (0.43-1.04); p=0.07). In models adjusted for age, sex, and clinicopathologic variables, AAs had reduced RRs relative to CAs for CD4 (p=0.02), NK cell ligands (p=0.01), Th17 (p=0.005), mast cells (p=0.04) and IFN-γ (p< 0.0001). Overall, the lower RRs in AAs vs. CAs suggests reduced effector response capacity and an immunosuppressive ('cold') tumor environment. Our results also highlight the importance of colonic location of adenoma in influencing these differences; the reduced immune responses in AAs relative to CAs may indicate impaired immune surveillance in early carcinogenesis. Future studies are needed to understand the role of risk factors (such as obesity) in influencing differences in immune responses by race.


Fetal hemopoiesis in diffusion chamber cultures. III. The effect of neutropenia.
Proliferation and differentiation of granulocytes, macrophages, and both myeloid committed (CFC) and pluripotent (CFU) stem cells in diffusion chamber (DC) cultures of fetal liver were studied in order to evaluate the role of circulating humoral factors in the control of fetal myelopoiesis. When DC with fetal liver cells were implanted into mice rendered neutropenic by pretreatment with cyclophosphamide, more granulocytes and CFC were produced through day 10 as compared to DC implanted into saline pretreated control hosts. A difference in CFU recovery from fetal liver suspensions grown in DC implanted into neutropenic and control hosts was not seen until day 10. Serum CSF concentrations were increased in neutropenic as compared to control host mice 2 and 3 days after implantation of DC. Levels of serum inhibitors of colony growth showed marked variability but, in general, were similar in both groups. These data provide evidence that fetal CFC and fetal myelopoiesis are influenced by a circulating humoral factor present in neutropenic serum. CSF may be the factor, although the data presented in this paper do not establish this with any certainty.


Naltrexone sustained-release/bupropion sustained-release for the management of obesity: review of the data to date.
Obesity is an emerging disease worldwide. Changes in living habits, especially with increased consumption of high-calorie foods and decreased levels of physical activity, lead to an energy imbalance that brings weight gain. Overweight and obesity are major risk factors for several chronic diseases (including cardiovascular diseases, diabetes, and cancer), reduce quality of life, and are associated with higher mortality. For all these reasons, it is of the utmost importance that the trend be reversed and obese people enabled to lose weight. It is known that eating a healthy diet and exercising regularly can help prevent obesity, but data show that in many cases these steps are not enough. This is the reason why, over the last few decades, several antiobesity drugs have been developed. However, the disappointing results demonstrated for the vast majority of them have not discouraged the pharmaceutical industry from continuing to look for an effective drug or combination of drugs. The systematic review presented here focuses on naltrexone sustained-release/bupropion sustained-release combination (Contrave(®)). We conclude from the current published reports that its effectiveness in the treatment of obesity can be estimated as a placebo-subtracted weight loss of around 4.5%. This weight reduction is moderate but similar to other antiobesity drugs. The safety profile of this combination is acceptable, despite additional data regarding cardiovascular disease being needed.


Balanced olfactory antagonism as a concept for understanding evolutionary shifts in moth sex pheromone blends.
In the sex pheromone communication systems of moths, both heterospecific sex pheromone components and individual conspecific pheromone components may act as behavioral antagonists when they are emitted at excessive rates and ratios. In such cases, the resulting blend composition does not comprise the sex pheromone of a given species. That is, unless these compounds are emitted at optimal rates and ratios with other compounds, they act as behavioral antagonists. Thus, the array of blend compositions that are attractive to males is centered around the characterized female-produced sex pheromone blend of a species. I suggest here that the resulting optimal attraction of males to a sex pheromone is the result of olfactory antagonistic balance, compared to the would-be olfactory antagonistic imbalance imparted by behaviorally active compounds when they are emitted individually or in other off-ratio blends. Such balanced olfactory antagonism might be produced in any number of ways in olfactory pathways, one of which would be mutual, gamma-aminobutyric-acid-related disinhibition by local interneurons in neighboring glomeruli that receive excitatory inputs from pheromone-stimulated olfactory receptor neurons. Such mutual disinhibition would facilitate greater excitatory transmission to higher centers by projection interneurons arborizing in those glomeruli. I propose that in studies of moth sex pheromone olfaction, we should no longer artificially compartmentalize the olfactory effects of heterospecific behavioral antagonists into a special category distinct from olfaction involving conspecific sex pheromone components. Indeed, continuing to impose such a delineation among these compounds may retard advances in understanding how moth olfactory systems can evolve to allow males to exhibit correct behavioral responses (that is, attraction) to novel sex-pheromone-related compositions emitted by females.


Expansion of CD25-Negative Forkhead Box P3-Positive T Cells during HIV and Mycobacterium tuberculosis Infection.
Tuberculosis (TB) and HIV alter the immune system, and coinfected (HIV-TB) individuals usually present deregulations of T-lymphocytic immune response. We previously observed an increased frequency of "unconventional" CD4+CD25-FoxP3+ Treg (uTreg) population during HIV-TB disease. Therefore, we aimed to explore the phenotype and function of uTreg and conventional CD4+CD25+FoxP3+ Treg subsets (cTreg) in this context. We evaluated the expression of CD39, programmed cell death protein 1 (PD1), glucocorticoid-induced tumor necrosis factor receptor (GITR), and the effector/memory distribution by flow cytometry in cTreg and uTreg. Also, IL-10, TGF-β, IFN-γ production, and the suppressor capacity of uTregs were analyzed in cocultures with effector lymphocytes and compared with the effect of regulatory T cells (Tregs). We found diminished expression of CD39 and higher levels of PD1 on uTreg compared to cTreg in both HIV-TB and healthy donors (HD). In addition, uTreg and cTreg showed differences in maturation status in both HIV-TB and HD groups, due to the expansion of effector memory uTregs. Interestingly, both HIV-TB and HD showed a pronounced production of IFN-γ in uTreg population, though no significant differences were observed for IL-10 and TGF-β production between uTreg and cTreg. Moreover, IFN-γ+ cells were restricted to the CD39- uTreg population. Finally, when the suppressor capacity was evaluated, both uTreg and cTreg inhibited polyclonal T cell-proliferation and IFN-γ production in a similar extent. These findings suggest that uTregs, which are expanded during HIV-TB coinfection, exert regulatory functions in a similar way to cTregs despite an altered surface expression of Treg characteristic markers and differences in cytokine production.


Ventilator management of infants before extracorporeal membrane oxygenation.
The aim of this project was to review the course of infants referred for consideration of extracorporeal membrane oxygenation (ECMO) to identify maximal ventilator settings that, when exceeded, did not provide clinical benefit to the patient. These settings might then be used in defining failure of conventional mechanical ventilation. We reviewed referral records and hospital charts of all infants treated for severe respiratory failure due to meconium aspiration syndrome during the 52.5 month period from March 15, 1985, to August 1, 1989. At an inspiratory pressure > 35 cm H2O, 75% (43/57) of patients eventually required ECMO, and 28% (4/14) of the infants who did not receive ECMO died. When the inspiratory pressure was > or = 40 cm H2O, 39/49 patients required ECMO, and 30% (3/10) of those not treated with ECMO died. Once the inspiratory pressure was > 45 cm H2O, 91% (29/32) of patients required ECMO, and only one third of those not treated with ECMO survived. Although the limitations for conventional therapy suggested in this paper may be helpful to clinicians, each center needs to establish guidelines for maximal conventional ventilator support. If these guidelines are clearly defined, alternative methods of therapy can be used once these criteria are achieved.


Causal superlearning arising from interactions among cues.
Superconditioning refers to supernormal responding to a conditioned stimulus (CS) that sometimes occurs in classical conditioning when the CS is paired with an unconditioned stimulus (US) in the presence of a conditioned inhibitor for that US. In the present research, we conducted 4 experiments to investigate causal superlearning, a phenomenon in human causal learning analogous to superconditioning. Experiment 1 demonstrated superlearning relative to appropriate control conditions. Experiment 2 showed that superlearning wanes when the number of cues used in an experiment is relatively large. Experiment 3 determined that even when relatively many cues are used, superlearning can be observed provided testing is conducted immediately after training, which is problematic for explanations by most contemporary learning theories. Experiment 4 found that ratings of a superlearning cue are weaker than those to the training excitor which gives basis to the conditioned inhibitor-like causal preventor used during causal superlearning training. This is inconsistent with the prediction by propositional reasoning accounts of causal cue competition, but is readily explained by associative learning models. In sum, the current experiments revealed some weaknesses of both the associative and propositional reasoning models with respect to causal superlearning. (PsycINFO Database Record


Experiences of mothers of infants with neonatal abstinence syndrome in the neonatal intensive care unit.
To describe the experiences of mothers of infants with neonatal abstinence syndrome (NAS) in the neonatal intensive care unit (NICU). Qualitative description. We recruited participants from community-based, out-patient, addiction treatment facilities in a large urban city in the southwestern region of the United States. A convenience sample of 15 Hispanic, substance addicted mothers of infants with NAS participated. We conducted semistructured, individual, interviews and analyzed the data using qualitative content analysis. First, we analyzed the data independently and then discussed the themes until a consensus was reached. We identified four themes: (a) understanding addiction, (b) watching the infant withdraw, (c) judging, and (d) trusting the nurses. The participants felt there was a lack of understanding concerning addiction that was particularly noted when interacting with the nurses. They shared their feelings of guilt and shame when observing their infants withdrawing. The participants felt judged by the nurses for having used illicit drugs during pregnancy. Feeling judged interfered with the participants' ability to trust the nurses. These findings provide nurses with a better understanding of the experiences of mothers who have addiction problems and may lead to more customized nursing care for this high-risk population of mothers and their infants.


Eicosapentaenoic Acid and Vitamin E Against Doxorubicin Induced Cardiac and Renal Damages: Role of Cytochrome c and iNOS.
The current study aimed to evaluate the mechanisms involved in protection against doxorubicin-induced cardiac and renal toxicities upon treatment with eicosapentaenoic acid and vitamin E. Rats were randomly assigned to 4 groups: normal control, doxorubicin inducted control, eicosapentaenoic acid treated group and a final group pretreated with vitamin E. Lipid peroxidation, reduced glutathione (GSH) and tumor necrosis factor-alpha (TNF-α) contents as well as glutathione peroxidase (GPx), superoxide dismutase (SOD) and myeloperoxidase (MPO) activities were assessed. Moreover, hearts were used for immunohistochemical detection of the pro-apoptotic protein cytochrome c expression, while the kidneys were used for detection of inducible nitric oxide synthase (iNOS) expression. Eicosapentaenoic acid and vitamin E produced significant protection from doxorubicin-induced cardiac and renal toxicities. The suggested mechanisms for protection included decreased cytochrome c and iNOS expression as well as markedly decreased lipid peroxides and TNF-α contents accompanied with increased GSH content as compared to the doxorubicin control group. Moreover, there was marked increase in GPx and SOD activities accompanied by significant suppression of MPO activity. The present study demonstrated the potent protective effects of eicosapentaenoic acid and vitamin E from doxorubicin induced cardiac and renal toxicities through their potent anti-oxidant, anti-inflammatory and anti-apoptotic properties. Hence, eicosapentaenoic acid and vitamin E could be promising protective agents against doxorubicintoxicity.


Meat consumption and diet quality and mortality in NHANES III.
There is growing evidence that meat consumption is associated with total and cause-specific mortality. Our objective was to evaluate the association of meat intake and the healthy eating index (HEI) with total mortality, cancer and cardiovascular disease (CVD) mortality. Analyses are based on 17, 611 participants from Third National Health and Nutrition Examination Survey (NHANES III) (1986-2010). Meat intake was assessed using a food frequency questionnaire administrated at baseline. Adherence to the HEI was analyzed with a single 24-h dietary recall. Cox proportional hazards regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) of mortality according to five categories of meat consumption and three categories of the HEI score. During the follow-up period, 3683 deaths occurred, of which 1554 were due to CVD and 794 due to cancer. After multivariable adjustment, neither red and processed meat, nor white meat consumption were consistently associated with all-cause or cause-specific mortality. In men, white meat consumption tended to be inversely associated with total mortality (P for trend=0.02), but there was no such association among women. Significantly decreased mortality was observed in the top compared with the bottom third of the HEI score (HR=0.70, 95% CI 0.52-0.96). This association was only observed in men, but not in women. Meat consumption was not associated with mortality. A healthy diet according to HEI, however, was associated with a decreased total mortality in men, but not in women.


Placental lactogen levels as guide to outcome of threatened abortion.
The clinical value has been assessed of circulating placental lactogen levels as a pointer to the outcome in a patient with vaginal bleeding in early pregnancy. By using a semiautomated radioimmunoassay the normal range of values for the first and second trimesters has been established. In patients admitted with vaginal bleeding after the eighth week of gestation estimation of plasma human placental lactogen showed that patients with low levels were those in whom the abortion was completed during the first admission. Women whose pregnancies continued normally or who aborted after their first discharge from hospital had normal levels. In a small group sampled before the onset of bleeding but who later aborted the mean levels were lower than normal. This simple and inexpensive test can indicate those women in whom abortion is inevitable and could be used to reduce substantially the length of hospital stay in this common complication of early pregnancy.


COVID-19 pneumonia treated with ultra-low doses of radiotherapy (ULTRA-COVID study): a single institution report of two cases.
Since the outbreak of coronavirus disease 2019 (COVID-19) pandemic, healthcare systems have focused their efforts into finding a treatment to avoid the fatal outcomes of severe acute respiratory syndrome due to coronavirus‑2 (SARS-CoV-2). Benefits and risks of systemic treatments remain unclear, with multiple clinical trials still ongoing. Radiotherapy could play a role in reducing the inflammatory response in the lungs and relieve life-threatening symptoms. We designed a prospective study of Ultra-Low Doses of Therapy with Radiation Applied to COVID-19 (ULTRA-COVID) for patients who suffer pneumonia, are not candidates for invasive mechanical ventilation and show no improvement with medical therapy. We present the preliminary results of two patients diagnosed with COVID-19 pneumonia treated with ULTRA-COVID. After one radiotherapy session, significant clinical response and a good radiological response was observed in both cases, resulting in both patients being discharged from hospital in less than 2 weeks after radiation treatment. Preliminary clinical and radiological results suggest a potential benefit of treating COVID-19 pneumonia with ULTRA-COVID. ClinicalTrials.gov Identifier: NCT04394182.


A Mindfulness-Based Intervention to Alleviate Stress From Discrimination Among Young Sexual and Gender Minorities of Color: Protocol for a Pilot Optimization Trial.
Young sexual and gender minorities (SGMs) of color may face unique experiences of discrimination based on their intersectional positions (eg, discrimination based on both racial or ethnic identity and sexual identity). Emerging evidence suggests that mindfulness practices may reduce stress from discrimination and improve overall well-being among young SGM. Moreover, the omnipresence of smartphone access among racial or ethnic and sexual minority communities provides a method through which to administer mindfulness-based interventions among young SGMs of color. This paper outlines the protocol of the Optimizing a Daily Mindfulness Intervention to Reduce Stress from Discrimination among Young Sexual and Gender Minorities of Color (REDUCE) study, a pilot optimization trial of a smartphone-based mindfulness intervention that was developed in conjunction with the Healthy Minds Program (HMP) with the aim of reducing stress from discrimination among young SGMs. In total, 80 young (ages 18-29 years) SGMs of color will be enrolled in the study. The HMP is a self-guided meditation practice, and participants will be randomized to either a control condition or an intervention that uses a neuroscience-based approach to mindfulness. We will use the multiphase optimization strategy to assess which combination of mindfulness interventions is the most effective at reducing stress from discrimination among young SGMs of color. A combination of mindfulness-based meditation intervention components will be examined, comprising mindfulness-based practices of awareness, connection, and purpose. Awareness refers to the practice of self-awareness, which reduces the mind's ability to be distracted and instead be present in the moment. Connection refers to the practice of connection with oneself and others and emphasizes on empathy and compassion with oneself and others. Purpose encourages goal-making in accordance with one's values and management of behavior in accordance with these goals. In addition, we will assess the feasibility and acceptability of the HMP application among young SGMs of color. The REDUCE study was approved by the Institutional Review Board of New York University, and recruitment and enrollment began in the winter of 2021. We expect to complete enrollment by the summer of 2022. The results will be disseminated via social media, journal articles, abstracts, or presentations, as well as to participants, who will be given the opportunity to provide feedback to the researchers. This optimization trial is designed to test the efficacy, feasibility, and acceptability of implementing an application-based, mindfulness-based intervention to reduce stress from discrimination and improve well-being among young SGMs of color. Evidence from this study will assist in the creation of a sustainable, culturally relevant mobile app-based mindfulness intervention to reduce stress from discrimination among young SGMs of color. Clinicaltrials.gov NCT05131360; https://clinicaltrials.gov/ct2/show/NCT05131360. DERR1-10.2196/35593.


Use of intravenous immunoglobulin in pediatric practice.
In recent years, human-driven intravenous immunoglobulins (IVIG) administered intravenously have been widely used in treatment of many diseases. Intravenous immunoglobulin is obtained from human-driven plasma pools as in other plasma-driven products and IVIG preperations contain structurally and functionally intact immunoglobulin. Intravenous immunoglobulin was approved by FDA (Food and Drug Administration) in USA in 1981 for the first time and was started to be primarily used in patients with immune deficiency with hypogammaglobulinemia. The effects of intravenous immunoglobulin include complex mechanisms, but it exerts its essential action by eliminating the non-specific Fc receptors found in the mononuclear phagocytic system or by inhibiting binding of immune complexes to Fc receptors in the cells. Their areas of usage include conditions where their anti-inflammatory and immunomudulator effects are utilized in addition to replacement of deficient immunoglobulin. Although the definite indications are limited, it has been shown that it is useful in many diseases in clinical practice. Its side effects include fever, sweating, nausea, tachycardia, eczematous reactions, aseptic meningitis, renal failure and hematological-thromboembolic events. In this article, use of IVIG, its mechanisms of action, indications and side effects were discussed.


Elevated ATG13 in serum of patients with ME/CFS stimulates oxidative stress response in microglial cells via activation of receptor for advanced glycation end products (RAGE).
Myalgic Encephalomyelitis, also known as Chronic Fatigue Syndrome (ME/CFS), is a multisystem illness characterized by extreme muscle fatigue associated with pain, neurocognitive impairment, and chronic inflammation. Despite intense investigation, the molecular mechanism of this disease is still unknown. Here we demonstrate that autophagy-related protein ATG13 is strongly upregulated in the serum of ME/CFS patients, indicative of impairment in the metabolic events of autophagy. A Thioflavin T-based protein aggregation assay, array screening for autophagy-related factors, densitometric analyses, and confirmation with ELISA revealed that the level of ATG13 was strongly elevated in serum samples of ME/CFS patients compared to age-matched controls. Moreover, our microglia-based oxidative stress response experiments indicated that serum samples of ME/CFS patients evoke the production of reactive oxygen species (ROS) and nitric oxide in human HMC3 microglial cells, whereas neutralization of ATG13 strongly diminishes the production of ROS and NO, suggesting that ATG13 plays a role in the observed stress response in microglial cells. Finally, an in vitro ligand binding assay provided evidence that ATG13 employs the Receptor for Advanced Glycation End-products (RAGE) to stimulate ROS in microglial cells. Collectively, our results suggest that an impairment of autophagy following the release of ATG13 into serum could be a pathological signal in ME/CFS.


Effects of Suvorexant, a Dual Orexin/Hypocretin Receptor Antagonist, on Impulsive Behavior Associated with Cocaine.
Hypothalamic hypocretin (orexin) peptides mediate arousal, attention, and reward processing. Fibers containing orexins project to brain structures that govern motivated behavior, including the ventral tegmental area (VTA). A number of psychiatric conditions, including attention deficit hyperactivity disorder (ADHD) and substance use disorders, are characterized by deficits in impulse control, however the relationship between orexin and impulsive behavior is incompletely characterized. The effects of systemic or centrally administered orexin receptor (OXR) antagonists on measures of impulsive-like behavior in rats were evaluated using the five-choice serial reaction time task (5-CSRTT) and delay discounting procedures. These paradigms were also used to test the capacity of OXR antagonists to attenuate acute cocaine-evoked impulsivity. Finally, immunohistochemistry and calcium imaging were used to assess potential cellular mechanisms by which OXR blockade may influence motor impulsivity. Suvorexant, a dual (OX1/2R) orexin receptor antagonist, reduced cocaine-evoked premature responses in 5-CSRTT when administered systemically or directly into VTA. Neither suvorexant nor OX1R- or OX2R-selective compounds (SB334867 or TCS-OX2-29, respectively) altered delay discounting. Finally, suvorexant did not alter Fos-immunoreactivity within tyrosine hydroxylase-immunolabeled neurons of VTA, but did attenuate cocaine- and orexin-induced increases in calcium transient amplitude within neurons of VTA. Results from the present studies suggest potential therapeutic utility of OXR antagonists in reducing psychostimulant-induced motor impulsivity. These findings also support the view that orexin transmission is closely involved in executive function in normal and pathological conditions.


Chitosan scaffolds with a shape memory effect induced by hydration.
We demonstrate that chitosan-based porous scaffolds can present a shape memory effect triggered by hydration. The shape memory effect of non-crosslinked (CHT0) and genipin-crosslinked (CHT1) scaffolds was followed by innovative hydromechanical compressive tests and dynamic mechanical analysis (DMA), while the sample was immersed in varying compositions of water-ethanol mixtures. By dehydration with higher contents of ethanol, the vitreous-like nature of the amorphous component of chitosan allows the fixation of the temporary shape of the scaffold. The presence of water disrupts inter-molecular hydrogen bonds permitting large-scale segmental mobility of the chitosan chains upon the occurrence of glass transition and thus the recovery of the permanent shape of a pre-deformed scaffold. Results showed that chitosan possesses shape memory properties, characterized by a fixity ratio above 97.2% for CHT0 and above 99.2% for CHT1 and a recovery ratio above 70.5% for CHT0 and 98.5% for CHT1. In vitro drug delivery studies were also performed to demonstrate that such devices can also be loaded with molecules. We show that the developed chitosan scaffolds are candidates of biomaterials for applications in minimally invasive surgery for tissue regeneration or for drug delivery.


Thai people living with tuberculosis and how they adhere to treatment: A grounded theory study.
To develop a conceptual framework of adherence to treatment among Thai people living with tuberculosis, a grounded theory approach was used. A purposive sample of 20 Thai people living with tuberculosis, aged from 23 to 85 years, was interviewed. From the participants' perspective, a core category of social belonging was highlighted, with three categories of conditions connected: personal barriers, personal resilience, and social facilitation. Personal barriers encompassed fear of stigma, concealing the illness, and lack of knowledge and motivation to complete the treatment regime. Personal resilience encompassed positive thinking and self-awareness. Social facilitation encompassed the ease of access to health services, continuity in the health service's ability to choose a directly-observed therapy observer, and social support. This study contributes a deeper understanding of the perspective of Thai people living with tuberculosis with regards to adherence to tuberculosis treatment. It might improve how local healthcare workers provide tuberculosis care, and inspire them to tailor care to people living with tuberculosis in a local community to increase personal resilience and reduce stigma.


Inactivation of the cytochrome P450 gene CYP82E2 by degenerative mutations was a key event in the evolution of the alkaloid profile of modern tobacco.
The alkaloid profile of cultivated tobacco (Nicotiana tabacum) is different from that of its two progenitors, Nicotiana sylvestris and Nicotiana tomentosiformis, in that tobacco accumulates nicotine as the most abundant alkaloid, while its ancestors convert nicotine to nornicotine in the senescing leaf. The nicotine-retaining phenotype of tobacco is thought to have evolved through the inactivation of the conversion loci inherited from its two progenitors. Here, the genetic changes associated with the inactivation of the conversion locus derived from N. sylvestris were investigated. Candidate genes were isolated from a N. sylvestris senescing leaf cDNA library and characterized by heterologous gene expression in yeast, site-directed mutagenesis and quantitative real-time polymerase chain reaction. A cytochrome P450 gene, designated NsylCYP82E2, was isolated from N. sylvestris. Located on the chromosomal fragment defined by the N. sylvestris conversion locus, NsylCYP82E2 confers high nicotine N-demethylase (NND) activity in the senescing leaves of N. sylvestris, but the gene is inactivated by two degenerative mutations in tobacco. Collectively with previously published data, these results show that inactivation of NND genes by degenerative mutations and/or transcriptional suppression played a key role in the evolution of the alkaloid profile of modern tobacco.


Altered fluxes responsible for reduced hepatic glucose production and gluconeogenesis by exogenous glucose in rats.
The net release of glucose from the liver, or hepatic glucose production (HGP), and apparent gluconeogenesis (GNG) are reduced by exogenous glucose. We investigated the changes in metabolic fluxes responsible. Flux through the hepatic GNG pathway was quantified by mass isotopomer distribution analysis (MIDA) from [2-13C]glycerol. Unidirectional flux across hepatic glucose-6-phosphatase (G-6-Pase), or total hepatic glucose output (THGO), and hepatic glucose cycling (HGC) were also measured by using glucuronate (GlcUA) to correct for glucose 6-phosphate (G-6-P) labeling. Infusion of glucose (15-30 mg.kg-1.min-1 iv) to 24 h-fasted rats caused two important metabolic alterations. First was a significant increase in hepatic glucose uptake and HGC: > 60% of THGO was from HGC. Second, although flux through hepatic G-6-P increased (from 15.7 to 17.7-22.7 mg.kg-1.min-1), the partitioning of G-6-P flux changed markedly [from 30-35% to 55-60% entering UDP-glucose (UDP-Glc), P < 0.01]. Total flux through the GNG pathway remained active during intravenous glucose, but increased partitioning into UDP-Glc lowered GNG flux plasma glucose by 50%. In summary, the suppression of HGP and GNG flux into glucose is not primarily due to reduced carbon flow through hepatic G-6-Pase or the hepatic GNG pathway. THGO persists, but hepatic G-6-P is derived increasingly from plasma glucose, and flow through GNG persists, but the partitioning coefficient of G-6-P into UDP-Glc doubles. These adjustments permit net HGP to fall despite increased total production of hepatic G-6-P during administration of glucose.


[Retrospective evaluation of various methods of examination for prognosis of cardiac insufficiency in the subacute period of myocardial infarct].
A retrospective analysis of the informativeness of certain methods of examination for ascertaining the possibility of cardiac failure developing in the subacute period of penetrating, predominantly primary transmural myocardial infarction with no clinical signs of cardiac failure in the first three days after the onset of the disease. In 60 patients circulation time in the "arm--ear" area and in 25 of them the stroke and minute volumes by the dye dilution method were studied. The state of the fluid spaces was investigated in 18 patients and daily natriuresis in 23. In 36 patients the condition of the pulmonary circulation was judged by means of roentgenogram. The high value of the information gained from study of circulation time and natriuresis and X-ray examination in the prognostication of the development of cardiac failure is shown.


Spatial and temporal variability of fecal indicator bacteria in an urban stream under different meteorological regimes.
As a representative urban stream in Korea, the Gwangju (GJ) stream suffers from chronic fecal contamination. In this study, to characterize levels of fecal pollution in the GJ stream, the monthly monitoring data for seven years (from 2001 to 2007) and the hourly monitoring data from two field experiments were examined with respect to seasonal/daily variations and spatial distribution under wet and dry weather conditions. This research revealed that concentrations of fecal indicator bacteria strongly varied depending on the prevalent meteorological conditions. That is, during the dry daytime, fecal indicator bacteria concentrations decreased due to inactivation from solar irradiation, but rapidly increased in the absence of sunlight, suggesting external source inputs. In addition, bacterial concentrations substantially increased during rainfall events, due probably to a major contribution from combined sewer overflow. The observations in this study can be useful for implementing fecal pollution management strategies and for predicting fecal contamination as a function of meteorological conditions.


Assessment of the effects of gabapentin on activity levels and owner-perceived mobility impairment and quality of life in osteoarthritic geriatric cats.
OBJECTIVE Toevaluate effects of gabapentin on activity levels and owner-perceived mobility impairment and quality of life (QOL) in osteoarthritic geriatric cats. DESIGN Blinded, placebo-controlled, randomized crossover-design study. ANIMALS 20 osteoarthritic cats (≥ 10 years old). PROCEDURES Cats received gabapentin (10 mg/kg [4.5 mg/lb]) or placebo treatment, PO, every 12 hours for 2 weeks, followed by the alternate treatment (with no washout period). Activity was assessed with a collar-mounted accelerometer. A client-specific outcome measure (CSOM) questionnaire was used weekly to collect owner assessments of 3 selected activities in which their cats had impaired mobility; QOL ratings (worse, the same, or improved) following crossover to each treatment and for the overall study period were collected at the end of the investigation. Activity counts, CSOM and QOL data, and deterioration in impaired activities (ie, decrease of ≥ 2 points in CSOM scores) associated with treatment crossover were assessed statistically. Adverse events were recorded. RESULTS Gabapentin administration was associated with significantly lower mean daily activity counts (48,333 vs 39,038 counts/d) and significantly greater odds (approx 3-fold change) of CSOM ratings indicating improvement in impaired activities, compared with results for the placebo treatment. A greater proportion of cats had deterioration in impaired activities after the crossover from gabapentin to placebo than when the opposite occurred, but the proportion of cats with worsened QOL did not differ between sequences. Adverse events were noted for 10 cats (9 that completed the study) during gabapentin treatment (sedation, ataxia, weakness, and muscle tremors) and 1 cat during placebo treatment (lethargy). CONCLUSIONS AND CLINICAL RELEVANCE Gabapentin treatment was associated with improvement in owner-identified impaired activities of osteoarthritic cats. Activity levels were lower than those during placebo treatment, and sedation was the most common adverse effect.


[Non traumatic lesions of the foot, calluses and nails: socioeconomic impact of an unexplored issue].
Up to 20% of the general population, especially women and old people, suffer from non traumatic lesions of the foot. These lesions have repercussions on a person's health and wellbeing. Nevertheless, the clinical examination of the foot rarely forms part of routine medical consultations. Using questionnaires and individual interviews, this pilot study investigated the foot problems of 96 patients attending a podiatrist's practice, as well as the level of communication between the patients and their doctor. 85% of the participants said they never talked about calluses and nails with their doctor. The results were confirmed by fifteen individual interviews. Clinicians have an important role to play in the early detection of these lesions, allowing an efficient management of this issue.


Reliability and clinically important improvement thresholds for osteoarthritis pain and function scales: a multicenter study.
To assess the reliability and clinically meaningful thresholds of intermittent and constant osteoarthritis pain (ICOAP) score, the Knee injury and Osteoarthritis Outcome Score Physical function Short-form (KOOS-PS), the Hip disability and Osteoarthritis Outcome Score Physical function Short-form (HOOS-PS), and the Quality of life subscales of HOOS/KOOS (HOOS-QOL/KOOS-QOL) in patients with knee or hip arthritis. One hundred and ninety-five patients (141 knee, 54 hip) seen at 2 orthopedic outpatient clinics with a diagnosis of knee or hip OA completed patient-reported questionnaires (ICOAP pain scale, KOOS-PS, HOOS-PS, KOOS-QOL, HOOS-QOL) at baseline and 2-week followup. Reliability was assessed using intraclass correlation coefficients (ICC). We calculated minimum clinically important difference (MCID) and moderate improvement in the subgroup that reported change in the status of their affected joint. The reliability as assessed by ICC was as follows: ICOAP pain scale, 0.63 (0.48, 0.74) in patients with knee arthritis, and 0.86 (0.73, 0.93) for hip arthritis; KOOS-PS, 0.66 (0.52, 0.77); HOOS-PS, 0.82 (0.66, 0.91); KOOS-QOL, 0.79 (0.69, 0.86); and HOOS-QOL, 0.67 (0.42, 0.83). MCID and moderate improvement estimates in patients with knee arthritis were ICOAP pain scale, 18.5 and 26.7; KOOS-PS, 2.2 and 15.0; and KOOS-QOL, 8.0 and 15.6. A smaller sample in patients with hip arthritis precluded MCID and moderate improvement estimates. We found that ICOAP pain and KOOS-PS/HOOS-PS scales were reasonably reliable in patients with hip OA. Reliability of these scales was much lower in patients with knee arthritis. Thresholds for clinically meaningful change in pain or function on these scales were estimated for patients with knee arthritis.


Patient satisfaction with spinal cord stimulation for predominant complaints of chronic, intractable low back pain.
Results of subsequent surgical intervention in patients with intractable pain after lumbar spine surgery are typically worse than for initial surgery, particularly in those with predominant complaints of back pain rather than lower extremity pain. Spinal cord stimulation (SCS) has been found to yield good results in patients with primary complaints of intractable lower extremity pain. Technological advances have broadened the indications for this treatment. The purpose of this study was to evaluate patient satisfaction after SCS in the treatment of patients with predominant complaints of chronic, intractable, low back pain. Data were collected from retrospective chart review and patient follow-up questionnaire. Patients were treated at a spine specialty center. The study group consisted of the consecutive series of our first 41 patients who underwent SCS for predominant complaints of low back pain. The mean symptom duration was 82.9 months, and the mean age was 47.9 years (range, 28-83 years). All but three patients had previously undergone lumbar spine surgery (mean, 2.3 prior surgeries). At the time of follow-up (5.5-19 months after SCS implantation), patients completed questionnaires assessing their satisfaction with their outcome, if they would have the procedure again knowing what their outcome would be and if they would recommend SCS to someone with similar problems. In determining outcome, a negative response was assigned for patients who had the device removed. A worst-case analysis was also conducted in which a negative response was assigned for patients lost to follow-up or who failed to respond to a particular question. Data were also collected on complications and re-operations. All trial stimulation procedures were performed under local anesthetic with the patient providing feedback concerning pain relief achieved with various lead placements and settings. If one lead did not provide acceptable relief in all the areas needed, placement of a second lead was pursued. If the patient failed to maintain acceptable pain relief (> or =50% pain relief) during a multiday trial period, the leads were removed. If adequate relief was maintained during the trial period, the receiver was implanted. Responses to the follow-up questionnaire indicated that 60% of patients considered themselves improved from their preoperative condition and the remaining 40% did not; 78.1% of patients would recommend SCS to someone with similar problems, 69.0% were satisfied, 75.0% would have the procedure performed again if they had known their outcome before implantation. Among the 36 patients in whom the system was implanted, it was later removed in 4 because of lack of sufficient pain relief. Other re-operations included repositioning of the leads to regain pain relief in the areas needed, replacement of a malfunctioning unit and revision of lead extension wires. In this retrospective study, the majority of patients were satisfied with the results of SCS and would have the procedure again knowing what their outcome would be. These results suggest that further investigation of SCS is warranted in this difficult to treat patient population presenting with predominant complaints of chronic, intractable, axial low back pain.


Forward-time simulation of realistic samples for genome-wide association studies.
Forward-time simulations have unique advantages in power and flexibility for the simulation of genetic samples of complex human diseases because they can closely mimic the evolution of human populations carrying these diseases. However, a number of methodological and computational constraints have prevented the power of this simulation method from being fully explored in existing forward-time simulation methods. Using a general-purpose forward-time population genetics simulation environment, we developed a forward-time simulation method that can be used to simulate realistic samples for genome-wide association studies. We examined the properties of this simulation method by comparing simulated samples with real data and demonstrated its wide applicability using four examples, including a simulation of case-control samples with a disease caused by multiple interacting genetic and environmental factors, a simulation of trio families affected by a disease-predisposing allele that had been subjected to either slow or rapid selective sweep, and a simulation of a structured population resulting from recent population admixture. Our algorithm simulates populations that closely resemble the complex structure of the human genome, while allows the introduction of signals of natural selection. Because of its flexibility to generate different types of samples with arbitrary disease or quantitative trait models, this simulation method can simulate realistic samples to evaluate the performance of a wide variety of statistical gene mapping methods for genome-wide association studies.


[Executive Summary of ARIA 2019: Integrated care pathways for allergic rhinitis in Argentina, Spain and Mexico].
The health and economic impact of allergic diseases are increasing rapidly, and changes in management strategies are required. Its influence reduces the capacity of work and school performance by at least a third. The ICPs of the airways (integrated care pathways for respiratory diseases) are structured multidisciplinary healthcare plans, promoting the recommendations of the guidelines in local protocols and their application to clinical practice. This document presents an executive summary for Argentina, Mexico, and Spain. Next-generation ARIA guidelines are being developed for the pharmacological treatment of allergic rhinitis (AR), using the GRADE-based guidelines for AR, tested with real-life evidence provided by mobile technology with visual analogue scales. It is concluded that in the AR treatment, H1-antihistamines are less effective than intranasal corticosteroids (INCS), in severe AR the INCS represent the first line of treatment, and intranasal combination INCS + anti-H1 is more effective than monotherapy. However, according to the MASK real-life observational study, patients have poor adherence to treatment and often self-medicate, according to their needs.


Reproductive and teratogenic effects of nitrous oxide, isoflurane, and their combination in Sprague-Dawley rats.
The reproductive and teratogenic effects of nitrous oxide (N2O), isoflurane, and their combination were studied in 130 timed-pregnant rats. Rats were exposed to either air, 0.35% isoflurane (1/4 MAC), 50% N2O (a known teratogenic concentration), or 50% N2O plus 0.35% isoflurane for 24 h on day 8 of pregnancy. On day 20 of pregnancy, cesarean sections were performed; a total of 1268 offspring were delivered and immediately examined for external abnormalities. They were subsequently examined microscopically either for visceral or skeletal abnormalities. N2O caused significantly higher incidences of early and late resorptions, and major visceral malformations. The addition of isoflurane to N2O prevented the majority of these adverse effects. These results cast doubt on the methionine synthase inhibition theory of N2O teratogenicity.


V79 Chinese-hamster cells rendered resistant to high cadmium concentration also become resistant to oxidative stress.
Chinese hamster cells (V79) resistant to high concentrations of Cd2+ in the medium were obtained by using the procedure of Beach & Palmiter [(1981) Proc. Natl. Acad. Sci. U.S.A. 78, 2110-2114], which in mouse led to amplification of metallothionein (MT) genes and to an enrichment in cellular MT. The Cd-resistant V79 clones isolated were significantly more resistant than parental cells to oxidative stress by extracellular H2O2 or a mixture of H2O2 and superoxide anion (O2-) generated by xanthine oxidase plus acetaldehyde. On a per-cell basis, there was no difference between the two cells in their total H2O2-decomposing or O2-(-)dismutating activity. The most likely explanation is that an enrichment in MT content in the Cd-resistant cells was responsible for this effect, because of the antioxidant properties already described for this protein.


Plasmodium falciparum merozoite surface protein 1 blocks the proinflammatory protein S100P.
The malaria parasite, Plasmodium falciparum, and the human immune system have coevolved to ensure that the parasite is not eliminated and reinfection is not resisted. This relationship is likely mediated through a myriad of host-parasite interactions, although surprisingly few such interactions have been identified. Here we show that the 33-kDa fragment of P. falciparum merozoite surface protein 1 (MSP1(33)), an abundant protein that is shed during red blood cell invasion, binds to the proinflammatory protein, S100P. MSP1(33) blocks S100P-induced NFκB activation in monocytes and chemotaxis in neutrophils. Remarkably, S100P binds to both dimorphic alleles of MSP1, estimated to have diverged >27 Mya, suggesting an ancient, conserved relationship between these parasite and host proteins that may serve to attenuate potentially damaging inflammatory responses.


Dihydrokaempferol attenuates LPS-induced inflammation and apoptosis in WI-38 cells.
Globally, pneumonia has been associated as a primary cause of mortality in children aged less than 5 years. Dihydrokaempferol (DHK) has been proposed for being correlated with the process of various diseases. Nevertheless, whether DHK has a role in the progression of infantile pneumonia remains unclear. This study aimed at exploring whether DHK was involved in the progression of infantile pneumonia. Human fibroblast cells WI-38 were treated with lipopolysaccharide (LPS). The viability of WI-38 cells was measured via Cell counting kit-8. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed the protein levels of IL-1β, IL-6, TNF-α, Bax, and cleaved-caspase 3. Flow cytometry was applied for exploring the apoptosis of WI-38 cells. The concentrations of IL-1β, IL-6, and TNF-α were assessed via enzyme-linked-immunosorbent serologic assay. DHK modulated the viability of WI-38 cells in infantile pneumonia. Furthermore, we identified that DHK treatment inversely changed LPS induction-mediated elevation on the levels of inflammation biomarkers. Besides, DHK counteracted LPS-induced production of reactive oxygen species (ROS) in WI-38 cells. DHK also decreased LPS-induced elevation of WI-38 cells apoptosis and mediated the levels of apoptosis-associated indexes. Moreover, modulating sirtuin-1 (SIRT1) protein level was lowered by the induction of LPS, and was reversed by DHK treatment. In addition, DHK counteracted LPS induction-mediated elevation of p-p65 and phosphorylated inhibitor of nuclear factor kappa-B kinase subunit alpha (p-IκBα) protein levels. DHK alleviated LPS-induced WI-38 cells inflammation injury in infantile pneumonia through SIRT1/NF-κB pathway. The results shed light on the implications of DHK on the prevention and treatment of infantile pneumonia.


Early decrease in intermediate monocytes in peripheral blood is characteristic of multiple system atrophy-cerebellar type.
To identify biomarkers for multiple system atrophy-cerebellar type (MSA-C), we used flow cytometry to measure surface marker expression of peripheral blood monocytes from patients with MSA-C or hereditary spinocerebellar degeneration (hSCD) and from healthy controls (HCs). The percentage of intermediate monocytes was significantly lower in MSA-C patients than in hSCD patients and HCs and showed significant positive correlations with disease duration and unified MSA rating scale scores. The percentage of CD62L+ intermediate monocytes was significantly lower in MSA-C patients than in hSCD patients and HCs. Early decrease of peripheral blood intermediate monocytes is characteristic of MSA-C and is a biomarker.


Brain cells in culture: morphological transformation by a protein.
One type of elmbryonic rat brain cell having an epithelioid morphology in the monolayer culture can be transformed by brain extract into cells having extensive processes resembling mature astrocytes. The transforming factor is a protein with a molecular weight of 350,000. A partially purified sample showed that it is active at a concentration as low as 1 x 10(-8)M. The transforming actvity is high in adult brains but low in embryonic brains and tumors of the nervous systems.


Striatal and prefrontal D2R and SERT distributions contrastingly correlate with default-mode connectivity.
Although brain research has taken important strides in recent decades, the interaction and coupling of its different physiological levels is still not elucidated. Specifically, the molecular substrates of resting-state functional connectivity (rs-FC) remain poorly understood. The aim of this study was elucidating interactions between dopamine D2 receptors (D2R) and serotonin transporter (SERT) availabilities in the striatum (CPu) and medial prefrontal cortex (mPFC), two of the main dopaminergic and serotonergic projection areas, and the default-mode network. Additionally, we delineated its interaction with two other prominent resting-state networks (RSNs), the salience network (SN) and the sensorimotor network (SMN). To this extent, we performed simultaneous PET/fMRI scans in a total of 59 healthy rats using [11C]raclopride and [11C]DASB, two tracers used to image quantify D2R and SERT respectively. Edge, node and network-level rs-FC metrics were calculated for each subject and potential correlations with binding potentials (BPND) in the CPu and mPFC were evaluated. We found widespread negative associations between CPu D2R availability and all the RSNs investigated, consistent with the postulated role of the indirect basal ganglia pathway. Correlations between D2Rs in the mPFC were weaker and largely restricted to DMN connectivity. Strikingly, medial prefrontal SERT correlated both positively with anterior DMN rs-FC and negatively with rs-FC between and within the SN, SMN and the posterior DMN, underlining the complex role of serotonergic neurotransmission in this region. Here we show direct relationships between rs-FC and molecular properties of the brain as assessed by simultaneous PET/fMRI in healthy rodents. The findings in the present study contribute to the basic understanding of rs-FC by revealing associations between inter-subject variances of rs-FC and receptor and transporter availabilities. Additionally, since current therapeutic strategies typically target neurotransmitter systems with the aim of normalizing brain function, delineating associations between molecular and network-level brain properties is essential and may enhance the understanding of neuropathologies and support future drug development.


Acute subjective response to alcohol as a function of reward and punishment sensitivity.
Individual differences in subjective response to alcohol play a crucial role in the development of heavy drinking and related problems. In light of this, a growing focus of research has been identifying factors that contribute to differences in response. The aim of the present study was to determine whether individual differences in the subjective experience of rewarding and aversive effects of alcohol are a specific manifestation of general differences in reward and punishment sensitivity. Eighty-nine participants (M age=22.4, SD=1.9; 47.2% women) consumed a moderate dose of alcohol, i.e., peak breath alcohol concentration (BrAC)≈0.080g%, and rated their level of stimulation and sedation at seven timepoints over the BrAC curve. Sensitivity to reward and punishment were assessed by a self-report questionnaire prior to consumption. Multilevel growth models showed that post-consumption changes in stimulation ratings varied as a function of participants' level of reward and punishment sensitivity. Drinkers more sensitive to reward reported feeling more stimulated shortly after drinking and exhibited an attenuated rate of decline in stimulation over the blood alcohol curve, relative to drinkers with less strong reward sensitivity. Reward sensitivity was not related to subjective ratings of sedation, and punishment sensitivity was not related to either stimulation or sedation ratings. Findings suggest that reward sensitivity may increase risk for alcohol misuse among young adult social drinkers by increasing their subjective feelings of stimulation while drinking.


A novel adaptation of a parent-child observational assessment tool for appraisals and coping in children exposed to acute trauma.
Millions of children worldwide are exposed to acute potentially traumatic events (PTEs) annually. Many children and their families experience significant emotional distress and/or functional impairment following PTEs. While current research has begun to highlight a role for early appraisals and coping in promoting or preventing full recovery from PTEs, the exact nature of the relationships among appraisals, coping, and traumatic stress reactions as well as how appraisals and coping behaviors are influenced by the child's environment (e.g., parents) remains unclear; assessment tools that reach beyond self-report are needed to improve this understanding. The objective of the current study is to describe the newly created Trauma Ambiguous Situations Tool (TAST; i.e., an observational child-parent interview and discussion task that allows assessment of appraisals, coping, and parent-child processes) and to report on initial feasibility and validation of TAST implemented with child-parent dyads in which children were exposed to a PTE. As part of a larger study on the role of biopsychosocial factors in posttraumatic stress reactions, children (aged 8-13) and parents (n=25 child-parent dyads) completed the TAST during the child's hospitalization for injury. Children and parents engaged well with the TAST. The time to administer the TAST was feasible, even in a peri-trauma context. The TAST solicited a wide array of appraisals (threat and neutral) and coping solutions (proactive and avoidant). Forced-choice and open-ended appraisal assessments provided unique information. The parent-child discussion portion of the TAST allowed for direct observation of parent-child processes and demonstrated parental influence on children's appraisals and coping solutions. The TAST is a promising new research tool, which may help to explicate how parents influence their child's developing appraisals and coping solutions following a PTE. More research should examine the relationships of appraisals, coping, and parent-child processes assessed by the TAST with traumatic stress outcomes.


Memory, but not naive, peripheral blood B lymphocytes differentiate into Ig-secreting cells after CD40 ligation and costimulation with IL-4 and the differentiation factors IL-2, IL-10, and IL-3.
The ligation of CD40 on B lymphocytes by CD40 ligand, transiently expressed on activated Th cells, provides a key activation signal required for the germinal center B cell response. In vitro, human B cell activation has been investigated extensively by coculturing tonsillar B cells with CD32-transfected fibroblasts coated with anti-CD40 Abs, in the presence of cytokines (the CD40 system). When tonsillar IgD+ B cells are cultured in the CD40 system with IL-4, cells proliferate and switch to IgG, but they display a block of differentiation illustrated by the persistence of IgD expression on cycling B cells. In this study, we analyzed the responses of peripheral blood B lymphocyte fractions, which may contain fewer in vivo activated cells than those from tonsils. While the differentiation block was confirmed with peripheral naive B cells cultured in the CD40 system with IL-4, it was also observed with the combination of IL-2, IL-10, and IL-3 alone or together with IL-4 (persistence of >90% IgD+ cells, including 24-60% IgD+, IgG+ cells, and <6% IgD+, IgA+ cells after 8 days). IgD+, IgG-, and IgA- (naive) B cells secreted 70-fold less Ig than IgG+, IgA+ (memory) B cells in response to anti-CD40 plus IL-2, IL-10, and IL-3. IgG-, IgA- B cells, or IgD-, IgM+, which should include IgM+ memory cells, strongly secreted IgM, but no IgG. In conclusion, only memory B cells secreted Ig; like memory T cells, their activation requirements to differentiate into effector cells seem less stringent than those of the naive cells.


Microelectrode measurements from oxyntic cells in intact Necturus gastric mucosa.
The electrical properties of oxyntic cells were measured in intact isolated Necturus fundic mucosa by dissecting away the serosal muscle and connective tissue and impaling the oxyntic cells across their basolateral membranes. Their properties under resting [i.e., not secreting acid (10(-4) M serosal cimetidine)] and stimulated (10(-4) M histamine) conditions were compared with those of surface cells impaled across their apical membranes in a separate set of experiments. Histamine hyperpolarized the transepithelial potential by 6-10 mV and reduced the transepithelial resistance by approximately 40%. The basolateral membrane potential (Vcs) of both cell types was significantly hyperpolarized by histamine, that of oxyntic cells from a resting value of -50 to -59 mV (P less than 0.001) and that of surface cells from -50 to -54 mV (P less than 0.05). Histamine also hyperpolarized the apical membrane potential (Vmc) of the oxyntic cells; however, the Vmc of surface cells was significantly depolarized. The ratio of the apical to basolateral cell membrane resistances Ra/Rb (delta Vmc/delta Vcs resulting from transepithelial current pulses) of resting oxyntic cells was 1.1 and that of surface cells was 3.6. Stimulation did not affect the Ra/Rb of either cell type. A tenfold increase in serosal K+ concentration depolarized Vcs and increased Ra/Rb of resting and stimulated oxyntic cells, indicating a significant basolateral K+ conductance. The results are consistent with a purely passive role for surface cells and indicate that stimulation results in a simultaneous decrease of both the apical and basolateral membrane resistances of the oxyntic cells.


Attending, ignoring, and repetition: on the relation between negative priming and inhibition of return.
A series of spatial localization experiments is reported that addresses the relation between negative priming and inhibition of return. The results of Experiment 1 demonstrate that slowed responses to repeated location stimuli can be obscured by repetition priming effects involving stimulus dimensions other than spatial location. The results of Experiments 2, 3A, and 3B demonstrate that these repetition priming effects may occur only when participants are required to respond to the prime display. Together, these results suggest that differences between attended and ignored repetition effects in selective attention studies of spatial localization do not provide a basis for distinguishing between spatial negative priming and inhibition of return.


A rapid and simple electrophoretic method for the detection of mutations involving small insertion or deletion: application to beta-thalassemia.
The 1.8-kb beta-globin gene fragments of DNAs from individuals heterozygous for nine different beta-thalassemia mutations involving 1, 2, 3, 4, or 25 basepair (bp) insertions or deletions were amplified by the polymerase chain reaction (PCR). The PCR products were subjected to electrophoresis on aqueous 8% polyacrylamide gel. In each heterozygote with either a 2 to 25 bp deletion, but not with a 1 bp insertion, two slower migrating bands representing heteroduplexes in addition to the 1.8-kb homoduplex band were seen. The electrophoretic positions of these slower migrating bands were characteristic of each mutation studied. By co-amplification with known normal DNA, it was also possible to distinguish DNAs from normal individuals and from individuals who are homozygous for the small insertion/deletion mutations. These studies demonstrate that the heteroduplex formation generated in PCR can be applied as a simple method in the diagnosis of insertion/deletion mutations involving 2 to 25 bp in beta-thalassemias as well as in other genetic disorders.


Identification of traditional East Asian handmade papers through the multivariate data analysis of pyrolysis-GC/MS data.
An analytical approach based on the multivariate analysis of on-line pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) data is proposed for the identification of traditional East Asian handmade papers from different fiber material origins. This approach utilized several biomarkers detected during the Py-GC/MS analysis of paper samples. At first, the total ion chromatogram (TIC) was taken as the response and then the extracted ion chromatograms (EICs) were considered to improve the discrimination of papers. The influence of different data pretreatments (raw responses vs. normalized values) including different weightings of the variables (weighting as 1 vs. weighting as 1/STD, where STD stands for standard deviation) for principal component analysis was also investigated. The results showed that compared to the commonly used microscopy techniques, the Py-GC/MS technique proved to be able to discriminate against handmade paper materials that have similar microscopic morphologies such as Morus species vs. Broussonetia species. The data pretreatment influenced PCA modeling: the analysis based on normalized values showed more interpretable PCA group features for Moraceae species. PCA without weighting resulted unsurprisingly in discrimination through the presence of high intensity response biomarkers, while when applying weight as 1/STD, a PCA loading plot was shown to provide a group of compounds, most of them being present at low levels, to be discriminating. Additionally, the characteristic EICs can provide a data matrix for statistical analysis avoiding the interference from a co-eluting compound and background compared to the data matrix obtained from the TIC. As a result, a quick Py-GC/MS based handmade paper identification procedure using PCA modeling of the characteristic EICs was proposed for the first time in the identification of traditional East Asian handmade papers. This procedure could be very beneficial for cultural heritage applications.


Subcortical neural synchrony and absolute thresholds predict frequency discrimination independently.
The neural mechanisms of pitch coding have been debated for more than a century. The two main mechanisms are coding based on the profiles of neural firing rates across auditory nerve fibers with different characteristic frequencies (place-rate coding), and coding based on the phase-locked temporal pattern of neural firing (temporal coding). Phase locking precision can be partly assessed by recording the frequency-following response (FFR), a scalp-recorded electrophysiological response that reflects synchronous activity in subcortical neurons. Although features of the FFR have been widely used as indices of pitch coding acuity, only a handful of studies have directly investigated the relation between the FFR and behavioral pitch judgments. Furthermore, the contribution of degraded neural synchrony (as indexed by the FFR) to the pitch perception impairments of older listeners and those with hearing loss is not well known. Here, the relation between the FFR and pure-tone frequency discrimination was investigated in listeners with a wide range of ages and absolute thresholds, to assess the respective contributions of subcortical neural synchrony and other age-related and hearing loss-related mechanisms to frequency discrimination performance. FFR measures of neural synchrony and absolute thresholds independently contributed to frequency discrimination performance. Age alone, i.e., once the effect of subcortical neural synchrony measures or absolute thresholds had been partialed out, did not contribute to frequency discrimination. Overall, the results suggest that frequency discrimination of pure tones may depend both on phase locking precision and on separate mechanisms affected in hearing loss.


Genetically Predicted Glucose-Dependent Insulinotropic Polypeptide (GIP) Levels and Cardiovascular Disease Risk Are Driven by Distinct Causal Variants in the GIPR Region.
There is considerable interest in GIPR agonism to enhance the insulinotropic and extrapancreatic effects of GIP, thereby improving glycemic and weight control in type 2 diabetes (T2D) and obesity. Recent genetic epidemiological evidence has implicated higher GIPR-mediated GIP levels in raising coronary artery disease (CAD) risk, a potential safety concern for GIPR agonism. We therefore aimed to quantitatively assess whether the association between higher GIPR-mediated fasting GIP levels and CAD risk is mediated via GIPR or is instead the result of linkage disequilibrium (LD) confounding between variants at the GIPR locus. Using Bayesian multitrait colocalization, we identified a GIPR missense variant, rs1800437 (G allele; E354), as the putatively causal variant shared among fasting GIP levels, glycemic traits, and adiposity-related traits (posterior probability for colocalization [PPcoloc] > 0.97; PP explained by the candidate variant [PPexplained] = 1) that was independent from a cluster of CAD and lipid traits driven by a known missense variant in APOE (rs7412; distance to E354 ∼770 Kb; R 2 with E354 = 0.004; PPcoloc > 0.99; PPexplained = 1). Further, conditioning the association between E354 and CAD on the residual LD with rs7412, we observed slight attenuation in association, but it remained significant (odds ratio [OR] per copy of E354 after adjustment 1.03; 95% CI 1.02, 1.04; P = 0.003). Instead, E354's association with CAD was completely attenuated when conditioning on an additional established CAD signal, rs1964272 (R 2 with E354 = 0.27), an intronic variant in SNRPD2 (OR for E354 after adjustment for rs1964272: 1.01; 95% CI 0.99, 1.03; P = 0.06). We demonstrate that associations with GIP and anthropometric and glycemic traits are driven by genetic signals distinct from those driving CAD and lipid traits in the GIPR region and that higher E354-mediated fasting GIP levels are not associated with CAD risk. These findings provide evidence that the inclusion of GIPR agonism in dual GIPR/GLP1R agonists could potentiate the protective effect of GLP-1 agonists on diabetes without undue CAD risk, an aspect that has yet to be assessed in clinical trials.