Fe3O4@p-Naphtholbenzein as a novel nano-sorbent for highly effective removal and recovery of Berberine: Response surface methodology for optimization of ultrasound assisted dispersive magnetic solid phase extraction.
A hydrophobic surface modified iron oxide magnetic nano-sorbent was successfully synthesized, characterized and utilized for highly effective removal and recovery of Berberine (Brb) by Ultrasound Assisted Dispersive Magnetic Solid Phase Extraction (UADM-SPE). Fabrication of p-Naphtholbenzein-coated magnetic nanoparticles (PNB-MNPs) was certified by characteristic analyses such as: Fourier transform infrared spectroscopy (FT-IR), X-ray Diffraction (XRD), thermogravimetric analysis (TGA), vibrating sample magnetometer (VSM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The characterized PNB-MNPs were utilized to develop a UADM-SPE method for Brb, a well-known isoquinoline alkaloid with a highlighted chemical skeleton for designing selective and powerful bioactive molecules. Affecting parameters on the extraction of Brb were optimized through an orthogonal rotatable central composite design (CCD). The optimum condition is obtained when the variables are set to: pH=4, amount of MNPs=20mg, sample volume=30ml, sonication time=1min at room temperature, and desorbing solvent=500µL of methanol. Under the mentioned condition, an extraction recovery of 98.2%, with relative standard deviation of 3.7%, was obtained via five replicated measurements on a 0.1mgL(-1) Brb standard solution. Finally, the method was successfully applied for determination of Brb in human plasma and urine samples. Regarding the method calibration graphs in plasma and urine matrices, spiked in the range of 0.01-200.00µgL(-1), pre-concentration factors of 134 and 122, method detection limit (MDL) of 0.17 and 0.19µgL(-1), and lower limit of quantification of 0.57 and 0.63µgL(-1) were obtained for plasma and urine samples, respectively. The method exhibits good potentials to be a fast, efficient and reliable method for determination of Brb in human biological samples.


Beta-adrenergic responsiveness in cultured aorta smooth muscle cells. Effects of subculture and aging.
beta-Adrenoceptor-mediated vasorelaxation is diminished in vessels from a variety of aged species including humans. This phenomenon was studied for the first time in cultured aorta smooth muscle cells (ASMC) from young (4- to 6-month) and old (24- to 26-month) F-344 rats. Cyclic AMP (cAMP) accumulation was assessed following isoproterenol and forskolin stimulations in primary cultures and after 1-4 passages of aorta smooth muscle cells. Isoproterenol and forskolin increased cAMP accumulation 6- and 10-fold, respectively, in primary cultures from young rats. Isoproterenol stimulation was reduced markedly in passaged cells. Forskolin stimulation was unaffected, indicating passage-related phenotypic changes in receptor-mediated stimulation, but not in post-receptor adenylate cyclase activation. The response to isoproterenol was diminished in old animals, but that to forskolin was unaltered. Thus, cultured ASMC from F-344 rats are highly responsive to beta-adrenoceptor stimulation and demonstrate age-related changes, but undergo phenotypic modulation during passage.


High-throughput analysis of vitamin C in human plasma with the use of HPLC with monolithic column and UV-detection.
Vitamin C plays a central role in the body. One of its important functions is its role as an antioxidant, and accurate measurements are important for interpretations of this role. However, its reactive nature and instability complicates the assessment, especially in biological samples. A high-throughput chromatographic method using monolithic column and UV-detection was developed for the assessment of plasma ascorbic acid and total ascorbic acid. The method showed excellent analytical sensitivity, specificity, precision, recovery and linearity during the validation study. The method was used for the assessment of ascorbic acid and total ascorbic acid during several clinical studies.


The micro-foundations of global business incubation: Stakeholder engagement and strategic entrepreneurial partnerships.
This paper explores the micro-foundations of global business incubation by examining how stakeholders influence the development of technology business incubation platforms and entrepreneurial activities in the context of strategic entrepreneurial partnerships. By drawing from stakeholder theory, we propose a conceptual framework that articulates how stakeholder engagement-including local governments, universities, and business support organizations-varies over time by engaging with entrepreneurs in the form of collaborative partnerships. This paper investigates two technology business incubators in China and the UK through a comparative lens and offers empirical evidence to support its theoretical argumentation. From a temporal perspective, we found that different types of stakeholders play different roles in deploying three distinctive mechanisms-namely, initiating, orchestrating and partaking-during the developmental trajectory involved in the launch and development of entrepreneurial ventures. Importantly, from a micro-foundational perspective, we highlight the agency-orientated activities championed by entrepreneurial organizations situated in global strategic partnership contexts.


Percutaneous coronary intervention in a patient with acute ST-elevation myocardial infarction and hemophilia A.
We report the first case of myocardial infarction in a hemophilia patient which was not directly precipitated by infusion of a clotting factor, and the second hemophilia patient with infarction treated with primary coronary artery stenting. Anticoagulation and platelet inhibition are problematic in such patients. At 2 years after infarction, our patient has experienced neither ischemic nor bleeding complications.


Arthroscopic reconstruction of the medial patellofemoral ligament in skeletally immature patients using the modified sling procedure: a novel technique for MPFL reconstruction.
Patellar dislocation is common in young people. Although isolated anatomic double-bundle reconstruction of the MPFL is a common and effective surgical treatment for patellofemoral instability, concerns about the risk of injury to the epiphysis remain. A total of 21 children and adolescents (9 males, 12 females; mean age: 10.7 years; range: 8 to 13 years) with recurrent patella dislocation or symptomatic instability following a primary dislocation were enrolled in the study. In all patients, double-bundle medial patellofemoral ligament (MPFL) reconstruction and femoral sling procedure were performed under arthroscopy, using an anterior half peroneus longus tendon (AHPLT) autograft. Functional outcomes were evaluated preoperatively and during follow-ups based on Kujala and Lysholm scores. Radiological examinations including radiographs, 3D-CT, and MRI were performed pre- and post-operatively. Among two-year postoperative follow-up (range: 24-42 months) showed significant improvement in functional scores (p < 0.01). The Lysholm score increased from 68 (44.5) to 100 (0) and the Kujala score increased from 26 (34.5) to 100 (2) The patellar tilt angel improved significantly (p < 0.01) from 24.3° ± 10.4 preoperatively to 11.9° ± 7.0 postoperatively. MRIs performed 6- and 12-months post operation did not show any signs of dysfunction of the reconstructed MPFL or cartilage degeneration. Case Series; Level of evidence, 4. Arthroscopic reconstruction of the MPFL using the modified sling procedure is an effective procedure for the treatment of patellar instability in skeletally immature patients.


Cytokinetic differences in the action of N-[2-(dimethylamino)ethyl]acridine-4-carboxamide as compared with that of amsacrine and doxorubicin.
N-[2-(Dimethylamino)ethyl]acridine-4-carboxamide dihydrochloride (DACA) is a topoisomerase II-directed DNA intercalator with high experimental solid-tumour activity. The effect of DACA on the cytokinetics of cultured Lewis lung adenocarcinoma cells was compared with those of two clinical drugs of this class, doxorubicin and amsacrine. Cells were exposed to drugs for a 1-h period at concentrations that reduced viability by approximately 99% as measured by clonogenic assays. Subsequent progress through the cell cycle was monitored by propidium staining of fixed cells and flow cytometry. DACA, amsacrine and doxorubicin did not inhibit the G1- to S-phase transition but did delay progression through the S-phase. The effect was maximal in the late S-phase and, because of the differential rates of progress of cells in various cycle positions, led to the development of a synchronous S-phase peak. This peak moved to the G2/M-phase position at 11 h after the removal of DACA or at 14 h after the removal of amsacrine or doxorubicin. The effects of the drugs on cells initially in the G2-phase was measured by scoring mitotic cells in the presence and absence of colchicine. DACA had an immediate inhibitory effect on the progression of cells from the G2-phase to mitosis. This effect was much greater for DACA than for the other two drugs, consistent with the greater effect of DACA on the G2/M-phase to G1-phase transition. The results suggest that DACA causes cell-cycle changes expected for a DNA-damaging drug but differs from doxorubicin and amsacrine mainly by its effect on the transition of G2-phase cells to mitosis and the G1-phase.


Molecular analysis of constitutive IL-1alpha gene expression in human melanoma cells: autocrine stimulation through NF-kappaB activation by endogenous IL-1alpha.
Constitutive production of/and acquired resistance to anti-proliferative cytokines are implicated in pathogenesis and progression of human melanoma cells. Human melanoma cells A375-C6 are sensitive to interleukin 1 (IL-1) anti-proliferative effect and do not produce IL-1. After long period of culture we have obtained cells which acquired resistance to IL-1. The resistant cells exhibited constitutive production of IL-1α. To analyse the mechanisms that lead to the expression of IL-1α in the cells, we transfected of the resistant clone A375-R8 with CAT (chloramphenicol acetyltransferase) expression plasmids linked to a 5'-flanking deletion mutants of the human IL-1α gene. Two nucleotide regions (--103 to --70 bp) and (--70 to --47 bp) from the start of the first exon appeared to contain a positive regulatory element(s) while the one --421 to --103 bp contained a negative regulatory element(s).The --103 to --70 bp region contained the consensus NF(-k)B (nuclear factor-kB) binding motif.(Immunofluorescence analysis revealed that NF-kB is activated in A375-R8 cells. IL-1 receptor antagonist (IL-1Ra) decreased the level of IL-1α mRNA and production of IL-1α. IL-1Ra also inhibited the localization of p65 in the nuclei and CAT activity in transfectants with the plasmids containing NF-kB binding motif. These results indicate that endogenous IL-1α stimulates the gene expression and production of IL-1α in an autocrine manner through activation of NF-kB.


The Procrustean bed of EU food safety notifications via the Rapid Alert System for Food and Feed: does one size fit all?
Previous reports demonstrated wide variations in contributions by EU Member States (MS) to the Rapid Alert System for Food and Feed (RASFF) especially for border notifications, emphasising that MS with major entry points play a vital role in ensuring that EU food imports meet EU standards. To further explore the variation this paper aimed to examine notification practices among MS by comparing the levels of detection as a function of the total food imported and population. RASFF notifications issued between 2003 and 2007 were analysed using descriptive statistics and network analysis for differences in notification practice between MS. Major variations in contributions to the RASFF database were observed, which did not correlate with MS size or population. For the key contrast of ratio: 'border: non-border notifications', variations between 7%:89% were observed for the average monthly contributions and, import tonnage per border notification revealed up to 129-fold differences between MS. In conclusion, wide variations in food safety practice exist between MS, including both number and type of contributions to the RASFF database, with some MS being relatively highly active in the key class of border notifications. These findings should inform EU food safety enforcement policies and practices; and central resource allocations.


Interactions among mitochondrial proteins altered in glioblastoma.
Mitochondrial dysfunction is putatively central to glioblastoma (GBM) pathophysiology but there has been no systematic analysis in GBM of the proteins which are integral to mitochondrial function. Alterations in proteins in mitochondrial enriched fractions from patients with GBM were defined with label-free liquid chromatography mass spectrometry. 256 mitochondrially-associated proteins were identified in mitochondrial enriched fractions and 117 of these mitochondrial proteins were markedly (fold-change ≥ 2) and significantly altered in GBM (p ≤ 0.05). Proteins associated with oxidative damage (including catalase, superoxide dismutase 2, peroxiredoxin 1 and peroxiredoxin 4) were increased in GBM. Protein-protein interaction analysis highlighted a reduction in multiple proteins coupled to energy metabolism (in particular respiratory chain proteins, including 23 complex-I proteins). Qualitative ultrastructural analysis in GBM with electron microscopy showed a notably higher prevalence of mitochondria with cristolysis in GBM. This study highlights the complex mitochondrial proteomic adjustments which occur in GBM pathophysiology.


Visualization and quantification of neurokinin-1 (NK1) receptors in the human brain.
This study was conducted to develop a new positron emission tomography (PET) method to visualize neurokinin-1 (NK(1)) receptor systems in the human brain in vivo in order to examine their neuroanatomical distribution and facilitate investigations of the role of substance P, NK(1) receptors, and NK(1) receptor antagonists in central nervous system (CNS) function and dysfunction. PET studies were conducted in 10 healthy male volunteers using a novel selective, high-affinity NK(1) receptor antagonist labeled with fluorine-18 to very high specific radioactivity (up to 2000 GBq/micromol) [F-18]SPA-RQ. Data were collected in 3D mode for greatest sensitivity. Different modeling methods were compared and regional receptor distributions determined for comparison with in vitro autoradiographic studies using postmortem human brain slices with [F-18]SPA-RQ. The studies showed that the highest uptake of [F-18]SPA-RQ was observed in the caudate and putamen. Lower binding was found in globus pallidus and substantia nigra. [F-18]SPA-RQ uptake was also widespread throughout the neocortex and limbic cortex including amygdala and hippocampus. There was very low specific uptake of the tracer in the cerebellar cortex. The distribution pattern was confirmed using in vitro receptor autoradiography with [F-18]SPA-RQ on postmortem human brain slices. Kinetic modeling of the [F-18]SPA-RQ uptake data indicated a binding potential between 4 and 5 in the basal ganglia and between 1.5 and 2.5 in the cortical regions. [F-18]SPA-RQ is a novel tool for exploration of the functions of NK(1) receptors in man. [F-18]SPA-RQ can be used to define receptor pharmacodynamics and focus dose selection of novel NK(1) receptor antagonists in clinical trials thereby ensuring adequate proof of concept testing particularly in therapeutic applications related to CNS dysfunction.


Descending pathways modulating the spinal circuitry for ejaculation: effects of chronic spinal cord injury.
Sexual dysfunction is a common complication in men with chronic spinal cord injury. In particular, ejaculation is severely compromised or absent and the resulting infertility issues are important to this group of predominantly young men. To investigate the neural circuits and descending spinal pathways involved in ejaculation, animal models have been developed in normal and spinal cord-injured preparations. Primarily through studies in rats, spinal ejaculatory circuits have been described including (i) autonomic circuits at the thoracolumbar and lumbosacral levels mediating the emission phase of ejaculation, (ii) somatic circuits at the lumbosacral level controlling the expulsion phase of ejaculation through sequential and rhythmic contraction of perineal striated muscles (e.g. bulbospongiosus), and (iii) a proposed ejaculatory pattern generator in the lumbar cord. Midthoracic incomplete chronic spinal cord injury has revealed the dependency of spinal ejaculatory circuits on bilateral spinal pathways from the brainstem via modulation of pudendal motor neuron reflexes and pudendal nerve autonomic fibers. Accordingly, sensory input from the dorsal nerve of the penis, required to trigger the ejaculatory response in animals and humans, is no longer inhibited from the lateral paragigantocellularis nucleus in the ventrolateral medulla. This inhibitory effect, likely presynaptic through a serotonergic pathway, is thought to be necessary to provide the rhythmic, bursting, and sequential contractions of the perineal muscles during ejaculation. Chronic lateral hemisection injury, which severs half of the descending lateral funiculus-located pathways, results in new functional connections of the pudendal reflex inhibitory and pudendal sympathetic activation pathways across the midline, above and below the lesion, respectively. Clinical correlations in spinal cord-injured men have demonstrated the validity of the rodent animal for the study of ejaculatory dysfunction after chronic injury.


[Morphology of the cranial base in a group of Campania school children].
The A.A. analysed the cranial base morphology on 14 school-children in the eastern part of Naples, to establish if the most used cephalometric values were useful for Campanian people. The values found out resulted superimposed to those referred in the considered analysis.


Overlapping prescriptions of opioids, benzodiazepines, and carisoprodol: "Holy Trinity" prescribing in the state of Florida.
High-risk combinations of controlled medications, such as those involving opioid analgesics, are under increased scrutiny because of their contribution to the opioid epidemic in the United States. Responsible prescribing guidelines indicate that the triple drug combination--opioids, benzodiazepines and skeletal muscle relaxants, especially carisoprodol--should not be concurrently prescribed. This pharmacoepidemiologic study was designed to primarily examine the characteristics of patients receiving this triple combination compared to the group receiving only opioids and benzodiazepines. Results show that, while the number of exposed patients has declined since 2012, approximately 17,000 Floridians were prescribed this combination in 2017 alone. Demographically, recipients of these prescriptions were younger, more likely to be female, and geographically-localized. Furthermore, these patients were more frequently associated with a prescriber in the top 1% of opioid and/or benzodiazepine prescribing, have more multiple provider episodes ("doctor shopping"), and receive higher mean daily opioid dosages. These findings raise important questions as to how frequently prescribers are checking prescription drug monitoring programs, following US Centers for Disease Control and Prevention opioid prescribing guidelines, and/or handling the clinical challenges associated with pharmaceutical management of patients with complex, painful health conditions.


The role of iodine-123-labeled 15-(p-iodophenyl)-3R,S-methylpentadecanoic acid scintigraphy in the detection of local myocardial involvement of sarcoidosis.
The purpose of this study is to examine the role of Iodine-123-labeled 15-(p-iodophenyl)-3R,S-methylpentadecanoic acid (BMIPP) scintigraphy in patients with cardiac sarcoidosis. Study materials were six patients with pathologically proven cardiac sarcoidosis. BMIPP and resting Thallium-201 (201Tl) myocardial scintigraphy, echocardiography, were performed within 22 days in each patient. Myocardium was divided into nine areas per one case. A total of 24 areas had involvement by sarcoidosis. A total of 18 areas with defects of BMIPP accumulation and 14 areas with defects of 201Tl accumulation were detected. A total of 12 areas were determined as showing reduced wall motion. The sensitivity, specificity, positive and negative predictive values of wall motion abnormality for the detection of myocardial involvement were 50%, 100%, 100% and 71%. The sensitivities of BMIPP and 201Tl scintigraphy for the detection of local myocardial involvement were 75% and 58%. The specificities of BMIPP and 201Tl scintigraphy were both 100%. The positive predictive values of BMIPP and 201Tl scintigraphy were both 100%. The negative predictive values of BMIPP and 201Tl scintigraphy were 83% and 75%. BMIPP scintigraphy was more sensitive and had a higher negative predictive value compared to 201Tl scintigraphy and echocardiography for the detection of myocardial involvement of sarcoidosis.


A Systematic Review of the Effectiveness of Interventions to Promote Referral; Adherence; and Uptake of Pulmonary Rehabilitation for Patients with Chronic Obstructive Pulmonary Disease.
Pulmonary rehabilitation (PR) is an effective treatment for patients with chronic obstructive pulmonary disease (COPD). However, referral, uptake, and adherence remain low. To determine effectiveness of interventions to increase patient referral, uptake, and adherence to PR programs for patients with COPD. Randomized controlled trials (RCTs), non-randomized controlled trials, pre-post studies, and uncontrolled studies were sought from 7 databases and 3 clinical trial registries, to end August 2021. Full articles/conference abstracts were included if a coordinated set of activities was targeted to healthcare professionals (HCPs) caring for COPD patients, adults with COPD or their carers, to increase referral, uptake or adherence to any type of PR program. Two review authors independently screened titles, abstracts and full texts, extracted data and critically appraised studies using standard risk of bias tools. From 11,272 records, 30 studies (23 full-text; 7 abstracts) met inclusion criteria: study interventions and designs were varied and generally low quality, targeting patients (n=13), HCPs (n=14) or both (n=3 studies). A CCT of patient held evidence score cards increased referral by 7.3% compared to 1.3% for usual care (p-0.03). A cluster RCT involving COPD nurse home visits with individualized care plans increased uptake to 31% compared to 10% in usual care (p=0.002). For people with anxiety or depression, one RCT of cognitive behavioral therapy alongside PR increased adherence (mean sessions 14.0 (sd 1.7) compared to 12.4 (sd 2.6)). Although a small number of studies, the weight of evidence suggested that interventions incorporating partnership working between patients and HCPs appeared to increase referral, uptake, and adherence with greater effectiveness than those targeting single populations. Increasing knowledge and empowering HCPs and patients may be important strategies. Concerns about study design and risk of bias suggest clear need for well-designed trials of interventions to report full pathway outcomes.


Dose and time related changes in LDH-X activity, epididymal carnitine levels and fertility, in gossypol-treated male rats.
Gossypol acetic acid could induce total infertility in male Wistar/NIN strain rats at a dose of 30 mg per kg body weight per day, for 52 days. 20 mg for 52 days and 30 mg for 38 days produced partial infertility. Highly significant correlation between parameters of infertility like reduced cauda sperm count, and implantation sites, and reduction in LDH-X activity of cauda epididymal sperm and carnitine levels in cauda epididymal fluid were observed. Testis remained unaffected.


Investigating the sense of agency and its relation to subclinical traits using a novel task.
Tasks measuring the sense of agency often manipulate the predictability of action outcomes by introducing spatial deviation. However, the extent to which spatial predictability of an outcome influences the sense of agency when spatial deviation is controlled for remains untested. We used a novel task to investigate the effect of several factors (action-outcome contingency, spatial deviation, and spatial predictability when controlling for spatial deviation of action outcomes) on the sense of agency. We also investigated trait predictors of metacognition of agency-the degree to which participants' confidence in their agency judgements corresponds to the accuracy of those judgements. Initial and replication samples completed contingency, deviation, and predictability versions of the task. Across samples, participants' sense of agency was impacted by action-outcome contingency and spatial deviation of action outcomes. Manipulation of the spatial predictability of action outcomes did not reliably impact the sense of agency. Metacognition of agency was related to alexithymic traits-higher alexithymia scores were associated with reduced metacognition of agency.


Quality of registration of maternal deaths in Mozambique: a community-based study in rural and urban areas.
Maternal mortality is an important indicator used to assess the level of development of a country and particularly the coverage of health services. In Africa where the maternal mortality ratios are about 100 times higher than those of developed countries the situation is even worse due to problems of underregistration. In Mozambique the figures officially used are derived from the data of some hospitals in the main cities. This leaves out the 60% of pregnant women who do not attend health facilities. It was against this background that the present study was designed to be a prospective community-based study carried out in the central region of Mozambique involving a population of 207,369 inhabitants. The study covered a 12-month period, from 01/08/96 to 31/07/97 and found 214 deaths among women in reproductive age (15-49 yr) of whom 40 were classified as maternal deaths. Gross underregistration was revealed, as the health institutions failed by 86% in recording maternal deaths, the same happening with other official sources, like the Civil Register and the Funeral Services registering only 9% of the maternal deaths. This kind of study is however complex and labour intensive and a practical way of monitoring has to be found; for the region where the study took place close collaboration with private cemeteries appears to be a way to be further explored.


Postnatal expression of opioid genes in rat brain.
Developmentally regulated expression of opioid genes in rat brain was studied by Northern blot hybridization, using cloned rat genes as probes. Total RNA was isolated from the rat medulla oblongata, hypothalamus and striatum at 11 stages of development from birth to postnatal day 28. The steady-state levels of opioid precursor mRNAs in the above tissues were determined by hybridization to 32P-labeled opioid DNA probes and densitometric scanning of the appropriate bands. Analysis of the relative steady-state concentrations of opioid mRNAs has shown that the tissue distribution and size of these mRNAs during postnatal development of the CNS are similar to the pattern described in the adult. However, up to 50-fold changes in the relative mRNA concentrations were observed. These changes are tissue-specific, characterized by a peak within the postnatal stage of development, and are followed by a build-up of mRNA levels characteristic of the adult. Moreover, comparison of the changes in mRNA levels of proenkephalin to those of prodynorphin at their main site of synthesis in the brain, namely the striatum, revealed striking similarities in pattern. The association of this phenomenon with a possible role of opioid genes in development is suggested.


Advanced statistics identification of participant and treatment predictors associated with severe adverse effects induced by fluoropyrimidine-based chemotherapy.
Adverse effects following fluoropyrimidine-based chemotherapy regimens are common. However, there are no current accepted diagnostic markers for prediction prior to treatment, and the underlying mechanisms remain unclear. This study aimed to determine genetic and non-genetic predictors of adverse effects. Genomic DNA was analyzed for 25 single nucleotide polymorphisms (SNPs). Demographics, comorbidities, cancer and fluoropyrimidine-based chemotherapy regimen types, and adverse effect data were obtained from clinical records for 155 Australian White participants. Associations were determined by bivariate analysis, logistic regression modeling and Bayesian network analysis. Twelve different adverse effects were observed in the participants, the most common severe adverse effect was diarrhea (12.9%). Bivariate analysis revealed associations between all adverse effects except neutropenia, between genetic and non-genetic predictors, and between 8 genetic and 12 non-genetic predictors with more than 1 adverse effect. Logistic regression modeling of adverse effects revealed a greater/sole role for six genetic predictors in overall gastrointestinal toxicity, nausea and/or vomiting, constipation, and neutropenia, and for nine non-genetic predictors in diarrhea, mucositis, neuropathy, generalized pain, hand-foot syndrome, skin toxicity, cardiotoxicity and fatigue. The Bayesian network analysis revealed less directly associated predictors (one genetic and six non-genetic) with adverse effects and confirmed associations between six adverse effects, eight genetic predictors and nine non-genetic predictors. This study is the first to link both genetic and non-genetic predictors with adverse effects following fluoropyrimidine-based chemotherapy. Collectively, we report a wealth of information that warrants further investigation to elucidate the clinical significance, especially associations with genetic predictors and adverse effects.


Systemic adjuvant therapies and radiotherapy to the conserved breast: strategies revisited.
The rapid increase in the use of both radiotherapy and systemic therapy for patients with early breast cancer requires reconsideration of the strategies that have been used in the past for combining both treatments. In these considerations the impact of both radiotherapy as well as systemic therapy on local control and survival of breast cancer have to be considered. An important point for discussion has often been the sequencing of radiotherapy and systemic treatment. Nowadays the possibilities of trastuzumab for her-2 neu positive breast cancer leads to the question whether or not this can be combined with radiotherapy. Finally new possibilities of sophisticated radiotherapy approaches allow further minimizing the late toxicity of radiotherapy.


Runoff mitigation via micro-dams and conservation tillage-Numerical modeling of runoff and erosion from maize field trials.
Runoff and erosion are the most important transport pathways of water, sediment, and associated pesticides from sloped agricultural fields. This results in the loss of fertile topsoil material, nutrients, irrigation water, and plant protection products (PPP) into adjacent surface water bodies. In the European and US risk assessment for the registration of PPP, runoff and erosion are numerically calculated with the simulation Pesticide Root Zone Model (PRZM) using the US Department of Agriculture (USDA) runoff curve number (CN) concept for the water movement and the MUSS equation to quantify the sediment transfer. This work presents an evaluation of maize field trials conducted in three seasons that considered micro-dams (i.e., small earthen dams between the rows; also known as "furrow diking," "furrow damming," etc.) and/or conservation tillage (via subsoiling) as mitigation measures to investigate the effects on the reduction in runoff and erosion. Measured quantitative reductions and event-wise calculated CN are presented. Furthermore, the trials were simulated using the PRZM over the complete vegetation period and runoff CN as well as parameter values of the MUSS erosion equation (a relative adaptation of the C-factor) were inversely estimated. Compared with the control plots (i.e., conventional tillage), micro-dams or conservation tillage reduced runoff by 24%-71% or 69%-89%, and erosion by 54%-81% or 91%-98%. Based on these data, a robust case can be made to lower CN or parameters in the MUSS equation for surface water exposure scenarios to consider the effects on predicted environmental concentrations (PECs) and estimated environmental concentrations (EECs). Mean resulting CN reductions by micro-dams or conservation tillage were ascertained to be 6% (±2.5%) or 12% (±3.0%), the C-factor was reduced by a factor of 0.1 (±0.15) or 0.48 (±0.19). Example calculations show reductions in the ranges of 11%-100% for PECs and 30%-98% for EECs. Integr Environ Assess Manag 2022;18:1348-1363. © 2021 Bayer AG Crop Science. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).


Controlled Intracellular Polymerization for Cancer Treatment.
Numerous prodrugs have been developed and used for cancer treatments to reduce side effects and promote efficacy. In this work, we have developed a new photoactivatable prodrug system based on intracellular photoinduced electron transfer-reversible addition-fragmentation chain-transfer (PET-RAFT) polymerization. This unique polymerization process provided a platform for the synthesis of structure-predictable polymers with well-defined structures in living cells. The intracellularly generated poly(N,N-dimethylacrylamide)s were found to induce cell cycle arrest, apoptosis, and necroptosis, inhibit cell proliferation, and reduce cancer cell motilities. This polymerization-based "prodrug" system efficiently inhibits tumor growth and metastasis both in vitro and in vivo and will promote the development of targeted and directed cancer chemotherapy.


Development of an indirect competitive ELISA method based on ORF2 detecting the antibodies of novel goose astrovirus.
Goose astrovirus (GAstV) characterized by articular and visceral gout, is an emerging pathogen with a wide distribution on mainland China, leading to serious economic losses in the goose-raising industry. Because vaccines to prevent GAstV infections are not available currently, early diagnosis is critical when treating symptomatic geese and in preventing GAstV transmission. In this context, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) to detect GAstV-specific antibodies from geese was developed, and the detections were optimized. A series of experiments proved that the ic-ELISA shows excellent diagnostic performance and discriminatory power with high sensitivity and specificity. The ic-ELISA for GAstV detection was applied on 67 field serum samples, and comparing the detection results with the virus neutralization test verified the accuracy of the ic-ELISA. The correlation coefficient between the ic-ELISA and the virus neutralization test was 80%, demonstrating the proposed ic-ELISA method could be a useful and effective tool for the diagnostic, serological epidemiological investigation and immune monitoring of the GAstV in goose-producing regions.


Genotoxic and morpho-physiological responses of ZnO macro- and nano-forms in plants.
In the current study, two plants, viz., Pisum sativum L. and Hordeum vulgare L., were exposed to nano- and macro-dispersed ZnO at 1, 10, and 30 times of maximal permissible concentration (MPC). The main objective of the study is to depict and compare the genotoxicity in terms of chromosomal anomalies, cytotoxicity (i.e., mitotic index), and phytotoxicity (viz., germination, morphometry, maximal quantum yield, and chlorophyll fluorescence imaging) of macro- and nano-forms of ZnO along with their accumulation and translocation. In the case of genotoxic and cytotoxic responses, the maximal effect was observed at 30 MPC, regardless of the macro- or nano-forms of ZnO. The phytotoxic observations revealed that the treatment with macro- and nano-forms of ZnO significantly affected the germination rate, germination energy, and length of roots and shoots of H. vulgare in a dose-dependent manner. The factor toxicity index of treated soil demonstrated that toxicity soared as concentrations increased and that at 30 MPC, toxicity was average and high in macro- and nano-dispersed ZnO, respectively. Furthermore, the photosynthetic parameters were observed to be negatively affected in both treatments, but the maximal effect was observed in the case of nano-dispersed form. It was noted that the mobility of nano-dispersed ZnO in the soil was higher than macro-dispersed. The increased mobility of nano-dispersed ZnO might have boosted their accumulation and translocation that subsequently led to the oxidative stress due to the accelerated production of reactive oxygen species, thus strengthen toxicity implications in plants.


Are there significant gradients of pO2 in cells?
It is widely recognized that the intracellular oxygen tension (pO2) plays an important role in cellular function and metabolism. In experimental and theoretical consideration involving the role of the pO2 the values that are used usually are those for the pO2 at the exterior of cells, because these values can be more readily measured. Such an approach is based on the assumption that the intracellular pO2 is very similar to the extracellular pO2 because oxygen freely diffuses across cell membranes and within cells, at a rate that is similar to that in the extracellular media. For the past several years we have been developing and applying electron paramagnetic resonance (EPR) techniques to test directly whether there are intracellular gradients of pO2 and if so, where they occur and what factors determine them. We previously reported significant gradients between the average pO2 in the intracellular and extracellular compartments in cell suspensions (Glockner 1989). More recently we developed techniques that enabled us to measure simultaneously the concentration of oxygen within a specific compartment, the phagosomes of activated macrophages, and the extracellular compartment and found gradients of up to 48 microM under some conditions (James 1995). The precise mechanism of the intracellular-extracellular oxygen gradient remains uncertain. The possibilities include that the diffusion of oxygen is not as free as assumed (e.g. that the cell membrane can act as a barrier) and the occurrence of active transport of O2 out of the cells. We report here on the use of a simple theoretical approach to evaluate the values of three key parameters which might account for the observed intracellular-extracellular oxygen measurements: (1) oxygen consumption; (2) the diffusion coefficient of oxygen in the cytosol; (3) the solubility of oxygen in the cytosol. We used two different models for the relationship between the oxygen consuming compartment (assumed to be primarily the mitochondria) and the intracellular compartment in which the measurements were made (especially phagosomes): uniform and non-uniform distribution of the mitochondria. Using these models and consensus values from the literature, we were unable to account for the experimentally observed differences in pO2 between the intracellular and extracellular compartments. Also we found that with the variation of any one parameter we could not plausibly account for the measurements made in the phagosomal and extracellular compartments. There are at least three logical possibilities to account for these results: 1) this methodology is erroneous and/or produces artifacts in the system resulting in invalid results; 2) the observation of a gradient in oxygen concentration between these two compartments arises from significant simultaneous variations of more than one of the critical parameters which are used conventionally to calculate potential gradients in pO2; 3) there is another factor not considered in the model which accounts for the observation (e.g. active transport; significantly higher than expected barriers to oxygen diffusion in the membrane).


Green solvents and Ultrasound-Assisted Extraction of bioactive orange (Citrus sinensis) peel compounds.
Byproducts such as orange peel have potential uses because of their bioactive compounds, which are important for their potential to reduce the risk factors of diseases caused by aging. The lack of effective techniques and the high levels of pollution produced by the conventional extraction of bioactive compounds using organic solvents have highlighted the need to enhance the 'green chemistry' trend. This study evaluates the use of ultrasound to extract bioactive compounds from orange peel. The antioxidant capacity, phenolic content, ascorbic acid, total carotenoids, and HPLC profile of phenolic compounds from orange peel extracts were obtained by a physicochemical evaluation. The results demonstrate that the optimal conditions for the ultrasound-assisted extraction of bioactive orange peel compounds were a power of 400 W, a time of 30 min, and 50% ethanol in water. These conditions were used to obtain a total carotenoid concentration of 0.63 mg ß-carotene/100 g, vitamin C concentration of 53.78 mg AA/100 g, phenolic concentration of 105.96 mg GAE/100 g, and antioxidant capacity of ORAC = 27.08 mM TE and TEAC = 3.97 mM TE. The major phenolic compound identified in all orange peel extracts was hesperidin, with a maximum concentration of 113.03 ± 0.08 mg/100 g.


Determination of B-trichothecenes in wheat by post column derivatisation liquid chromatography with fluorescence detection (PCD-HPLC-FLD).
B-trichothecenes are one of the most common contaminants of cereals in Europe. Therefore, the use of fast and accurate methods is necessary to measure contamination levels and observe regulatory limits.At the moment, mostly gas chromatographic (GC) methods are used but HPLC-UV methods are also employed. Clean-up is commonly done either with immunoaffinity or Mycosep® columns.In the Christian Doppler Laboratory for Mycotoxin Research we have established an alternative HPLC method with post column derivatisation (PCD) as an alternative to existing chromatographic methods. This PCD-HPLC-FLD method uses a Mycosep® clean-up and allows the simultaneous detection and quantification of deoxynivalenol, nivalenol, fusarenon X, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol. A validation with wheat gave for deoxynivalenol a limit of quantification ten times below the drafted European Union guideline level (500 µg.kg(-1)) and a limit of detection of 8 µg.kg(-1). The relative standard derivation for DON was 10% (n=30). The obtained mean recovery rate for DON was 90% in a range from 50 to 1000 µg.kg(-1).


Variant Two-Stage Ileal Pouch-Anal Anastomosis: An Innovative and Effective Alternative to Standard Resection in Ulcerative Colitis.
Ulcerative colitis patients have been historically treated with standard single, 2-, and 3-stage operative approaches. We perform a variant 2-stage procedure beginning with total abdominal colectomy and end ileostomy followed by completion proctectomy and ileal pouch-anal anastomosis (IPAA) without a diverting loop ileostomy. This study evaluates the effectiveness of this innovative alternative. Patients with ulcerative colitis, admitted to the University of North Carolina Hospital between 2003 and 2010 for IPAA, were eligible for inclusion. The 3-year cumulative incidence of pouch leaks among patients undergoing variant 2-stage were compared with those undergoing classic 2-stage, using inverse probability-of-treatment weighted Kaplan- Meier survival curves, and 95% CIs were estimated using nonparametric bootstrapping. There were 248 patients who underwent IPAA; 139 (56.1%) underwent classic 2-stage and 109 (43.9%) underwent variant 2-stage. After standardization, there was no significant difference in the 3-year cumulative incidence of pouch leaks between patients undergoing variant 2-stage, compared with the standard single- or 2-stage procedure (risk difference 0.01; 95% CI -0.08, 0.15). At the time of the first surgical procedure, patients undergoing a variant 2-stage were more likely to have lower BMIs (median 22.5 kg/m2 vs 26.7 kg/m2; p < 0.0001), an urgent/emergent procedure (56.9% vs 0.0%; p < 0.0001), biologic use within 2 weeks of surgery (32.1% vs 17.5%; p = 0.003), and high dose steroid use (60.4% vs 16.7%; p ≤ 0.0001). Variant 2-stage IPAA is a safe and effective operative approach with comparable outcomes in a more acute population based on BMI, steroid use, and urgency of operation.


The effect of DSM-5 criteria on externalizing, internalizing, behavioral and adaptive symptoms in children diagnosed with autism.
Diagnostic criteria for autism spectrum disorders (ASDs) are changing with the fifth edition of the Diagnostic and Statistical Manual (DSM-5), which simplifies the diagnostic categories into social/emotional deficits and repetitive and restricted behavior. ASDs have been closely linked to a variety of other disorders, in particular externalizing disorders such as ADHD, and internalizing disorders including anxiety disorders and obsessive compulsive disorder. The present study examines the externalizing, internalizing, behavioral and adaptive symptoms of children with ASD. Children diagnosed with the DSM-IV who do not meet diagnostic criteria for DSM-5 and were compared to a non-ASD sample and a sample of those who meet the new criteria. Differences were examined between the three experimental groups with respect to internalizing, externalizing, behavioral severity and adaptive behavior. No significant differences were observed between the DSM-5 and DSM-IV groups with respect to composite and subscale scores on the externalizing, behavior severity index and adaptive behavior domains of the Behavior Assessment System for Children, Second Edition. Significantly more impairment was evident for both ASD groups compared to the no-ASD group.


Clinical evaluation of intravenous alone versus intravenous or intraosseous access for treatment of out-of-hospital cardiac arrest.
Obtaining vascular access during out-of-hospital cardiac arrest (OHCA) is challenging. The aim of this study was to determine if using intraosseous (IO) access when intravenous (IV) access fails improves outcomes. This was a prospective, parallel-group, cluster-randomised study that compared 'IV only' against 'IV + IO' in OHCA patients, where if 2 IV attempts failed or took more than 90 s, paramedics had 2 further attempts of IO. Primary outcome was any return of spontaneous circulation (ROSC). Secondary outcomes were insertion success rate, adrenaline administration, time to adrenaline and survival outcome. A total of 1007 patients were included in the analysis. An Intention To Treat analysis showed a significant difference in success rates of obtaining vascular access in the IV + IO arm compared to the IV arm (76.6% vs 61.1% p = 0.001). There were significantly more patients in the IV + IO arm than the IV arm being administered prehospital adrenaline (71.3% vs 55.4% p = 0.001). The IV + IO arm also received adrenaline faster compared to the IV arm in terms of median time from emergency call to adrenaline (23 min vs 25 min p = 0.001). There was no significant difference in ROSC (adjusted OR 0.99 95%CI: 0.75-1.29), survival to discharge or survival with CPC 2 or better in both groups. A Per Protocol analysis also showed there was higher success in obtaining vascular access in the IV + IO arm, but ROSC and survival outcomes were not statistically different. Using IO when IV failed led to a higher rate of vascular access, prehospital adrenaline administration and faster adrenaline administration. However, it was not associated with higher ROSC, survival to discharge, or good neurological outcome.


Quantitatively probing the magnetic behavior of individual nanoparticles by an AC field-modulated magnetic force microscopy.
Despite decades of advances in magnetic imaging, obtaining direct, quantitative information with nanometer scale spatial resolution remains an outstanding challenge. Current approaches, for example, Hall micromagnetometer and nitrogen-vacancy magnetometer, are limited by highly complex experimental apparatus and a dedicated sample preparation process. Here we present a new AC field-modulated magnetic force microscopy (MFM) and report the local and quantitative measurements of the magnetic information of individual magnetic nanoparticles (MNPs), which is one of the most iconic objects of nanomagnetism. This technique provides simultaneously a direct visualization of the magnetization process of the individual MNPs, with spatial resolution and magnetic sensitivity of about 4.8 nm and 1.85 × 10(-20) A m(2), respectively, enabling us to separately estimate the distributions of the dipolar fields and the local switching fields of individual MNPs. Moreover, we demonstrate that quantitative magnetization moment of individual MNPs can be routinely obtained using MFM signals. Therefore, it underscores the power of the AC field-modulated MFM for biological and biomedical applications of MNPs and opens up the possibility for directly and quantitatively probing the weak magnetic stray fields from nanoscale magnetic systems with superior spatial resolution.


Bimatoprost promotes satiety and attenuates body weight gain in rats fed standard or obesity-promoting diets.
Bimatoprost is a synthetic prostamide F2α analog that down-regulates adipogenesis in vitro. This effect has been attributed to participation in a negative feedback loop that regulates anandamide-induced adipogenesis. A follow-on investigation has now been conducted into the broader metabolic effects of bimatoprost using rats under both normal state and obesity-inducing conditions. Chronic bimatoprost administration attenuated weight gain in a dose dependent-manner in rats fed either standard [max effect -7%] or obesity-promoting diets [max effect -23%] over a 9-10 week period. Consistent with these findings, bimatoprost promoted satiety as measured by decreased food intake [max effect, -7%], gastric emptying [max effect, -33-50%] and decreased circulating concentrations of the gut hormones, ghrelin and GLP-1 [max effect, -33-50%]. Additionally, subcutaneous, and visceral fat mass were distinctly affected by treatment [-30% diet independent]. Taken together, these results suggest that bimatoprost regulates energy homeostasis through promoting satiety and a decrease in food intake. These newly reported activities of bimatoprost reveal an additional method of metabolic disease intervention for potential therapeutic exploitation.


Low-flow perfusion preservation versus static preservation for isolated rat heart: effects on recovery of myocardial function.
Clinically, donor hearts cannot be preserved for >6 hours between explantation and recipient implantation. A better approach is needed to preserve donor hearts for a longer time. We tested whether low-flow perfusion (LFP) could satisfactorily preserve isolated rat hearts with histidine-tryptophan-ketoglutarate (HTK) solution or Fuwai modified (FWM) solution. We divided 32 male Sprague-Dawley rats randomly into 4 groups (n = 8): H1, H2, F1, and F2. The Langendorff heart model immersed isolated hearts in the H1 and F1 groups in HTK or FWM solution for 8 hours at 4 °C. Isolated hearts in the H2 and F2 groups were low-flow perfused with HTK solution and FWM solution for 8 hours at a pressure of 10 cmH(2)O at 4 °C. After 60 minutes reperfusion, we measured recovery of cardiac function, myocardial water content, and leakage of myocardial enzymes. After reperfusion, no cardiac rebeating was observed among F1 group hearts; in addition, they showed significantly higher myocardial water content and lactate dehydrogenase leakage compared with the other 3 groups (P < .05). The recovery rates of cardiac function among H2 hearts were better than the other 3 groups (P < .05); their myocardial water content and enzyme leakage were less than the other 3 groups (P < .05). Hypothermic LFP was better than static storage to preserve isolated rat hearts. HTK solution afforded better myocardial protection than FWM.


Macrophage-mediated mitogenic suppression induced in mice of the C3H lineage by a vaccine strain of Salmonella typhimurium.
Salmonella typhimurium, strain SL3235, an avirulent organism, has been used as a live vaccine in mice of the C3H lineage and has been found to confer high levels of protection. In the present study, it was found that intraperitoneal injection of approximately 5 X 10(5) live SL3235 induced potent suppression of spleen cell mitogenic responses to a panel of B- and T-cell mitogens in the Salmonella-hypersusceptible C3H/HeJ and C3HeB/FeJ, and the inherently resistant C3H/HeNCrlBR mice. Maximal suppression (greater than 99%) was seen at 1 week, and was still significant but waning (50%) at 3 weeks postimmunization. In contrast, cells of mice receiving acetone-killed cells were not suppressed. Removal of macrophages, but not T or B cells, restored responsiveness, indicating that suppression was macrophage mediated. Prostaglandins were not the major mediator of suppression, as in vitro administration of indomethacin failed to abrogate suppression. As mitogenic suppression occurred in mice with high levels of Salmonella immunity, the suppression is interpreted as a marker of a powerful immunomodulatory process induced by live cells, rather than as an indication of poor immune status of the host.


Suppression of CCR5-tropic HIV type 1 infection by OX40 stimulation via enhanced production of β-chemokines.
To elucidate the immunological role for the costimulatory molecule OX40 against the early stage of HIV-1 infection, fresh peripheral blood mononuclear cells (PBMCs) from normal donors were stimulated with immobilized anti-CD3 monoclonal antibody (mAb) together with soluble anti-CD28 mAb for 24 h, infected with CCR5-tropic (R5) HIV-1, and then cocultured in the presence or absence of OX40 ligand (OX40L). Results of these studied showed that OX40 stimulation led to a marked reduction in levels of p24, the frequency of intracellular p24(+) cells, as well as HIV-1-mediated syncytium formation. The suppression was reversed by anti-OX40L mAb. The mechanism underlying the R5 HIV-1 suppression was shown to be mediated in part by the CCR5-binding β-chemokines RANTES, MIP-1α, and MIP-1β, since the effect of the OX40 stimulation was reversed by a neutralizing antibody mixture against these three β-chemokines. Thus, OX40 stimulation enhanced the production of these CCR5-binding β-chemokines by the activated PBMCs and subsequently down-modulated CCR5 expression on the activated CD4(+) T cells. Taken together, the present data revealed a new role for OX40 in HIV-1 infection and documents the fact that OX40 stimulation suppresses the infection of primary activated PBMCs with R5 HIV-1 via enhanced production of R5 HIV-1 suppressing β-chemokines.


Control of hepatic triacylglycerol synthesis. Diurnal variations in hepatic phosphatidate phosphohydrolase activity and in the concentrations of circulating insulin and corticosterone in rats.
Male rats were kept for 14 days with alternating 12h periods of light and darkness. The hepatic activity of soluble phosphatidate phosphohydrolase and the concentration of serum insulin were maximum at about 2h after dark. The peak concentration of serum corticosterone occurred 2h before the dark period. It is proposed that corticosterone is partly responsible for the increased phosphohydrolase activity, and that this enables the liver to increase its capacity to synthesize triacylglycerols during the period of maximum feeding.


Exploring the distribution of park availability, features, and quality across Kansas City, Missouri by income and race/ethnicity: an environmental justice investigation.
Parks are key community assets for physical activity, but some evidence suggests these resources are not equitably distributed. This study examined disparities by income and race/ethnicity in the availability, features, and quality of parks across Kansas City, Missouri. All parks and census tracts (CTs) were mapped using geographical information systems, and park features and quality were determined via audits. Multivariate analyses of covariance analyzed differences in park availability, features, and quality across low-, medium-, and high-income and race/ethnicity CT tertiles. Low-income CTs contained significantly more parks, but also had fewer parks with playgrounds and more quality concerns per park. High minority CTs had more parks with basketball courts, but fewer parks with trails. Medium-income CTs contained more aesthetic features per park. Future research should examine policies that contribute to and that might rectify disparities in park features and quality, especially in low-income and high minority areas.


Visual pigment: G-protein-coupled receptor for light signals.
The visual pigment present in photoreceptor cells is a prototypical G-protein-coupled receptor (GPCR) that receives a light signal from the outer environment using a light-absorbing chromophore, 11-cis-retinal. Through cis-trans isomerization of the chromophore, light energy is transduced into chemical free energy, which is in turn utilized for conformational changes in the protein to activate the retinal G-protein. In combination with site-directed mutagenesis, various spectroscopic and biochemical studies identified functional residues responsible for chromophore binding, color regulation, intramolecular signal transduction and G-protein coupling. Extensive studies reveal that these residues are localized into specific domains of visual pigments, suggesting a highly manipulated molecular architecture in visual pigments. In addition to the recent findings on dysfunctional mutations in patients with retinitis pigmentosa or congenital night blindness, the mechanism of intramolecular signal transduction in visual pigments and their evolutionary relationship are discussed.


Using the intraoperative hand held probe without lymphoscintigraphy or using only dye correlates with higher sensory morbidity following sentinel lymph node biopsy in breast cancer: a review of the literature.
There are no studies that have directly investigated the incremental reduction in sensory morbidity that lymphoscintigraphy images (LS) and triangulated body marking or other skin marking techniques provide during sentinel lymph node biopsy (SLNB) compared to using only the probe without LS and skin marking or using only dye. However, an indirect assessment of this potential for additional sensory morbidity reduction is possible by extracting morbidity data from studies comparing the morbidity of SLNB to that of axillary lymph node dissection. A literature search yielded 13 articles that had data on sensory morbidity at specific time points on pain, numbness or paresthesia from SLNB that used radiotracer and probe or used only dye as a primary method of finding the sentinel node (SN). Of these, 10 utilized LS, while 3 did not utilize LS. By matching the data in studies not employing LS to the studies that did, comparisons regarding the percentage of patients experiencing pain, numbness/paresthesia after SLNB could be reasonably attempted at a cutoff of 9 months. In the 7 studies reporting on pain after 9 months (> 9 months) that used LS (1347 patients), 13.8% of patients reported these symptoms, while in the one study that did not use LS (143 patients), 28.7% of patients reported these symptoms at > 9 months (P < 0.0001). In the 6 studies reporting on numbness and/or paresthesia at > 9 months that used LS (601 patients), 12.5% of patients reported these symptoms, while in the 3 studies that did not use LS (229 patients), 23.1% of patients reported these symptoms at > 9 months (P = 0.0002). Similar trends were also noted for all these symptoms at < or = 9 months. Because of variations in techniques and time of assessing morbidity, direct comparisons between studies are difficult. Nevertheless at a minimum, a clear trend is present: having the LS images and skin markings to assist during SLNB appears to yield more favorable morbidity outcomes for the patients compared to performing SLNB with only the probe or performing SLNB with dye alone. These results are extremely pertinent, as the main reason for performing SLNB itself in the first place is to achieve reduced morbidity.


C-cell follicles of canine thyroid glands studied by PAS reaction and electron microscopy.
Small follicles composed solely of C cells were occasionally observed in large C cell groups of dog thyroid glands. The lumina of C-cell follicles were filled with, or contained peripheral depositions of PAS-positive amorphous material, which was similar in ultrastructural features to thyroglobulin-containing colloid in typical thyroid follicles. This indicates that C cells, in addition to secreting calcitonin, produce a glycoprotein that can be stored in the lumina of the follicles.


Visual acuity and diabetic maculopathy.
We investigated the relationship between visual acuity (VA) and glycosylated hemoglobin, and the presentation and type of diabetic maculopathy retrospectively from 2000 to 2003. Data on 103 eyes of 75 patients were gathered. No statistically significant correlation was found between serum HbA1c with maculopathy type. VA is a poor predictor for the presence or absence of clinically significant diabetic maculopathy and type of maculopathy, emphasizing the importance of systematic screening for and early treatment of maculopathy before visual loss.


Rapid detection of hemoglobin variants by mutagenically separated polymerase chain reaction (MS-PCR).
The detection of molecular defects of hemoglobin variants using mutagenically separated polymerase chain reaction (MS-PCR) was applied in this study. Using different lengths of allele-specific mutagenic primers, normal and mutant alleles of hemoglobin genes were amplified in the same reaction tube. Subsequent gel electrophoresis showed at least one of the two allelic products at the same loci or at least two of the several allelic products at different loci. We employed MS-PCR to test the following hemoglobin variants: Hb Constant Spring (Hb CS), Hb E, Hb G-Taichung, Hb J-Meinung, and Hb Kaohsiung. The results were the same as those obtained by amplified created reaction sites (ACRS) or direct sequencing. We conclude that the MS-PCR provides a rapid and simple alternative to other techniques for mutation detection in hemoglobin variants. Moreover, the principle can be extended to other genetic diseases.


Antitumor agents. Part 232: Synthesis of cyclosulfite podophyllotoxin analogues as novel prototype antitumor agents.
An 11,13-O,O'-cyclosulfite GL-331 analogue (7) was synthesized in six steps from podophyllotoxin and evaluated as a potential antitumor agent. Compound 7 was significantly cytotoxic against human tumor cell lines, but showed no inhibition against human DNA topoisomerase II in vitro. This compound represents a novel prototype of antitumor podophyllotoxin analogues.


The organization of olivo-cerebellar projections in the opossum, Didelphis virginiana, as revealed by the retrograde transport of horseradish peroxidase.
The retrograde transport of horseradish peroxidase was utilized to map olivo-cerebellar projections in the Virginia opossum. The spinal cerebellum (anterior lobe, paramedian lobule and pyramis) receives input from several separate regions in the dorsal accessory nucleus, the medial accessory nucleus and portions of the principal nucleus. Evidence is present for a topographical organization whereby specific regions of the olive project to restricted longitudinal zones. The visual-auditory region of the posterior vermis receives input from small areas within the caudal part of the medial accessory nucleus. From a distinctly separate region of the caudal medial accessory nucleus (as well as the principal nucleus), axons project to the uvula. The vestibulo-cerebellum is the recipient of axons from the cap of Kooy and from two spatially separate regions of the medial accessory nucleus. The cerebellar hemisphere (Crus I and II, lobus simplex) is the target of axons from parts of all three olivary nuclei and it is possible that the projections from the different nuclei are targeted upon separate zones. The paraflocculus was found to receive an input from the rostral part of the medial accessory nucleus and from the principal nucleus. The present results suggest that a distinct olivary region may project to several widely separate areas of the cerebellum, and that one cerebellar region may receive input from several areas of the olive. The organization of the olivocerebellar projection is highly complex, but when considered in light of known inputs to the olive, certain patterns emerge.


Influence of splanchnic nerve on reserpine action in avian adrenal medulla.
Reserpine at three doses (0.05, 0.2, and 0.8 mg/100 g body wt) has been injected intraperitoneally to seven unilaterally splanchnic denervated avian species, in order to ascertain the neural regulation of its action in avian adrenal medulla. The adrenomedullary catecholamines (CA) were estimated at 24 hr after reserpine injection. The findings revealed that reserpine at high doses caused 66-92% depletion of total CA from both the innervated and the denervated glands in all the species investigated. The low dose of reserpine in the passerine (common myna and bulbul) birds resulted in 40-84% reduction of both norepinephrine (NE) and epinephrine (E) from the adrenal glands irrespective of its nerve supply. As a contrast, it depleted 60-80% of both NE and E from the innervated glands as compared to 17-43% reduction from the denervated glands in nonpasserine (woodpecker, parakeet, and koel) birds. Interestingly, in other nonpasserine (pigeon and duck) birds, the low dose of reserpine depleted 66-71% of NE from the innervated glands as compared to only 2-13% reduction from the denervated glands. But in these birds, 60-85% depletion of E, however, occurred independent of neural regulation. The findings suggest that at high doses depletion of CA by reserpine is controlled by some nonneurogenic mechanisms. Generally speaking, at a low dose of reserpine, the splanchnic nerve probably modulates depletion of CA from the adrenal medulla of the nonpasserine birds while the same is governed by some nonneurogenic mechanisms in the passerine birds.


[Effect of colchicine on secretion of alveolar surfactant in intact and regenerating rat lungs].
Electron microscopy showed that colchicin in a total dose of 0.6 mg/100 g body weight of noninbred male white rats inhibited alveolar surfactant secretion into the alveolar lumen in both intact and regenerating lungs. This was associated with a partial or complete alveolar collapse. In the course of inhibition of surfactant secretion, on the apical surface of type II alveolocytes there was an activation of surfactant secretion by way of exocytosis into the interstitial space in the basal part of the cells. Disintegration of the collagenous and elastic fibers as well as disturbance of the membranes occurred in the areas where the material of lipid character and products of its metabolism appeared. A hypothesis is suggested about the possible role of such a disturbance in the character and level of alveolar surfactant secretion in the development of lung abnormality under the effect of the factors inactivating the cytoplasmatic micro tubules.


Effective argon-FRC with a rebreathing technique in children with acute asthma.
In children with asthma the measurement of functional residual capacity (FRC) with standard dilution techniques requires long rebreathing times of the inert gas, i.e., helium, to reach alveolar units with long time constants. A modification of the argon-freon-22 rebreathing technique enables argon to dilute in the readily accessible lung volume and potentially can give a measurement of FRC. However, the rebreathing tests cannot be prolonged for more than 1 min thus allowing argon to reach only the rapidly accessible lung units which are grossly useful in gas exchange (effective FRC, EFRC). The aim of this study was to measure the EFRC and standard lung function in children with acute severe asthma, assess their response to nebulized salbutamol, and evaluate the relations of the EFRC response to baseline spirometric measurements. Twenty-four asthmatic children who were admitted to the hospital with an acute asthma attack had spirometry and triplicate EFRC measurements before and after treatment with nebulized salbutamol. Eighteen patients had repeated the respiratory tests 50 days later when they had fully recovered. A significant proportion (28.9%) of the argon traces obtained at baseline before treatment did not equilibrate. However, in the great majority of these tests the oscillation of the argon traces over the last 3 breaths of the test was < 5% of the simultaneous argon concentration. The EFRC values derived from the argon traces with minimal oscillation (< 5%) were reduced during the acute asthma attack when compared with the paired values obtained after recovery (P = 0.03). The administration of salbutamol caused a fall from the baseline EFRC (P = 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


The surgical management of spinal dural vascular malformations.
Spinal dural arteriovenous fistulas (AVFs) are the most common spinal vascular malformation. The spinal dural AVF is a disease of adults, primarily men, that is a cause of subacute progressive lower extremity myelopathy. The challenge for the optimum management of these patients is in making an accurate, early diagnosis. Definitive surgical intervention by interruption of the intradural draining vein is the treatment of choice.


Flame ionization detection after splitting the water effluent in subcritical water chromatography.
The coupling of subcritical water separation with flame ionization detection (FID) in the split mode has been investigated in this study. In order to keep the FID system stable during subcritical water separation, a Tee union was connected between the separation column and the FID system to split the water flow. The ratio of the water flow to the FID system over the flow-rate to a waste bottle varied depending on the dimension of capillary tubings and the total water flow-rate used. Separations of several carbohydrates, carboxylic acids, and amino acids were performed on commercially available columns using a laboratory-made subcritical water chromatography-FID system. The FID system was very stable in this split mode even at total flow-rate as high as 1.24 ml/min. The linear dynamic range was up to three orders of magnitude and the limit of detection (LOD) ranged from 38 to 111 ng (306-925 ng/microl injected) with split ratios of approximately 1:10 to approximately 1:17 (FID/waste bottle) for several analytes studied. However, the LOD can be significantly lowered by adjusting the dimensions of the restrictors to allow a higher percentage of the total flow to the FID system.


What is the impact of the fluid challenge technique on diagnosis of fluid responsiveness? A systematic review and meta-analysis.
The fluid challenge is considered the gold standard for diagnosis of fluid responsiveness. The objective of this study was to describe the fluid challenge techniques reported in fluid responsiveness studies and to assess the difference in the proportion of 'responders,' (PR) depending on the type of fluid, volume, duration of infusion and timing of assessment. Searches of MEDLINE and Embase were performed for studies using the fluid challenge as a test of cardiac preload with a description of the technique, a reported definition of fluid responsiveness and PR. The primary outcome was the mean PR, depending on volume of fluid, type of fluids, rate of infusion and time of assessment. A total of 85 studies (3601 patients) were included in the analysis. The PR were 54.4% (95% CI 46.9-62.7) where <500 ml was administered, 57.2% (95% CI 52.9-61.0) where 500 ml was administered and 60.5% (95% CI 35.9-79.2) where >500 ml was administered (p = 0.71). The PR was not affected by type of fluid. The PR was similar among patients administered a fluid challenge for <15 minutes (59.2%, 95% CI 54.2-64.1) and for 15-30 minutes (57.7%, 95% CI 52.4-62.4, p = 1). Where the infusion time was ≥30 minutes, there was a lower PR of 49.9% (95% CI 45.6-54, p = 0.04). Response was assessed at the end of fluid challenge, between 1 and 10 minutes, and >10 minutes after the fluid challenge. The proportions of responders were 53.9%, 57.7% and 52.3%, respectively (p = 0.47). The PR decreases with a long infusion time. A standard technique for fluid challenge is desirable.


Predicting ovarian responses to the controlled ovarian hyperstimulation in elderly infertile women using clinical measurements and random forest regression.
During the past decades, the number of elderly infertile women is obviously increasing in China, and more and more of them are likely to seek medical assisted reproductive technologies. As the in vitro fertilization/embryo transfer (IVF/ET) treatment presents special medical and psychological challenges to elderly infertile women, it is extremely helpful to perform the clinical evaluation and outcome prediction regarding IVF/ET outcomes. In this study, we retrospectively collected 12 clinical measurements in prior to the oocyte recovery for 689 elderly infertile patients (≥35 years of old), and used for predicting ovarian responses to the controlled ovarian hyperstimulation based on random forest regression models. Using different predictor sets and 10-fold cross validation approach, the Mean Square Error (±standard deviation) of prediction models varied from 7.56 ± 0.31 to 13.90 ± 0.37 in the training datasets, and the correlation coefficients between observed and predicted values ranged from 0.86 ± 0.02 to 0.72 ± 0.05 in the testing datasets. Among all clinical measurements involved in this study, the preovulatory follicle count (PFC), antral follicle count (AFC), and anti-Müllerian hormone (AMH) were revealed to be the most important features in prediction models. In conclusion, we successfully established the machine learning approach that could help the elderly infertile patients to better understand the most possible outcomes in subjecting to the controlled ovarian hyperstimulation.


Enhanced anticoagulant response to activated protein C in patients with IDDM.
Anticoagulant response to activated protein C (APC) was studied in 40 healthy subjects and 67 patients with insulin-dependent diabetes mellitus (IDDM) using a modified activated thromboplastin time assay. Results are expressed in terms of the APC sensitivity ratio (APC SR). In addition, plasma levels of protein C, total and free protein S (PS), coagulation factors V and VIII, and prothrombin fragment 1+2 (F1+2) were measured. Patients with IDDM and a urinary albumin excretion rate (UAER) < 30 mg/24 h showed a median APC SR of 2.5 (interquartile range 2.3-2.9). In patients with a UAER between 30 and 300 mg/24 h, the median APC SR was 2.7 (2.7-2.9). Both values were significantly greater than the median APC SR of 2.1 (2.0-2.5) observed in healthy control subjects (P < 0.001). Also, the percentage of subjects with an APC SR < or = 2.0 was markedly smaller in both patient groups. Factor V and VIII levels were not significantly different between IDDM patients and healthy subjects. Grouping of IDDM patients according to the APC SR revealed significantly enhanced levels of total PS (P < 0.05) and factor VIII (P < 0.01) in patients with a poor anticoagulant response to APC (APC SR < or = 2.0) compared with those with an APC SR > 2.7. The negative correlation of the APC SR in diabetic patients with both coagulation and anticoagulation factors indicates a complex role of this parameter in regulating the coagulation system in IDDM.


The effect of power frequency high intensity electric fields on implanted cardiac pacemakers.
Thirty-five patients fitted with 16 different pacemaker models (from 6 manufacturers) were exposed to 50 Hz electric fields up to a maximum of 20 kV/m. Four different response patterns were encountered: (1) normal sensing and pacing in all Medtronic and some Vitatron units; (2) reversion to the fixed (interference) rate in all Telectronics, all Pacesetter, some Vitatron and CPI units; (3) slow and irregular pacing in one CPI and in all Cordis units; (4) mixed behavior over a critical range of field strengths in which slow and irregular pacing preceded reversion to fixed-rate, in some Telectronics and Pacesetter units. The field strengths required to induce such behavior varied from unit to unit and from model to model, with Telectronics being the most sensitive. In general, the interference threshold depended on the magnitude and distribution of induced body current relative to the pacemaker as well as field strength and thus varied with patient height, build and posture. While only a small proportion of pacemaker patients are likely to encounter electric fields strong enough to interfere with pacemaker behavior, this possible hazard should be recognized.


Studies on the pathogenesis of fever. XVI. Purification and further chemical characterization of granulocytic pyrogen.
Small quantities of highly purified granulocytic pyrogen have been separated from contaminating proteins by disc electrophoresis in polyacrylamide gel. The biologically active material thus isolated was shown to be electrophoretically homogeneous at pH 9 and pH 3.8. Earlier work on the chemical properties of the pyrogen molecule has been extended to include: (a) estimation of its molecular weight by gel filtration; (b) demonstration of free sulfhydryl groups essential for its biological activity; and (c) evidence that it is not inactivated by exhaustive extraction with ethanolether or n-heptane.


Dissecting demyelination.
The loss of central nervous system myelin and the failure of remyelination by oligodendrocytes contribute to the functional impairment that characterizes diseases such as multiple sclerosis. Why myelin repair fails in multiple sclerosis is currently unclear; however, new understanding of the generation of oligodendrocytes and myelination during development, as well as an increasing understanding of the bases of successful remyelination, are providing new insights and therapeutic targets. We propose that successful myelin repair of the adult CNS recapitulates a sequence of stages that generally correlate with those seen during development, whereas unsuccessful myelin repair results from the perturbation of a critical process in any one of several sequential events. Defining the rate-limiting steps and most vulnerable aspects at each stage of myelin repair will provide logical targets for therapeutic intervention in demyelinating diseases such as multiple sclerosis.


The syndrome of paradoxical worsening of dural-cavernous sinus arteriovenous malformations.
Three patients with dural arteriovenous malformations contiguous with the cavernous sinus had marked worsening of their neuro-ophthalmic symptoms. Such clinical deterioration has been attributed traditionally to increased blood flow through the malformations into the superior ophthalmic vein. However, arteriography in all three patients and magnetic resonance imaging (MR) in two patients demonstrated thrombosis of the entire superior ophthalmic vein. All patients subsequently underwent spontaneous, complete resolution of symptoms. This report describes a clinical syndrome of paradoxical worsening of cavernous sinus malformations and indicates that MR will aid in the management of these vascular malformations.


Bicuculline-induced rhythmic EEG episodes: gender differences and the effects of ethosuximide and baclofen treatment.
There are gender differences in the expression of seizures. We tested rhythmic EEG episodes induced by low doses of bicuculline in rats for gender differences. To verify the validity of these discharges as a model of absence seizures in both male and female rats, we tested the antiabsence drug ethosuximide (ESM) and a gamma-aminobutyric acid (GABA(B))-receptor agonist, baclofen, which may exacerbate absence seizures. Adult rats of both sexes were used. Under general anesthesia, EEG electrodes were implanted over frontal and occipital cortex, and some females were ovariectomized. After recovery, male, intact female rats, and female rats ovariectomized and ovariectomized rats with estradiol replacement were compared for occurrence of rhythmic EEG episodes (approximately 6 cycles/ s) induced by 2.5 mg/kg of bicuculline, s.c. Because of gender differences in sensitivity to bicuculline, further pharmacologic effects of ESM (125 and 250 mg/kg, i.p.) and baclofen (2 mg/kg, i.p.) were tested separately in male (3.0 mg/kg of bicuculline), and female (2.5 mg/kg of bicuculline) rats. After the identical dose of bicuculline, s.c., male and female rats differed in the incidence of rhythmic episodes and in the latency to onset of the first as well as the generalized episode. Female rats with natural or exogenous estrogens (but not ovariectomized rats) developed EEG episodes more often than did males, and this effect could be attributed to the presence of estrogens. ESM pretreatment suppressed the episodes, whereas baclofen enhanced their occurrence, as well as the total duration of episodes without gender-specific differences. The study demonstrates gender differences (related probably to the presence of circulating estrogens) in the susceptibility of rats to develop rhythmic EEG episodes induced by threshold doses of bicuculline. This activity has some features of an acute absence seizure model.


Ginkgolide B protects human pulmonary alveolar epithelial A549 cells from lipopolysaccharide-induced inflammatory responses by reducing TRIM37-mediated NF-κB activation.
The treatment options for acute stroke combined with pulmonary infection are limited. Clinically, there are several therapies to promote blood circulation and dissipate blood stasis; these treatment options include ginkgolide B (GB), which has PAF (platelet activating factor)-inhibiting effects. PAF-receptor (PAF-R) antagonists are used to treat a variety of inflammatory diseases; however, the potential of PAF-R antagonists as a treatment for lung infections remains unclear. The aim of the present study is to investigate the protective effect of GB on lipopolysaccharide-induced inflammatory responses in A549 human pulmonary alveolar epithelial cells (HPAEpiC) in vitro. Cell viability and apoptosis were measured by CCK-8 and flow cytometry. TRIM37, Caspase-3, and NF-κBp65 expression levels were measured by real-time PCR and Western blotting. The release of tumor necrosis factor-α and interleukin-1β was measured by ELISA. The data indicates that GB may reduce TRIM37 expression by antagonizing the PAF-R pathway, thereby inhibiting the activation of nuclear factor-κB and alleviating the inflammatory response of alveolar epithelial cells. This study is the first to provide insight into the therapeutic potential of GB and suggests that clinical application of GB in acute stroke combined with pulmonary inflammation may be efficacious.


Successful treatment of griseofulvin-resistant tinea capitis in infants.
We report on three siblings, children of a farmer, aged 3-8 years, suffering from tinea capitis due to Microsporum canis. Initially, the infection was treated systemically with griseofulvin as well as externally for 5 months without success. Therapy was changed to itraconazole 33 mg per day orally. Laboratory investigations followed every 4-6 weeks. Dependent on the dose per kg body weight the children were cured clinically after 2-5 months and microbiologically after 5-8 months. Thus, itraconazole seems to be superior to conventional treatment with Microsporum canis-induced tinea capitis.


A role for intestinal alkaline phosphatase in the maintenance of local gut immunity.
Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor known to dephosphorylate lipopolysaccharide (LPS); however, the role of IAP in the gut response to luminal bacteria remains poorly defined. We investigated immune responses of wild-type (WT) and IAP-knockout (IAP-KO) mice to LPS and Salmonella typhimurium challenges. Cryostat sectioning and standard indirect immunohistochemical staining for major histocompatibility complex (MHC) class II molecules were performed on liver tissue from WT and IAP-KO mice. WT and IAP-KO mice were orally gavaged with S. typhimurium; bacterial translocation to mesenteric nodes, liver, and spleen was determined by tissue homogenization and plating. In other experiments, WT and IAP-KO mice received intraperitoneal injections of LPS, with subsequent quantification of complete blood counts and serum interleukin (IL)-6 by enzyme-linked immunosorbent assay (ELISA). WT and IAP-KO whole blood were plated and stimulated with LPS and Pam-3-Cys, followed by cytokine assays. Immunohistologic liver examinations showed increased expression of MHC class II molecules in IAP-KO mice. Following S. typhimurium challenge, WT mice appeared moribund compared with IAP-KO mice, with increased bacterial translocation. WT mice had >50% decrease (P<.005) in platelets and 1.8-fold (P<.05) increased serum IL-6 compared with IAP-KO mice in response to LPS injections. IAP-KO whole-blood stimulation with LPS and Pam-3-Cys resulted in increased IL-6 and tumor necrosis factor (TNF)-alpha secretion compared with WT. IAP-KO mice exhibit characteristics consistent with local LPS tolerance. Whole-blood response of IAP-KO mice did not reflect systemic tolerance. These data suggest that IAP is a local immunomodulating factor, perhaps regulating LPS-toll-like receptor 4 (TLR4) interaction between commensal microflora and intestinal epithelium.


Editorial Commentary: Think Twice, Cut Once.
How much of the anterior acetabulum rim needs to be resected to reduce the impingement of the pincer lesion? Hellman et al. present 3 measurements that can be performed on routine radiographs so that the surgeon can preoperatively, intraoperatively, and postoperatively measure the amount resected from the anterior acetabulum rim.


Alexithymia and Cardiac Outcome in Patients at First Acute Coronary Syndrome.
This cohort study was aimed to verify whether subjects at their first acute coronary syndrome (ACS) were more alexithymic than healthy controls (HC) and whether alexithymia can predispose patients with coronary artery disease to new major adverse cardiac events (MACE) during a 24-month follow-up period. The sample included 100 HC with no history of depression or ACS and 304 never depressed patients with a first-ever ACS. A total of 266 patients completed the 2-year follow-up. Patients and HC reported similar Toronto Alexithymia Scale (TAS-20) scores. During the follow-up, 69 (22.69%) patients developed incident depression and 57 (18.75%) developed a new MACE. In a proportional hazard model, developing a first-ever depressive episode, but not alexithymia (hazard ratio = 1.008, 95% confidence interval = 0.984-1.033; p = 0.500), was associated with almost 3 times the risk of a recurrent cardiac event. Incident depression, but not TAS-20 scores, represented risk factor for MACE.


Transcriptomic characterization and construction of M2 macrophage-related prognostic and immunotherapeutic signature in ovarian metastasis of gastric cancer.
Ovarian metastasis (OM) poses a major threat to the outcome of gastric cancer (GC) patients. Recently, immunotherapy emerged as a novel promising therapeutic strategy to treat late-stage GC, whereas its efficacy is influenced by tumor immune microenvironment (TIME). M2 macrophage, a key subset within TIME, plays dual immunosuppressive and pro-tumorigenic roles in cancer progression and is recognized as a potential therapeutic target. However, molecular mechanisms underlying OM remain elusive and the TIME-related prognostic and immunotherapeutic index for these patients is yet to establish. Differential expressed genes (DEGs) between paired normal mucosa, primary GC and OM of patients from Fudan University Shanghai Cancer Center (FUSCC) cohort (n = 6) were identified by transcriptome sequencing, followed by the functional annotation of enriched hallmark pathways of DEGs between them. CIBERSORT was used to profile the relative expression level of 22 immune cell subsets in normal tissues, primary and metastatic tumors, followed by weighted gene coexpression network analysis (WGCNA) uncovering immune cell-correlated gene sets. The intersected genes between DEGs and M2 macrophage-related genes were processed by least absolute shrinkage and selection operator (LASSO) regression analysis to construct a predictive signature, M2GO, which was further validated by training set and test set of The Cancer Genome Atlas-Stomach Adenocarcinoma (TCGA-STAD), GSE62254 and GSE84437 cohorts. GC patients were divided into M2GO-high and -low subgroup according to the optimal cutoff value of the M2GO score. Furthermore, the clinical, molecular and immune features between M2GO-high and -low subgroups were analyzed. Clinical cohorts of immunotherapy were used to validate the predictive value of M2GO in regard to immunotherapy effectiveness. Transcriptomic sequencing and follow-up analyses of triple-matched normal tissues, primary and ovarian metastatic tumors identified distinctive sets of DEGs and enriched immune-, cancer- and metastasis-related pathways between them. Of note, M2 macrophage, a major immunosuppressive and pro-tumorigenic component within TIME, was significantly up-regulated in OMs. WGCNA and LASSO regression were applied to establish a novel OM- and M2 macrophage-related predictive signature, M2GO, based on M2 macrophage-related prognostic genes including GJA1, MAGED1 and SERPINE1. M2GO served as an independent prognostic factor of GC patients. Comprehensive molecular and immune characterization of M2GO-based subgroups uncovered their distinctive features in terms of enriched functional pathways, tumor mutation burden, key immune checkpoints, major regulators of natural immune cGAS-STING pathway, infiltrated subsets of immune cells and tumor immune exclusion/dysfunction (TIDE) score. Notably, the M2GO score was significantly lower in responsive group than non-responsive group (P < 0.05) in clinical cohort of metastatic GC patients undergoing immunotherapy. Transcriptomic characterization of paired normal mucosae, primary and ovarian metastatic tumors revealed their unique molecular and immune features. Follow-up analyses established a novel OM- and M2 macrophage-related signature, M2GO, which served as a promising prognostic and immunotherapeutic biomarker to distinguish the clinical outcome, molecular and immune features of GC patients and predict their differential responses to immunotherapy.


Sitagliptin improves beta-cell function in patients with acute coronary syndromes and newly diagnosed glucose abnormalities--the BEGAMI study.
Newly detected impaired glucose tolerance (IGT) or type 2 diabetes mellitus (T2DM) are common in patients with acute coronary syndrome (ACS; i.e. unstable angina/myocardial infarction) and related to disturbed beta-cell function. The aim of this study is to test the hypothesis that treatment with a dipeptidyl peptidase-4 inhibitor initiated soon after a coronary event improves beta-cell function. Acute coronary syndrome ACS patients with IGT or T2DM (n = 71), screened by oral glucose tolerance test (OGTT) 4-23 days (median 6 days) after hospital admission, were randomly assigned to sitagliptin 100 mg (n = 34) or placebo (n = 37) and treated for a duration of 12 weeks. All patients received lifestyle advice but no glucose-lowering agents other than the study drug. The study end-point was beta-cell function assessed using the insulinogenic index (IGI = ΔInsulin30 /ΔGlucose30 ), derived from an OGTT, and acute insulin response to glucose (AIRg) assessed by a frequently sampled intravenous glucose tolerance test. The IGI and AIRg did not differ at baseline between the sitagliptin and placebo groups (69.9 vs. 66.4 pmol mmol(-1) and 1394 vs. 1106 pmol L(-1) min(-1) respectively). After 12 weeks, the IGI was 85.0 in the sitagliptin and 58.1 pmol/mmol in the placebo group (P = 0.013) and AIRg was 1909 and 1043 pmol L(-1) min(-1) (P < 0.0001) in the sitagliptin and placebo groups respectively. Fasting glucose at baseline was 6.1 mmol L(-1) in sitagliptin-treated patients and 6.0 mmol L(-1) in those who received placebo compared with 5.8 and 5.9 mmol L(-1) respectively, after 12 weeks of treatment. Post load glucose metabolism improved in significantly more sitagliptin-treated patients compared with the placebo group (P = 0.003). Sitagliptin was well tolerated. Sitagliptin improved beta-cell function and glucose perturbations in patients with ACS and newly diagnosed glucose disturbances.


The polymorphic 43Thr bcl-2 protein confers relative resistance to autoimmunity: an analytical evaluation.
We have found a novel polymorphic (Ala43Thr; ACC-->GCC) bcl-2 allele in a Japanese population. An in vitro expression study with a mouse IL-7-dependent pre-B cell line has revealed that inhibition of the programmed cell death function of 43Thr bcl-2 protein is suppressed compared with that of normal 43Ala bcl-2 protein. Since bcl-2 expression in B-lymphoid cells elicits autoimmune disease in mice, we have investigated the possibility of whether a bcl-2 polymorphism has a different susceptibility to autoimmune disease. To evaluate the clinical impact of this polymorphism, the frequency of bcl-2 polymorphism was investigated in 221 children with insulin-dependent diabetes mellitus (IDDM), 237 adults with autoimmune disease (105 with rheumatoid arthritis, 57 with systemic lupus erythematosus, 55 with Sjögren's syndrome, and 20 others), and 290 healthy Japanese children and adults. The frequency of the 43Thr bcl-2 allele, either homozygous or heterozygous, was 14.5% in normal controls, 6.8% (P<0.01) in children with IDDM, and 8.0% (P<0.025) in adults with autoimmune disease. These results suggest that the 43Thr allele of bcl-2 confers resistance to autoimmune disease. The different anti-apoptotic function resulting from the different expression of bcl-2 protein in lymphocytes seems to be associated with the development of autoimmune disease, indicating that the bcl-2 gene affects human autoimmune disease.


Short tip-big difference? First-in-man experience and procedural efficacy of pulmonary vein isolation using the third-generation cryoballoon.
The second-generation cryoballoon (CB2) provides effective and durable pulmonary vein isolation (PVI) associated with encouraging clinical outcome data. The novel third-generation cryoballoon (CB3) incorporates a 40 % shorter distal tip. This design change may translate into an increased rate of PVI real-time signal recording, facilitating an individualized ablation strategy using the time to effect (TTE). Thirty consecutive patients with paroxysmal or short-standing persistent atrial fibrillation underwent CB3-based PVI and were compared to 30 patients treated with the CB2. Individual freeze-cycle duration was set to TTE + 120 s for both groups. A total of 118 (CB3) and 119 (CB2) pulmonary veins (PV) were identified and all PVs successfully isolated utilizing the CB3 and CB2, respectively. The real-time PVI visualization rate was 74 % (CB3) and 40 % (CB2; p = 0.001) and the mean freeze-cycle duration 204 ± 88 s (CB3) and 215 ± 90 s (CB2; p = 0.15). Per individual PV, a shorter mean freeze-duration was found for the CB3 and the right superior PVs (188 ± 92 vs. 211 ± 124 s, p = 0.04) and right inferior PVs (192 ± 75 vs. 200 ± 37 s, p = 0.02). No differences were found for the left-sided PVs. A higher rate of real-time electrical PV recordings is seen using the novel CB3 as compared to CB2, which may facilitate an individualized ablation strategy using the TTE.


A Multicenter Large-Sample Shear Wave Ultrasound Elastographic Study of the Achilles Tendon in Chinese Adults.
The changes in the viscoelasticity of the Achilles tendon are related to tendinopathy. Therefore, constructing a data model in the healthy population is essential to understanding the key factors affecting the viscoelasticity of the Achilles tendon. The purpose of our research was to obtain large sample data, construct a data model, and determine parameters that affect the elastic modulus of the Achilles tendon in healthy Chinese adults. We designed a prospective multicenter clinical trial to evaluate the viscoelasticity of the Achilles tendon by using shear wave elastography. A total of 1165 healthy adult participants from 17 Chinese hospitals were recruited for the assessment. The necessary parameters (age, height, weight, and body mass index) were recorded. The elastic modulus (Young modulus) was obtained from the middle of the Achilles tendon and calculated with feet in naturally relaxed, dorsal, and plantar positions. The thickness and perimeter of the Achilles tendon were measured via cross section on the same site. A multiple linear regression was performed to find the key factors affecting the Young modulus of the Achilles tendon. The Young modulus of the left Achilles tendon in the natural relaxed position followed a normal distribution (P > .05) with a mean ± SD of 374.24 ± 106.12 kPa. The regression equations showed a positive correlation between the Young modulus and weight and a negative correlation between the Young modulus and the circumference or thickness of the left Achilles tendon (P < .05). The Young modulus of the Achilles tendon as measured by shear wave elastography is related to body weight as well as the perimeter or thickness of the tendon.


The histone H3.3K27M mutation in pediatric glioma reprograms H3K27 methylation and gene expression.
Recent studies have identified a Lys 27-to-methionine (K27M) mutation at one allele of H3F3A, one of the two genes encoding histone H3 variant H3.3, in 60% of high-grade pediatric glioma cases. The median survival of this group of patients after diagnosis is ∼1 yr. Here we show that the levels of H3K27 di- and trimethylation (H3K27me2 and H3K27me3) are reduced globally in H3.3K27M patient samples due to the expression of the H3.3K27M mutant allele. Remarkably, we also observed that H3K27me3 and Ezh2 (the catalytic subunit of H3K27 methyltransferase) at chromatin are dramatically increased locally at hundreds of gene loci in H3.3K27M patient cells. Moreover, the gain of H3K27me3 and Ezh2 at gene promoters alters the expression of genes that are associated with various cancer pathways. These results indicate that H3.3K27M mutation reprograms epigenetic landscape and gene expression, which may drive tumorigenesis.