Insufficient Inhibition Research Articles

Heart-rate-variability (HRV), predicts outcomes in COVID-19.

BackgroundPatients with COVID-19 present with a variety of clinical manifestations, ranging from mild or asymptomatic disease to severe illness and death. Whilst previous studies have clarified these and several other aspects of COVID-19, one of the ongoing challenges regarding COVID-19 is to determine which patients are at risk of adverse outcomes of COVID-19 infection. It is hypothesized that this is the result of insufficient inhibition of the immune response, with the vagus nerve being an important neuro-immuno-modulator of inflammation. Vagus nerve activity can be non-invasively indexed by heart-rate-variability (HRV). Therefore, we aimed to assess the prognostic value of HRV, as a surrogate marker for vagus nerve activity, in predicting mortality and intensive care unit (ICU) referral, in patients hospitalized with COVID-19.MethodsA retrospective cohort study including all consecutive patients (n = 271) diagnosed and hospitalized with COVID-19 between March 2020 and May 2020, without a history of cardiac arrhythmias (including atrial and ventricular premature contractions), pacemaker, or current bradycardia (heart rate <50 bpm) or tachycardia (heart rate >110 bpm). HRV was based on one 10s ECG recorded at admission. 3-week survival and ICU referral were examined.ResultsHRV indexed as standard deviation of normal to normal heartbeat intervals (SDNN) predicted survival (H.R. = 0.53 95%CI: 0.31–0.92). This protective role was observed only in patients aged 70 years and older, not in younger patients. HRV below median value also predicted ICU referral within the first week of hospitalization (H.R = 0.51, 95%CI: 0.29–0.90, P = 0.021).ConclusionHigher HRV predicts greater chances of survival, especially in patients aged 70 years and older with COVID-19, independent of major prognostic factors. Low HRV predicts ICU indication and admission in the first week after hospitalization.

Retinal degeneration-3 protein attenuates photoreceptor degeneration in transgenic mice expressing dominant mutation of human retinal guanylyl cyclase

Different forms of photoreceptor degeneration cause blindness. Retinal degeneration-3 protein (RD3) deficiency in photoreceptors leads to recessive congenital blindness. We proposed that aberrant activation of the retinal membrane guanylyl cyclase (RetGC) by its calcium-sensor proteins (guanylyl cyclase–activating protein [GCAP]) causes this retinal degeneration and that RD3 protects photoreceptors by preventing such activation. We here present in vivo evidence that RD3 protects photoreceptors by suppressing activation of both RetGC1 and RetGC2 isozymes. We further suggested that insufficient inhibition of RetGC by RD3 could contribute to some dominant forms of retinal degeneration. The R838S substitution in RetGC1 that causes autosomal-dominant cone–rod dystrophy 6, not only impedes deceleration of RetGC1 activity by Ca2+GCAPs but also elevates this isozyme's resistance to inhibition by RD3. We found that RD3 prolongs the survival of photoreceptors in transgenic mice harboring human R838S RetGC1 (R838S+). Overexpression of GFP-tagged human RD3 did not improve the calcium sensitivity of cGMP production in R838S+ retinas but slowed the progression of retinal blindness and photoreceptor degeneration. Fluorescence of the GFP-tagged RD3 in the retina only partially overlapped with immunofluorescence of RetGC1 or GCAP1, indicating that RD3 separates from the enzyme before the RetGC1:GCAP1 complex is formed in the photoreceptor outer segment. Most importantly, our in vivo results indicate that, in addition to the abnormal Ca2+ sensitivity of R838S RetGC1 in the outer segment, the mutated RetGC1 becomes resistant to inhibition by RD3 in a different cellular compartment(s) and suggest that RD3 overexpression could be utilized to reduce the severity of cone–rod dystrophy 6 pathology.

Atypical HUS and Crohn\u2019s disease\u2014interference of intestinal disease activity with complement-blocking treatment

BackgroundIn atypical hemolytic-uremic syndrome (aHUS), various defects of the complement system have been reported to explain pathophysiology. Therapeutic options for complement inhibition are well-recognized; however, the links between various immune-derived diseases and aHUS are unclear, and their interference with treatment efficacy during long-term complement-blocking therapy is scarcely known.Case-diagnosis/treatmentWe present a pediatric patient who developed aHUS with acute kidney injury in parallel with the onset of Crohn’s disease (CD), and who required long-term complement-blocking therapy with eculizumab (ECU). Unexpectedly, during the 6-year ECU treatment, an important intra-patient variation of the degree of complement inhibition was observed. In spite of continuous and stable doses of complement-blocking therapy, periods of incomplete blockade were observed in strong association with relapses of CD. When conventional and later biological therapy with adalimumab was introduced, with CD going into remission, complement blockade became complete again. Despite periodically low ECU levels and insufficient complement inhibition, no clinical or hematological signs of aHUS recurrence were detected during CD relapses.ConclusionIn aHUS cases secondary to CD, close monitoring of both complement inhibition and serum ECU levels is needed as intestinal disease can interfere with complement-blocking treatment. Increased doses of ECU may be necessary to maintain therapeutic blood levels of ECU and full complement blockade, especially if the intestinal disease is not under control.

Low Dose Aspirin in high-risk pregnancies: The volatile effect of acetylsalicylic acid on the inhibition of platelets uncovered by G. Born’s light transmission aggregometry

Preeclampsia still represents a life-threatening pregnancy complication, associated with severe maternal and neonatal morbidity and mortality. Low-dose Aspirin is advised to avoid preeclampsia in high-risk pregnancies worldwide. As Aspirin does not cover all women at risk, the prescription raises questions concerning optimal target population, dosage, and onset of therapy.The aim of this study was to test platelet responsiveness on Aspirin by optical aggegrometry, to gain robust biochemically assessment data of Aspirin in an obstetric cohort. 248 women at high risk for development of preeclampsia were included in the study. Aspirin-prophylaxis was administered either in 100 mg (n = 229) or 150 mg (n = 90) daily. Dosing of 100 mg Aspirin was maintained if testing revealed a sufficient platelet inhibition. If platelet inhibition was insufficient, dosage was increased to 150 mg Aspirin and re-testing was advised. 91 patients (91/229 = 39.7%) presented a sufficient inhibitory Aspirin effect at a dosage of 100 mg, but in 138 patients LTA showed an inadequate Aspirin response (138/229 = 60.3%). In 19 women 150 mg Aspirin was administered as starting dose due to new recommendations. Of all women at 150 mg Aspirin 64 did not properly respond (35.4%). The overall rate of sufficient responding women regardless the Aspirin dose was 64.6%.This study demonstrates still an insufficient inhibition of platelet aggregation in about 1/3 of women even with a dosage of 150 mg Aspirin daily, who might potentially benefit from further increase. These data show, that there is a need for further research to allow a personalized approach for individualized Aspirin therapy, maximizing the preventive benefit for mother and child.

ANalgesic Efficacy and safety of MOrphiNe versus methoxyflurane in patients with acute myocardial infarction: the rationale and design of the ANEMON-SIRIO 3 study: a multicentre, open-label, phase II, randomised clinical trial

IntroductionThe unfavourable influence of morphine on the pharmacokinetics of ticagrelor resulting in weaker and retarded antiplatelet effect in patients with acute coronary syndrome (ACS) has been previously shown. Replacing morphine...

Exploring diverse sorghum ( Sorghum bicolor (L.) Moench) accessions for malt amylase activity

Sorghum is a climate resilient grain cereal crop originating from Africa that has traditionally been used to make a variety of fermented beverages such as pito and baijiu. In Western markets, the use of sorghum to produce beers and beverages has recently risen due to the visibility of a gluten sensitive/intolerant market and a growing interest in unique inputs for beverage production. As such, there is a developing body of research on sorghum as a malted input into beverages. A major limitation to the wider adoption of sorghum as a substrate in mashing is its low activity of amylolytic enzymes, either the result of insufficient activity or inhibition by endogenous compounds. A collection of 42 diverse accessions representing the grain sorghum diversity panel, was evaluated for associations between alpha and beta amylase content, race, origin, and seed colour as well as two classes of amylase inhibitors, phenols and tannins. Among these accessions are several commonly used genetic resources, including reference line BTx623. Notable findings include accessions with high alpha amylase content, sources that may harbour additional high amylase sorghums, associations with grain colour, and populations which may be used to genetically map the trait. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling

Oscillations of C-peptide in the euglycemic clamp and their effect on the pharmacodynamic assessment of insulin preparations.

C-peptide should be continuously suppressed. However, increased postdosing C-peptide is not an uncommon phenomenon in euglycemic clamp studies involving healthy participants. This study aimed to determine the extent to which the postdosing C-peptide increases from the baseline that could affect the accuracy of glucodynamics in euglycemic clamp studies involving healthy subjects. First, 10 healthy males underwent a 10-h euglycemic clamp without exogenous insulin administration to obtain a reference interval (RI) for the ratio of C-peptide after 0min (CPt ) to baseline C-peptide (CP0 ). Then, the data from a pharmacokinetic and pharmacodynamic study of insulin aspart (IAsp) were analyzed, and 70 eligible clamps were grouped by CPt /CP0 : group A ([CPt /CP0 ]max >upper limit of RI), group B (1<[CPt /CP0 ]max ≤upper limit of RI), and group C ([CPt /CP0 ]max ≤1). The differences in basal and clamped blood glucose, CPt /CP0 , and the pharmacokinetics and pharmacodynamics of IAsp were compared, and the relationship between elevated CPt and the accuracy of pharmacodynamics was analyzed. The RI of CPt /CP0 was 22.7%-152.1%; 1.5×baseline might be a ceiling for the increase in CPt under stable conditions. The maximum glucose infusion rate (GIR) in group A tended to be higher than that in group B or C (Pfor trend =0.033). The AUCGIR,0-10h in groups A, B, and C was 1983±650,1682±454, and 1479±440mg/kg (P=0.047), respectively, under comparable IAsp exposure. No intergroup difference was detected in clamped glucose, IAsp dose, or body mass index. In conclusion, postdosing C-peptide over 1.5× baseline indicates insufficient inhibition of endogenous insulin secretion, which could compromise the pharmacodynamics of insulin preparations.

White matter integrity moderates the relation between experienced childhood maltreatment and fathers' behavioral response to infant crying.

The ability to provide appropriate responses to infant distress is vital to paternal care, but may be affected by fathers’ experiences of childhood maltreatment. Detrimental effects of childhood maltreatment have been found in the adult brain's white matter fibers, accompanied with impaired emotional and cognitive functioning. In the current study (N = 121), we examined new and expectant fathers’ childhood maltreatment experiences (i.e. emotional and physical abuse and neglect), current behavioral responses (i.e. handgrip force) to infant cry sounds, and white matter integrity using diffusion tensor imaging. First, more exposure to childhood maltreatment was associated with more use of excessive handgrip force in response to infant crying by fathers. Second, the association between experienced childhood maltreatment and white matter integrity was not significant in whole‐brain analyses. Lastly, we found that the association between maltreatment exposure and excessive handgrip force during infant crying was absent in fathers with high tract integrity in the bilateral uncinate fasciculus. These findings possibly point to insufficient behavioral inhibition or emotional dysregulation in fathers who experienced childhood maltreatment, but buffering for this effect in those with larger integrity in brain fibers connecting the amygdala and prefrontal cortex.

Dysfunction of Orbitofrontal GABAergic Interneurons Leads to Impaired Reversal Learning in a Mouse Model of Obsessive-Compulsive Disorder

Cognitive inflexibility is a cardinal symptom of obsessive-compulsive disorder (OCD) and often manifests as impaired reversal learning. Abnormal recruitment of the orbitofrontal cortex (OFC)-striatal circuit is implicated in reversal learning deficits in patients with OCD. However, the precise circuitry mechanism underlying normal and impaired reversal learning remains elusive. Using fiber photometry and optogenetics, we demonstrated cell-type-specific activity dynamics in the OFC-striatal circuit underlying normal reversal learning and cell-type-specific dysfunctions that causally lead to impaired reversal learning in an OCD mouse model (Sapap3 knockout mice). After contingency reversal, OFC GABAergic interneurons increase the activity in response to previously rewarded but currently non-reward cues to inhibit the elevated activity of OFC excitatory neurons encoding inappropriate cue-reward association. Striatal direct-pathway medium spiny neurons (D1-MSNs) gradually re-establish their response preference for rewarded versus non-reward cues. These activity dynamics together mediated normal reversal learning. In Sapap3 knockout OCD mouse model, the increase in activity of OFC GABAergic interneurons in response to previously rewarded but currently non-reward cues after contingency reversal was reduced, which resulted in insufficient inhibition on OFC excitatory neurons, which in turn led to a more severe inversion of the response preference of D1-MSNs for rewarded versus non-reward cues, ultimately resulting in slower reversal learning. These dysfunctions were causally involved in reversal learning impairments. Our findings identified OFC GABAergic interneurons as the key therapeutic target to treat cognitive inflexibility in OCD and may be generally applicable to cognitive inflexibility in other neuropsychiatric disorders.

Dual synaptic inhibitions of brainstem neurons by GABA and glycine with impact on Rett syndrome.

Rett syndrome (RTT) is a neurodevelopmental disease caused mostly by mutations in the MECP2 gene. People with RTT show breathing dysfunction attributable to the high rate of sudden death. Previous studies have shown that insufficient GABA synaptic inhibition contributes to the breathing abnormalities in mouse models of RTT, while it remains elusive how the glycine system is affected. We found that optogenetic stimulation of GAD-expressing neurons in mice produced GABAergic and glycinergic postsynaptic inhibitions of neurons in the hypoglossal nucleus (XII) and the dorsal motor nucleus of vagus (DMNV). By sequential applications of bicuculline and strychnine, such inhibition appeared approximately 44% GABAA ergic and 52% glycinergic in XII neurons, and approximately 49% GABAA ergic and 46% glycinergic in DMNV neurons. Miniature inhibitory postsynaptic potentials (mIPSCs) in these neurons were approximately 47% GABAA ergic and 49% glycinergic in XII neurons, and approximately 48% versus 50% in DMNV neurons, respectively. Consistent with the data, our single-cell polymerase chain reaction studies indicated that transcripts of GABAA receptor γ2 subunit (GABAA Rγ2) and glycine receptor β subunit (GlyRβ) were simultaneously expressed in these cells. In MeCP2R168X mice, proportions of GABAA ergic and glycinergic mIPSCs became approximately 28% versus 69% in XII neurons, and approximately 31% versus 66% in DMNV cells. In comparison with control mice, the GABAA ergic and glycinergic mIPSCs decreased significantly in the XII and DMNV neurons from the MeCP2R168X mice, so did the transcripts of GABAA Rγ2 and GlyRβ. These results suggest that XII and DMNV neurons adopt dual GABAA ergic and glycinergic synaptic inhibitions, and with Mecp2 disruption these neurons rely more on glycinergic synaptic inhibition.

DDRE-01. ACQUIRED AND INTRINSIC RESISTANCE TO VEMURAFENIB IN BRAFV600E-DRIVEN MELANOMA BRAIN METASTASES

Abstract BRAF V600-mutated melanoma brain metastases (MBMs) are responsive to BRAF inhibitors, but clinical responses are less durable than those of extracranial metastases. We studied the impact of the drug efflux proteins P-glycoprotein (P-gp; ABCB1) and breast cancer resistance protein (BCRP; ABCG2) at the blood-brain barrier (BBB) on the efficacy of vemurafenib against BRAFV600E-mutated A375 MBMs. We intracranially implanted A375 tumor cells in wild-type and Abcb1a/b;Abcg2-/- mice. We characterized the tumor BBB, analyzed drug levels in plasma and brain lesions after oral vemurafenib and determined the efficacy against brain metastases and subcutaneous lesions. MRI shows that A375 MBMs disrupt BBB integrity, but vemurafenib accumulation in MBMs was still reduced by P-gp/BCRP. Vemurafenib is also less efficacious against MBMs in wild-type mice compared to Abcb1a/b;Abcg2-/- mice. Vemurafenib efficacy against subcutaneous A375 tumors was similar in both strains. Even in Abcb1a/b;Abcg2-/- mice, A375 MBMs rapidly developed resistance, which was unrelated to pharmacokinetic issues or insufficient inhibition of MAPK/PI3K pathways. Taken together, these studies demonstrate that although the BBB is disrupted in MBMs, P-gp/BCRP still limit the efficacy of vemurafenib. Moreover, the response to vemurafenib is less and of shorter duration also due to rapidly acquired resistance, most likely by resorting to non-canonical growth signaling.

Divergent Roles of PI3K Isoforms in PTEN-Deficient Glioblastomas

SUMMARYLoss of PTEN, the negative regulator of PI3K activity, is frequent in glioblastomas (GBMs). However, the role of the two major PI3K isoforms, p110α and p110β, in PTEN-deficient gliomagenesis remains unknown. We show that PTEN-deficient GBM largely depends on p110α for proliferation and p110β for migration. Genetic ablation of either isoform delays tumor progression in mice, but only ablating both isoforms completely blocks GBM driven by the concurrent ablation of Pten and p53. BKM120 (buparlisib) treatment only modestly prolongs survival in mice bearing intracranial Pten/p53 null tumors due to partial pathway inhibition. BKM120 extends the survival of mice bearing intracranial tumors in which p110β, but not p110α, has been genetically ablated in the Pten/p53 null glioma, indicating that BKM120 fails to inhibit p110β effectively. Our study suggests that the failure of PI3K inhibitors in GBM may be due to insufficient inhibition of p110β and indicates a need to develop brain-penetrant p110α/β inhibitors.

Decreased HLA-C1 alleles in couples of KIR2DL2 positive women with recurrent pregnancy loss

Specific killer cell immunoglobulin-like receptor (KIR) in women with recurrent pregnancy loss (RPL) and HLA ligands in couples invoke a susceptibility to RPL. However, the relationship between KIR2DL2 and its cognate ligand HLA-C1 has not been explored. In this prospective cohort study, 160 Caucasian women with RPL and 99 partners were included. KIR/HLA-C typing, NK assay, Th1/Th2 intracellular cytokine ratios, 25-(OH)-vitamin D level, and the presence of autoantibodies were analyzed. KIR2DL2 positive women (P = 0.023) and their partners (P = 0.017) had lower allele frequencies of HLA-C1 than those of KIR2DL2 negative women. KIR2DL2 positive women had significantly lower genotype frequency of HLA-C1C1 as compared to the North American Caucasian population controls (P < 0.05). In the partners of KIR2DL2 positive women, there was a substantially higher frequency of HLA-C2C2 than controls (P = 0.016). Besides, KIR2DL2 negative women had a higher prevalence of anti-ssDNA antibody as compared with that of KIR2DL2 positive women (P = 0.043). There were no differences in the distribution of HLA-C genotypes based on KIR2DL2, regardless of pregnancy outcome in women with RPL and their partners while on immunomodulation treatment. In conclusion, decreased ligands for inhibitory KIRs (inhKIR) could lead to insufficient inhibition of maternal uterine NK cells toward the trophoblast, thereby contributing to the pathogenesis of RPL. Specific KIR and HLA-C genotyping may predict the reproductive outcome of women with RPL.

Abstract 5182: Examining the effects of honokiol on EGFR-positive glioblastoma multiforme

Abstract Glioblastoma (GBM) is a highly aggressive and lethal form of primary brain tumor. Systemic suppression of the immune system, specifically T-cell impairment, significantly contributes to GBM development and progression. Recent advances in cancer immunotherapies have shown great promise in alleviating such tumor-associated immunosuppression, namely programmed cell death 1 (PD-1)/programmed death ligand 1 (PD-L1) checkpoint inhibitors. Regrettably, early clinical studies evaluating the efficacy of checkpoint inhibition in recurrent GBM have rendered only modest improvements in overall survival rates, due partially to insufficient inhibition of AKT-STAT3 pathways which are often dysregulated downstream of hyperactive epidermal growth factor receptors (EGFR) in GBM. In the current study, we aim to evaluate a newly discovered small molecule inhibitor of AKT-STAT3, honokiol, and its ability to suppress AKT-STAT3 activity in the context of hyperactive EGFR in GBM. Using U87-MG cells that stably overexpress a common hyperactivating mutation of EGFR, EGFRvIII, we have preliminary evidence that honokiol is an effective inhibitor of U87-EGFRvIII cellular migration as a result of its robust AKT-STAT3 pathway suppression. Thus, honokiol may provide a novel means of pharmacological inhibition of AKT-STAT3 hyperactivity in GBM and therefore greater clinical efficacy of immune checkpoint inhibitors in this deadly disease. Citation Format: Keller Smith, Blake Johnson. Examining the effects of honokiol on EGFR-positive glioblastoma multiforme [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5182.

Synthesis, characterization, antioxidant and antidiabetic studies of Cu(II) and Zn(II) complexes of tolfenamic acid/mefenamic acid with 1-methylimidazole

Cu(II) and Zn(II) complexes of tolfenamic acid (tolf) and mefenamic acid (mef) with 1-methylimidazole (1-mdz) formulated as [Cu(tolf)2(1-mdz)2] (1) [Cu(mef)2(1-mdz)2]·H2O (2) and [Zn2(tolf)4(1-mdz)2] (3) were synthesized and characterized by elemental analysis, spectroscopic techniques and single crystal X-ray crystallography. The molecular structure of the Cu(II) complexes, [Cu(tolf)2(1-mdz)2] (1) and [Cu(mef)2(1-mdz)2]·H2O (2) revealed CuN2O4 distorted octahedral geometry while [Zn2(tolf)4(1-mdz)2] (3) crystallized as dinuclear with Zn2N2O8 form and the geometry around each Zn(II) ion is square pyramidal. EPR spectra of 1 and 2 confirmed paramagnetic character with exchange interaction index, G of 5.27 and 7.39 respectively. The g tensor value, g// < 2.3 for 1 and 2 indicate covalent character supported by their non-electrolytic nature (Molar conductivity, Ʌm, 11.30–15.65 µS cm−1). The scavenging and iron chelating potentials of the compounds were evaluated. The compounds exhibit good antioxidant activity and SOD-mimetic activity determined for the complexes is in the order 3 ˃ 2˃ 1, the IC50 values for 2 and 3 being <1.00 µM are positive antioxidant indices, and are comparable with that of the native bovine, Cu,Zn-SOD (IC50 = 0.48 µM). A dismal performance observed for the dose-dependent antidiabetic potentials against alpha-amylase and alpha-glucosidase is attributed to insufficient inhibition of the latter.

Emerging Role of Fentanyl in Antiplatelet Therapy.

Fentanyl is a potent synthetic opioid used to alleviate severe and chronic pain, as well as an adjunct to general or local anesthesia. Although fentanyl has been used for decades, its full effects are still unknown. Its analgesic and anesthetic activity arises from the stimulation of μ-opioid receptors, resulting in the inhibition of adenyl cyclase and downregulation of cyclic adenosine 3',5'-monophosphate (cAMP), as well as decreased calcium channel activity and increased potassium channel activity. The μ-opioid receptors are abundantly distributed within the central nervous system, where they mediate analgesia, and in the nerve cells of the intestines, where they regulate gastrointestinal tract motility in the secretion or transport of fluids and electrolytes. They are also expressed in blood cells, blood vessel cells, and skin. Given the widespread distribution of μ-opioid receptors, it is likely that fentanyl may also regulate the activity of many other cells, including platelets. Recent findings indicate that it may impair the action of ticagrelor: an oral P2Y12 receptor inhibitor acting as an antiplatelet drug. It could pose a risk of insufficient platelet inhibition and result in thrombotic complications in patients with coronary artery disease. This article tackles the issue of fentanyl interactions with antiplatelet drugs. The mechanism of this phenomenon is not fully understood. Similarly, the biological effects exerted by fentanyl on platelets and the presence of opioid receptors on the platelet surface remain an open question.

M5. A MULTIMODAL STUDY ON THE INFLUENCE OF LEFT-BEHIND CHILDREN’S VISUAL PERCEPTION PROCESSING

BackgroundLeft-behind children（LBC has experienced parent-child separation, which is a special parent-child separation and social isolation model. Adverse events in early development may have important effects on the development of the nervous system, including the influence on sensory and perceptual cognitive processing, thus increasing the susceptibility to mental illness. Meanwhile, there are various neurological and brain root causes behind left-behind children’s learning difficulties, non-socialization and other psychological behavior problems. In addition, left-behind may have sensory, perceptual and cognitive processing abnormalities. This study tries to elaborate and extract the eye movement characteristics of left-behind children, lay a foundation for the discussion of the mechanism of abnormal visual processing, and provide a scientific basis for follow-up of left-behind children and the early identification, prevention and control of mental diseases.MethodsThe standards for grouping left-behind children are as follows: the children who are separated from both parents 6 months after birth and spend less than 2 months with their parents each month, and the non-left-behind children with matched gender, age and education years, exclusion criteria is: neurological diseases, family history of mental illness, broken family (single parent, bereavement), high myopia (greater than 1000 degrees), eye and eyelash cannot be identified by infrared light. EyeControl portable eye movement tester (built-in infrared, speed: 100Hz, error <0.5°) developed by Qingtech and its software processing system were used. The subjects’ eye movements and various analysis indexes in various tasks were collected. The eye movement indexes of smooth pursuit eye movement, fixation stability and free browsing were collected, so as to investigate the effects of left-behind on static attention retention, visual tracking and face recognition in social scenes.ResultsAmong the 175 children, the smooth tracking mean deviation of LBC is significantly increased (P=0.017). The mean deviation of the O and S-curve tracking of the LBC is significantly increased (P=0.018; P=0.034), and the OS curve mean deviation and hyperactivity correlated (r=-0.273, P=0.011). In terms of the time that peak deviation takes (F=4.329, P=0.014), there is significant interaction between the age groups and the left-behind groups. In the social scenes, the first fixation time of face (P=0.039), the number of eye entry count (P=0.020), and the eye fixation time (P=0.003) of LBC are all significantly reduced, and the number of mouth entry count (P=0.007) and the number of fixation (P=0.023) of LBC are all significantly increased. Under the circumstance of low facial processing, there is a significant difference between LBC and NLBC (P=0.009), and that difference is not significant under the circumstance of high facial processing, and there is a significant difference between low facial processing LBC and high facial processing NLBC (P=0.007).DiscussionLeft-behind children have obstacles in dynamic visual information processing, insufficient inhibition and control, and poor cognitive flexibility, which may be related to hyperactivity. The attention retention in visual tracking decreased with the increase of age. Left-behind children have difficulties in face recognition in social scenes. In the case of low facial processing, left-behind children aggravated the defects of speech comprehension, while in the case of high facial processing, left-behind children had no significant influence on speech comprehension. Therefore, we should intervene the left-behind children in the early days, and this study provides a scientific basis for the formulation of social policies.

Deciphering ion transporters, kinases and PDZ-adaptor molecules that mediate guanylate cyclase C agonist-dependent intestinal fluid loss in vivo

BackgroundThe molecular basis for heat-stable Escherichia coli enterotoxin (STa) action and its synthetic analogue linaclotide is well understood at the enterocyte level. Pharmacologic strategies to prevent STa-induced intestinal fluid loss by inhibiting its effector molecules, however, have achieved insufficient inhibition in vivo. Aims and experimental approachTo investigate whether the currently discussed effector molecules and signaling mechanisms of STa/linaclotide-induced diarrhea have similar relevance in vivo than at the enterocyte level, we studied the effect of 10−7M of the STa analogue linaclotide on short circuit current (Isc) of chambered isolated jejunal mucosa, and on the in vivo action on fluid transport in a perfused segment of proximal jejunum of anesthetized mice. The selected mice were deficient of transport (NHE3, CFTR, Slc26a3/a6), adaptor (NHERF1-3), or signal transduction molecules [cGMP-dependent kinase II (GKII)] considered to be downstream effectors after STa/linaclotide binding to guanylate cyclase C (GCC). Selective NHE3 inhibition by tenapanor was also employed. Key results, conclusions and implicationsThe comparison allowed the separation of effectors for stimulation of electrogenic anion secretion and for inhibition of electrolyte/fluid absorption in response to STa/linaclotide. The cGKII-NHERF1-CFTR and cGKII-NHERF2-NHE3 interactions are indeed major effectors of small intestinal fluid loss downstream of GCC activation in vitro and in vivo, but 50% of the linaclotide-induced fluid loss in vivo, while dependent on CFTR activation and NHE3 inhibition, does not involve cGKII, and 30% does not depend on NHERF1 or NHERF2. A combined NHERF1 and NHERF2 inhibition appears nevertheless a good pharmacological strategy against STa-mediated fluid loss.

Intensified P2Y12 inhibition for high-on treatment platelet reactivity

High on treatment platelet reactivity (HPR) during treatment with clopidogrel has been consistently found to be strong risk factor for recurrent ischemic events after percutaneous coronary intervention (PCI). Insufficient P2Y12 receptor inhibition contributes to HPR measured by the VerifyNow (VN) assay. Prasugrel and ticagrelor are more potent P2Y12 inhibitors than clopidogrel and commonly substituted for clopidogrel when HPR is documented, however benefit of VN guided intensified antiplatelet therapy is uncertain. We identified patients who had undergone platelet reactivity testing after PCI with VN after pretreatment with clopidogrel (n = 252) in a single center observational analysis. Patients who had HPR defined as PRU > 208 were switched to alternate P2Y12 inhibitors. Primary clinical endpoint was 1-year post PCI combined cardiovascular death, myocardial infarction (MI), and stent thrombosis. One hundred and eight (43%) subjects had HPR and were switched to prasugrel (n = 60) and ticagrelor (n = 48). Risk of recurrent 1-year primary endpoint remained higher for HPR patients switched to either ticagrelor or prasugrel as compared to subjects who had low on treatment platelet reactivity (n = 144) (LPR) on clopidogrel [Hazard Ratio: 3.5 (95% CI 1.1-11.1); p = 0.036)]. Propensity score matched analysis demonstrated higher event rates in patients with HPR on alternate P2Y12 inhibitor as compared to patients with LPR (log-rank: p = 0.044). The increased risk of recurrent events associated with HPR measured by VN is not completely attenuated by switching to more potent P2Y12 inhibitors. Non-P2Y12 mediated pathways likely contribute to increased incidence of thrombotic events after PCI in subjects with HPR.

Effect of mycophenolic acid on inosine monophosphate dehydrogenase (IMPDH) activity in liver transplant patients

Due to the development of immunosuppressants, the focus in transplanted patients has shifted from short-term to long-term survival as well as a better adjustment of these drugs in order to prevent over- and under-immunosuppression. Mycophenolic acid (MPA) is a noncompetitive inhibitor of inosine monophosphate dehydrogenase (IMPDH) and approved for prophylaxis of acute rejection after kidney, heart, and liver transplantation, where it has become a part of the standard therapy. Targeting inosine monophosphate IMPDH activity as a surrogate pharmacodynamic marker of MPA-induced immunosuppression may allow a more accurate assessment of efficacy and aid in limiting toxicity in liver transplanted patients. Assess IMPDH-inhibition in liver transplant recipients and its impact on biliary/infectious complications, acute cellular rejection (ACR) and liver dependent survival. This observational cohort study comprises 117 liver transplanted patients that were treated with mycophenolate mofetil (MMF) for at least 3 months. Blood samples (BS) were collected and MPA serum level and IMPDH activity were measured before (t(0)), 30minutes (t(30)) and 2h after (t(120)) MMF morning dose administration. Regarding MPA, we assessed the area under the curve (AUC). Patients were prospectively followed up for one year and assessed for infectious and biliary complications, episodes of ACR and liver dependent survival. The MPA levels showed a broad interindividual variability at t(0) (2.0±1.8ng/ml), t(30) (12.7±9.0ng/ml) and t(120) (7.5±4.3ng/ml). Corresponding IMPDH activity was at t(o) (23.2±9.5 nmol/h/mg), at t(30) (16.3±8.8 nmol/h/mg) and t(120) (18.2±8.7 nmol/h/mg). With regard to MPA level we found no correlation with infectious or biliary complications within the follow-up period. Patients with baseline IMPDH(a) below the median had significant more viral infections (6 (10.2%) vs. 17 (29.3%); P=0.009) with especially more cytomegalovirus (CMV) infections (1 (3.4%) vs. 6 (21.4%); P=0.03)). Furthermore, patients with baseline IMPDH(a) above the median developed more often non-anastomotic biliary strictures (8 (13.6%) vs. 1 (1.7%), P=0.03). We found the group reaching the combined clinical endpoint of death and re-transplantation showing significantly lower MPA baseline values (t(0) 0.9±0.7 vs. 2.1±1.8μg/ml Mann-Whitney-U: P=0.02). We calculated a simplified MPA(AUC) with the MPA level at baseline, 30 and 120minutes after MPA administration. Whereas we found no differences with regard to baseline characteristics at entry into the study patients with MPA (AUC) below the median experienced significantly more often the combined clinical endpoint (12.1% (7/58) vs. 0.0% (0/57); P=0.002) and had a reduced actuarial re-transplantation-free survival (1.0 year vs. 0.58 years; Log-rank: P=0.007) during the prospective one-year follow-up period. In univariate and multivariate analysis including gender, age, BMI, ACR, MPA (AUC) and IMPDH(a) only BMI, MPA (AUC) and IMPDH(a) were independently associated with reduced actuarial re-transplantation-free survival. MPA-levels and IMPDH-activity in liver transplanted patients allows individual risk assessment. Patients with higher IMPDH inhibition acquire more often viral infections. Insufficient IMPDH inhibition is associated with development of non-anastomotic bile duct strictures and reduced re-transplantation-free survival.