Fluorescent In Situ Hybridization Monitoring and Effect of Detected Early Responses in the Outcome of Patients With Chronic Phase Chronic Myeloid Leukemia: A Report From a Latin American Country.
The cytogenetic hallmark of chronic myeloid leukemia (CML) is the Philadelphia chromosome. Monitoring the response in patients receiving therapy is a standard of care. The present study was conducted to assess the monitoring adherence and reliableness of fluorescent in situ hybridization (FISH) as a monitoring tool and the effect of a complete cytogenetic response (CCyR) assessed by FISH on the prognosis of patients in a chronic phase (CP)-CML cohort. We retrospectively analyzed the data from 63 newly diagnosed CP-CML patients treated with imatinib mesylate at a dose of 400 mg/day as frontline therapy. The clinical data and cytogenetic test results at diagnosis and during monitoring were collected. The cytogenetic monitoring adherence assessment rates were measured. A correlation between chromosome banding analysis (CBA) and FISH was performed. The CCyR assessed by FISH was defined as < 1% BCR-ABL1(+) nuclei. The Kaplan-Meier method was used for overall survival analysis and time-to-event estimates. The cytogenetic monitoring assessment adherence was 50.8% at 3 months, 93.5% at 6 months, 96.7% at 12 months, and 88.6% at 18 months. The Pearson correlation coefficient showed a significantly positive association (r = 0.84; P < .001) between CBA and FISH. The median follow-up duration after imatinib mesylate initiation was 60 months. A CCyR was achieved in 90.4% of patients within the first 18 months of therapy. At 3 months, 31 patients underwent a FISH evaluation, and 13 (41.9%) had achieved a CCyR. The patients who did not achieve a CCyR at 3 months had a significantly inferior probability of 5-year failure-free survival (38% vs. 94%; P = .001) and progression-free survival (80% vs. 100%; P = .043) compared with those with a CCyR. We found improved monitoring adherence compared with the previous reports of Latin American populations. In countries with a high incidence of failure for CBA and a lack of real-time polymerase chain reaction standardization, FISH is a sensitive monitoring tool. In our cohort, patients not achieving an early CCyR, as tested by FISH, were a poor prognosis subgroup with worse rates of failure-free survival and progression-free survival.


Assessing HIV Care Outcomes Among African-Born People Living with HIV in Seattle: An Analysis of the University of Washington Electronic Medical Record.
To examine the relationship between African birth and HIV outcomes and comorbidities among individuals accessing care at the University of Washington. Patients who received a diagnosis of HIV at the University of Washington from 1995 to 2018 were identified. African-born patients were defined as those with recorded birthplace or primary language belonging to an African country. This cohort was compared to all non-African-born patients for initial CD4 count < 200 cells/mL, time from diagnosis to viral suppression, and prevalence of comorbid conditions. We identified 357 African-born and 3710 non-African-born patients. Over the time period, African-born patients were more likely to present with initial CD4 counts < 200 cells/mL (31% vs 19%, p < 0.01), but had shorter time to viral suppression (HR 1.31, [95% CI: 1.14-1.56]). African-born patients had higher rates of hepatitis B and tuberculosis (12% vs. 7% p < 0.01 and 13% vs. 3% p < 0.01). African-born patients living in the Seattle area have better HIV outcomes, but low initial CD4 counts suggest that they are presenting to care late. Increased efforts to engage this population in HIV, hepatitis B, and tuberculosis screening are warranted.


Identification of Multi-kinase Allosteric Inhibitors of Oncogenic Targets EGFR1, PI3K, and BRAF Kinase.
This study aimed to identify promising allosteric inhibitors with the potential to inhibit EGFR1, PI3K, and BRAF kinases as a single agent or in a combination of existing drugs, thus acting as a therapeutic option when traditional drugs fail to give a beneficial response in disease pathology. Upregulation of EGFR1 activates several downstream signaling pathways, resulting in pathophysiological alterations that contribute to cancer. The RAS/RAF/MEK/ERK (MAPK) and PI3K/Akt/mTOR (PI3K/Akt/mTOR) pathways are major downstream signalling partners induced by EGFR1 activation. Despite their vast importance, allosteric FDA-approved drugs targeting EGFR1 and these pathways are not available. The objective of the study is to identify novel multi-kinase small molecules with the potential to inhibit major sites of amplification of cancer signalling pathways, i.e., EGFR1, PI3K/Akt/mTOR, and RAS/RAF/MEK/ERK (MAPK) signalling pathways targeting allosteric sites. In silico methods were used to identify the potential inhibitors using EGFR1, PI3, and BRAF crystal structures complexed with allosteric inhibitors. The potential novel molecules were confirmed for their drug-likeness. Their stability of binding was also confirmed using molecular dynamics simulation studies. To eliminate false negatives, this study used a pharmacophore and structure-based targeting method. The current study was effective in identifying drug-like small molecules, such as ZINC38783966, ZINC01456629, ZINC01456628, and 124173751, 137352549, 137353176, 137352399, 132020316 from ZINC and PubChem database, respectively, with a potential to bind EGFR1 (6DUK), PI3 (4A55) and BRAF (6P3D) at allosteric sites. A 50 ns molecular dynamics investigation also revealed that these potential novel multitarget kinase allosteric inhibitors exhibited stable binding. Alterations in EGFR1, PI3K/Akt/mTOR, and RAS/RAF/MEK/ERK (MAPK) signalling pathways are observed in cancers in high frequency and are also used by viral and environmental toxicants for pathologic purposes. These multi-kinase allosteric inhibitors will provide insight into allosteric drug discovery and deepen our understanding of targeting these pathways, either individually or in combination with orthosteric inhibitors.


Identification of Risk Factors of Recurrence Among Patients With Vulvar Paget Disease Treated With Conservative Surgery.
Vulvar Paget disease (VPD) is a benign disease with high recurrence rates. Standard treatment involves conservative surgery with wide local excision of the lesion. The purpose of the present study is to identify factors that increase the risk of relapse. We conducted a retrospective study and included patients treated with conservative surgery for noninvasive VPD. Cox regression analysis was carried out to assess the independent effect of age, presence of positive margins, tumor size greater than 4 cm, bilateral lesions, and compositive morbidity and pathology on recurrence free survival. Post hoc power analysis was performed with the G-power tool using an α error of 0.05. Overall, 39 patients were included with a median age of 70 years (46-85 years). Of those, 19 patients relapsed within a median duration of 30.5 months (5-132 months). Twelve patients (63%) experienced at least a second relapse. The presence of composite comorbidity significantly affected the interval to recurrence (30.09 vs 71.80 months, p = .032). Univariate Cox regression analysis revealed that the presence of composite pathology features was indicative of a higher risk of recurrence (hazard ratio = -3.71, p = .024). The sample size did not allow for adequate power for this latter finding. Microscopically involved tumor margins and tumor size greater than 4 cm did not predict patients at risk of experiencing relapsing disease. Patients with noninvasive VPD experience high relapse rates. The presence of concurrent benign vulvar pathology may increase these rates, although larger sample sizes are needed to ascertain our findings.


Intraflagellar transport protein 172 is essential for primary cilia formation and plays a vital role in patterning the mammalian brain.
IFT172, also known as Selective Lim-domain Binding protein (SLB), is a component of the intraflagellar transport (IFT) complex. In order to evaluate the biological role of the Ift172 gene, we generated a loss-of-function mutation in the mouse. The resulting Slb mutant embryos die between E12.5 and 13.0, and exhibit severe cranio-facial malformations, failure to close the cranial neural tube, holoprosencephaly, heart edema and extensive hemorrhages. Cilia outgrowth in cells of the neuroepithelium is initiated but the axonemes are severely truncated and do not contain visible microtubules. Morphological and molecular analyses revealed a global brain-patterning defect along the dorsal-ventral (DV) and anterior-posterior (AP) axes. We demonstrate that Ift172 gene function is required for early regulation of Fgf8 at the midbrain-hindbrain boundary and maintenance of the isthmic organizer. In addition, Ift172 is required for proper function of the embryonic node, the early embryonic organizer and for formation of the head organizing center (the anterior mesendoderm, or AME). We propose a model suggesting that forebrain and mid-hindbrain growth and AP patterning depends on the early function of Ift172 at gastrulation. Our data suggest that the formation and function of the node and AME in the mouse embryo relies on an indispensable role of Ift172 in cilia morphogenesis and cilia-mediated signaling.


Beta3-integrin mediates smooth muscle cell accumulation in neointima after carotid ligation in mice.
Pharmacological blockade of beta3-integrins inhibits neointimal lesion formation in nonmouse animal models of arterial injury. In contrast, beta3-integrin-deficient (beta3-/-) mice are not protected from neointimal lesion formation after arterial injury. We investigated this discrepancy in beta3-/- and wild-type (beta3+/+) mice using different models of injury. After disruption of the carotid with a transluminal probe, there was no significant difference in neointimal thickening between beta3-/- and beta3+/+ mice. However, after ligation of the carotid without medial disruption, there was reduced neointimal thickening in beta3-/- mice compared with beta3+/+ mice at intervals up to 3 months. Lesion reduction in beta3-/- mice was associated with fewer intimal smooth muscle cells (SMCs) without a difference in SMC apoptosis or proliferation rate compared with beta3+/+ mice, consistent with reduced SMC migration from the media into the intima of beta3-/- mice. Moreover, combined eccentric medial disruption and ligation of the carotid in beta3-/- mice resulted in neointimal lesion formation only at the site of medial disruption. Transplantation of bone marrow cells harvested from beta3+/+ mice into irradiated beta3-/- mice resulted in reduced neointimal lesion formation after carotid ligation injury, confirming the importance of alpha(v)beta3 and not alpha(IIb)beta3 in the attenuated response. The alpha(v)beta3-integrin mediates intimal SMC accumulation that contributes to neointimal thickening in the setting of arterial ligation.


Biochemical features of dietary chloride deficiency syndrome: a comparative study of 30 cases.
The diagnosis in one infant of a severe state of chloride deficiency after ingesting a diet consisting exclusively of a modified cow milk formula containing only 0.5 mEq/100 kcal chloride ion led to the identification of 30 additional infants (age 2.6 +/- 0.7 months) fed the same commercial formula. The total absence of chloride in the urine was used as a biochemical index of subclinical dietary chloride deficiency. Serum and urine values were studied at diagnosis and 10 to 12 days after chloride replenishment, and compared statistically with the values obtained in a control group of 40 infants (age 2.8 +/- 1.2 months) fed exclusively a different modified formula containing an adequate chloride concentration. The outstanding laboratory abnormalities noted in the infants ingesting a low chloride formula were hypochloremia and metabolic alkalosis. Mean serum concentrations of potassium, urea, creatinine, and uric acid were in the normal range, but group values were statistically different from normal. A previously unreported finding was the demonstration of a significant elevation in the serum concentrations of calcium and phosphate and in the urinary excretions of calcium and magnesium, which persisted after almost complete recovery of the remaining biochemical disturbances. These results indicate the potential risk of nephrocalcinosis after dietary chloride deficiency.


Imaging of brain tissue pH and metabolites. A new approach for the validation of volume-selective NMR spectroscopy.
For the validation of volume-selective 1H and 31P NMR spectroscopy of the brain methods are required that allow high resolution quantitative mapping of tissue pH and metabolites on intact brain slices. The following techniques are proposed for this purpose. Tissue pH is imaged on cryostat sections of in situ frozen brains, using umbelliferone as a fluorescent pH indicator (Csiba et al, Brain Res 289, 334-337 (1983]. Regional tissue ATP content is measured in adjacent cryostat sections, using the luciferine/luciferase system of fireflies for evoking substrate-specific bioluminescence (Kogure and Furones Alonso, Brain Res. 154, 273-284 (1978]. Lactate content is imaged in a similar way by inducing substrate-specific bioluminescence with lactate dehydrogenase and luciferase from vibrio Fischeri (Paschen, J. Cereb. Blood Flow Metab. 5, 609-612 (1985]. The spatial resolution of these techniques is better than 100 mu, as exemplified in experimental brain tumors and brain infarct of cats. The applicability of biochemical mapping for the validation of NMR spectroscopy was tested in a global brain ischemia model of cat by correlating surface coil 31P and 1H spectra with the corresponding regional biochemical data, measured in the sensitive volume of the coil. Correlation coefficients were r = 0.907, 0.852 and 0.924 for pH, lactate and ATP, respectively. These results demonstrate that the biochemical measurements obtained by bioluminescence and fluoroscopic imaging correlate closely with the NMR data and, therefore, are appropriate for the validation of more complex applications, such as volume-selective spectroscopy of brain infarcts or tumors.


Connecting levels of analysis in educational neuroscience: A review of multi-level structure of educational neuroscience with concrete examples.
In its origins educational neuroscience has started as an endeavor to discuss implications of neuroscience studies for education. However, it is now on its way to become a transdisciplinary field, incorporating findings, theoretical frameworks and methodologies from education, and cognitive and brain sciences. Given the differences and diversity in the originating disciplines, it has been a challenge for educational neuroscience to integrate both theoretical and methodological perspectives in education and neuroscience in a coherent way. We present a multi-level framework for educational neuroscience, which argues for integration of multiple levels of analysis, some originating in brain and cognitive sciences, others in education, as a roadmap for the future of educational neuroscience, with concrete examples in mathematical learning and moral education.


Identification of mitochondrial disease genes through integrative analysis of multiple datasets.
Determining the genetic factors in a disease is crucial to elucidating its molecular basis. This task is challenging due to a lack of information on gene function. The integration of large-scale functional genomics data has proven to be an effective strategy to prioritize candidate disease genes. Mitochondrial disorders are a prevalent and heterogeneous class of diseases that are particularly amenable to this approach. Here we explain the application of integrative approaches to the identification of mitochondrial disease genes. We first examine various datasets that can be used to evaluate the involvement of each gene in mitochondrial function. The data integration methodology is then described, accompanied by examples of common implementations. Finally, we discuss how gene networks are constructed using integrative techniques and applied to candidate gene prioritization. Relevant public data resources are indicated. This report highlights the success and potential of data integration as well as its applicability to the search for mitochondrial disease genes.


A new approach to reduce the number of animals used in experimental focal cerebral ischemia models.
We describe a novel experimental set-up that allows biochemical, immunohistochemical and morphometric recording of multiple parameters from a single rat brain. The whole brain was cut (coronal sectioning) in a volumetric manner, and 100 cryo-sections (10 microm) were collected from the region of infarction. By use of a scalpel to dissect the cryosection, crude brain material was obtained from the cortical and basal ganglia areas of ischemic and non-ischemic hemispheres. Material from four 10 microm thick sections of the same animal was pooled. About 30 microg protein lysate was extracted per four sections with various lysis buffers; this sufficed for one biochemical or enzymatic test called "micro-Western-blots" or "micro-zymographies". Scraping brain material from cryosections allows the detection of up to 25 parameters from adjacent brain sections of one single rat brain. Different analysis are possible, we have chosen, e.g. to compare factors affecting the basal lamina of cerebral microvessels like the content of the metalloproteinases-2/-9, their tissue inhibitors, the plasminogen activators, collagen type IV, parameters to test the blood-brain barrier: hemoglobin and the protein of the perfusion solution BSA and the infarction volume. On the basis of these parameters it was possible to compare the interactions of the complex processes in the ischemic brain in the same animal in adjacent sections. Thus, this method increases the validity of data comparisons and reduces significantly the number of animals needed in various experimental settings.


Molecular cloning and genetic characterization of the Saccharomyces cerevisiae NGS1/MRE11 gene.
The Saccharomyces cerevisiae ngs1-1 mutant was previously identified by its enhanced sensitivity to simple DNA-alkylating agents such as methyl methanesulfonate but not to UV. Molecular cloning and sequencing of NGS1 as a putative DNA-alkylation repair gene revealed that it isidentical to MRE11, a gene that is involved in DNA recombinational repair. In order to investigate functional domains of the Mre11 protein, nucleotide-sequence alterations of a number of mre11 mutant alleles, including ngs1-1, mre11-1 (ts), mre11-2, mre11-3 and mre11-58, were determined. Most of these mutations map to the N-terminus ofMre11, emphasizing the importance of this highly conserved domain. The ngs1-1 and mre11-3 mutants carry nonsense mutations resulting in truncated proteins. Missense mutations were found in mre11-1 (ts), mre11-2 and mre11-58, of which mre11-2 and mre11-58 mapped to the conserved phosphoesterase domains, indicating the involvement of these motifs in the formation and/or processing of DNA double-strand breaks. Finally, mitotic-recombination assays show that the mre11 delta mutation enhances inter-chromosomal recombination but decreases the intra-chromosomal deletion frequency. In addition, MRE11 appears to play different roles during spontaneous and alkylation-induced homologous mitotic recombination.


Childhood brain function differences in delinquent and non-delinquent hyperactive boys.
Childhood electrophysiological and clinical measures were obtained in 110 hyperactive (HA) and 76 normal children, who were later followed up as adolescents. Official arrest data were obtained on all subjects and used to measure outcome. The usual interpretation, that the presence of a brain function abnormality suggests a poor prognosis, does not apply to the clinical EEG, EEG spectral and ERP measures obtained on these HA boys. In fact, the converse was found to be true, that is EEG and ERP abnormalities were associated with a good outcome, while normal values of these measures were associated with a poor outcome. Data were presented that suggest that there may be two distinct subgroups of HA boys. The first group was characterized by abnormalities in childhood brain function, abnormal changes in brain function with age, less antisocial and hyperactive behavior in childhood, and absence of delinquency in adolescence. The second group was characterized by normal childhood brain function, normal changes in brain function with age, more antisocial and hyperactive behavior in childhood, and teenage delinquency. Childhood EEG and ERP measures were found to be significantly different in these delinquent and non-delinquent HA groups, while social, familial and cognitive attributes were not. The N2 amplitude of the AERP in delinquent hyperactive (DHA) boys was found to be significantly larger than in the non-delinquent hyperactive (NDHA) boys. This N2 amplitude may prove clinically useful in selecting HA boys for delinquency prevention programs.


Glucose enhances recovery of potassium ion homeostasis and synaptic excitability after anoxia in hippocampal slices.
Hippocampal slices exposed to brief anoxia combined with elevated glucose exhibit greater postanoxic recovery of synaptic transmission. Glucose may have improved recovery of synaptic transmission by enhancing the production of metabolic energy during and after anoxia. This enhancement should provide more ATP for energy-requiring ion transport processes, and lead (1) to a delayed onset of complete depolarization of CA1 pyramidal cells during anoxia (anoxic depolarization) and (2) to greater ion transport activity following anoxia. A delay in anoxic depolarization would protect neurons from damage if the duration of anoxic depolarization was shortened. Greater postanoxic ion transport would allow the re-establishment of ion gradients supportive of neuronal and synaptic excitability. The effects of glucose and anoxia on ion homeostasis and synaptic transmission were examined in rat hippocampal slices exposed to different glucose concentrations (5-20 mM). The duration of anoxic depolarization was held constant so that postanoxic damage related to this duration was controlled. We found that K+ transport and recovery of synaptic transmission after anoxia in hippocampal slices improved as glucose concentration increased. Also, anoxic depolarization was delayed as glucose concentration increased. Thus, added glucose may improve postanoxic recovery of synaptic transmission by better supporting ion transport.


Scalable shape-controlled fabrication of curved microstructures using a femtosecond laser wet-etching process.
Materials with curvilinear surface microstructures are highly desirable for micro-optical and biomedical devices. However, realization of such devices efficiently remains technically challenging. This paper demonstrates a facile and flexible method to fabricate curvilinear microstructures with controllable shapes and dimensions. The method composes of femtosecond laser exposures and chemical etching process with the hydrofluoric acid solutions. By fixed-point and step-in laser irradiations followed by the chemical treatments, concave microstructures with different profiles such as spherical, conical, bell-like and parabola were fabricated on silica glasses. The convex structures were replicated on polymers by the casting replication process. In this work, we used this technique to fabricate high-quality microlens arrays and high-aspect-ratio microwells which can be used in 3D cell culture. This approach offers several advantages such as high-efficient, scalable shape-controllable and easy manipulations.


Afferent connections of the mesencephalic reticular formation: a horseradish peroxidase study in the rat.
The afferent connections of the mesencephalic reticular formation were studied experimentally in the rat by the aid of the retrograde horseradish peroxidase tracer technique. The results suggest that the rostral portion of the mesencephalic reticular formation receives its main input from the cerebral cortex, the zona incerta and the fields of Forel, the central gray substance, the nuclei reticularis pontis oralis and caudalis, and the deep cerebellar nuclei. Substantial input to the same territory of the mesencephalic reticular formation appears to come from the superior colliculus, the substantia nigra, the parabrachial area, the spinal trigeminal nucleus, and the nucleus reticularis gigantocellularis, whereas several other brain structures, among which the locus coeruleus and the raphe complex, seem to represent modest but consistent additional input sources. The afferentation of more caudal portions of the mesencephalic reticular formation appears to conform to the general pattern outlined above with only three exceptions; the cerebral cortex, the deep cerebellar nuclei and the spinal trigeminal nucleus seem to be relatively modest sources of projections to these levels. Considering that the mesencephalic reticular formation is a critical structure in the "ascending activating systems" the present results, confirming and extending those of many other investigators, characterize a set of pathways that seem to be an important part of the anatomical substrate of the sleep-walking cycle.


Low dose rivaroxaban for the management of atherosclerotic cardiovascular disease.
Atherosclerotic cardiovascular disease is characterized by some risk of major adverse events despite the availability of effective medical therapies for secondary prevention. There is emerging evidence suggesting that thrombin partly contributes to this residual risk. In fact, thrombin (i.e., activated coagulation factor II) triggers not only the conversion of fibrinogen to fibrin but also platelet activation and various pathways responsible for pro-atherogenic and/or pro-inflammatory effects through interaction with protease activated receptors. To reduce the risk associated with thrombin activation, oral anticoagulants antagonists of vitamin K showed promise, but were associated with unacceptable bleeding rates. Direct oral anticoagulants targeting the activated factors X and II carry a lower risk of bleeding than vitamin K antagonists. Rivaroxaban, a direct inhibitor of activated factor X approved at the dose of 20 mg once daily for the prevention of thromboembolic events, has been also investigated at a reduced dose of 2.5 mg twice daily in several alternative scenarios of atherosclerotic cardiovascular disease, in combination with standard of care. Current guidelines recommend that low-dose rivaroxaban is given in an adjunct to standard therapy to patients with stable atherosclerosis and acute coronary syndromes at low bleeding risk. Several studies are underway to evaluate its putative benefits in other clinical settings.


Optimizing anti-body drug conjugates and radiopharmaceuticals for precision therapy: The next frontier in precision oncology.
Drug delivery to select therapeutic targets for precision oncology has always been a major challenge in oncology. Anti-body drug conjugates (ADC) and radiopharmaceuticals attempt to overcome the major limitations of chemotherapy or mono-clonal antibody therapy as they provide a highly specific means of attacking the vulnerability of the cancer cell. Currently there are over 100 clinical trials in various stages of development with these agents. Years of improved understanding of ADC technology and addressing previous failures have yielded the current pipeline of ADCs with better design. The near future of ADC developmental therapeutics includes firstly continuing to improve drug design, secondly picking right targets, thirdly balancing efficacy vs toxicity, fourth understanding innate and acquired resistance mechanisms, fifth development of ADCs across pan-cancers as tissue-agnostic therapeutics, and lastly combination strategies. There is considerable excitement in the ADC developmental therapeutics and ADCs including radiopharmaceuticals are here to stay and offer to expand our pipeline of effective drugs to combat cancer.


Post-dispersal predation of Acacia farnesiana seeds by Stator vachelliae (Bruchidae) in Central America.
Post-dispersal seed predation by the bruchid beetle Stator vachelliae was investigated in Santa Rosa National Park, Costa Rica. This insect finds the seeds of the leguminous Acacia farnesiana in the feces of horses, deer, and ctenosaur lizards, the current major dispersers. Patterns of oviposition and pre-adult survival of beetles in the seeds were investigated in a series of experiments using fresh horse dung. S. vachelliae never minded into the dung balls, attacking only those seeds located on the surface. Fresh horse dung did not attract insects more readily than dry dung. The proportion of seeds attacked was not related to their density in a defecation, and was similar in three areas with different densities of the host plant. In a fourth area with no fruiting A. farnesiana shrubs all seeds survived insect predation. Bruchids attacked a greater proportion of seeds at 1 m than at 5 m from the edge of the shrub's crown. Seeds were mainly removed from horse dung by rodents with similar intensity in all areas and at both distances; this seed removal interfered with bruchid oviposition and probably with bruchid survival. S. vachelliae oviposited less frequently on seeds in dung fully exposed to sun. When oviposition on a dung pile was high, the distribution of eggs on the seeds was clumped, suggesting that some seeds were preferred to others. By the end of the dry season, bruchids stopped attacking the seeds. The results show that the fate of both seeds and bruchids is greatly influenced by the location and time of defecation.


[Febrile episodes, headache and limb pain as well as generalized myalgia in a 27-year-old returning male traveller].
Unspecific flu-like symptoms, such as fever, headache and limb pain are encountered very often by general practitioners and in emergency departments. In patients with sepsis and a history of travelling to warmer climates, the differential diagnosis needs to be broader than just commonly encountered viral infections. A 27-year-old Swiss man presented with the symptoms mentioned above after a holiday in the south of France. The pulmonary, hepatic and renal status rapidly deteriorated and the patient required intensive care. The initially suspected diagnosis of leptospirosis could be confirmed serologically during the course of the disease.


The burden of infectious diseases in the Brazilian Southern state of Santa Catarina.
Infectious diseases are still significant causes of deaths in Brazil. The objective of this study was to estimate the burden of selected infectious diseases in the Brazilian Southern state of Santa Catarina in 2011. An ecological study was conducted. The infectious diseases included were HIV/AIDS, tuberculosis, hepatitis B, hepatitis C, Chagas disease, diarrheal diseases and other infectious diseases. Data were collected from official health information systems. Disability Adjusted Life Years (DALY) were estimated by the sum of Years of Life Lost (YLL) and Years Lived with Disability (YLD). 45,237.33 DALYs were estimated, with a rate of 685.46 DALYs per 100,000 population. 92.9% was due to YLL and 7.1% to YLD. Men and the age range of 0-4 years presented higher burden. The highest burden was attributed to HIV/AIDS. There was a high concentration of burden rates in the coast regions of the state. It could be concluded that more than 90% of the burden was attributed to the early mortality component. The highest burden was observed among men, children under 5 years of age and at the coast regions of the state. The highest levels of burden were due to HIV/AIDS.


Common nonmutational NOTCH1 activation in chronic lymphocytic leukemia.
Activating mutations of NOTCH1 (a well-known oncogene in T-cell acute lymphoblastic leukemia) are present in ∼4-13% of chronic lymphocytic leukemia (CLL) cases, where they are associated with disease progression and chemorefractoriness. However, the specific role of NOTCH1 in leukemogenesis remains to be established. Here, we report that the active intracellular portion of NOTCH1 (ICN1) is detectable in ∼50% of peripheral blood CLL cases lacking gene mutations. We identify a "NOTCH1 gene-expression signature" in CLL cells, and show that this signature is significantly enriched in primary CLL cases expressing ICN1, independent of NOTCH1 mutation. NOTCH1 target genes include key regulators of B-cell proliferation, survival, and signal transduction. In particular, we show that NOTCH1 transactivates MYC via binding to B-cell-specific regulatory elements, thus implicating this oncogene in CLL development. These results significantly extend the role of NOTCH1 in CLL pathogenesis, and have direct implications for specific therapeutic targeting.


Effects of high voltage nanosecond electric pulses on eukaryotic cells (in vitro): A systematic review.
For this systematic review, 203 published reports on effects of electroporation using nanosecond high-voltage electric pulses (nsEP) on eukaryotic cells (human, animal, plant) in vitro were analyzed. A field synopsis summarizes current published data in the field with respect to publication year, cell types, exposure configuration, and pulse duration. Published data were analyzed for effects observed in eight main target areas (plasma membrane, intracellular, apoptosis, calcium level and distribution, survival, nucleus, mitochondria, stress) and an additional 107 detailed outcomes. We statistically analyzed effects of nsEP with respect to three pulse duration groups: A: 1-10ns, B: 11-100ns and C: 101-999ns. The analysis confirmed that the plasma membrane is more affected with longer pulses than with short pulses, seen best in uptake of dye molecules after applying single pulses. Additionally, we have reviewed measurements of nsEP and evaluations of the electric fields to which cells were exposed in these reports, and we provide recommendations for assessing nanosecond pulsed electric field effects in electroporation studies.


Help Desk Answers: Do corticosteroids reduce bronchiolitis hospitalizations?
Corticosteroids alone don't decrease hospital admissions or length of stay among children with bronchiolitis. Combining oral dexamethasone and inhaled epinephrine appears to prevent one hospital admission for every 11 patients treated.


Bioassay-guided isolation of cyclooxygenase-2 inhibitory and antioxidant phenylpropanoid derivatives from the roots of Dendropanax dentiger.
The root of Dendropanax dentiger (Harms) Merr. is a traditional Chinese medicine that has been used to treat inflammation-related diseases with little scientific validation. In this study, a bioassay-guided phytochemical investigation of D. dentiger led to the isolation of 19 phenylpropanoid derivatives including one new compound (1) and 18 known ones (2-19). Their structures were elucidated by NMR and HRMS as well as comparison with literature data. The ability of cyclooxygenase-2 (COX-2) inhibition and antioxidant of all isolated compounds were measured in vitro. Chlorogenic acid derivatives (14-19) exhibited outstanding COX-2 inhibitory (IC50 = 5.1-93.4 μM) and antioxidant (IC50 = 13.2-31.9 μM) activities. Moreover, the tight structure-activities relationships were proposed. This is the first report on the COX-2 inhibitory activity of phenylpropanoids and D. dentiger.


Contractile responses of dendrites in the bullfrog retina.
Isolated retinae of the bullfrog were found to "contract" when excited either by electric stimulation of the optic nerve or by light stimulation of the photoreceptors. Involvement of the dendrites of the ganglion and amacrine cells in these contractile responses is suggested.


Research progress of vapor phase corrosion inhibitors in marine environment.
Marine corrosion is a kind of material failure phenomenon caused by the interaction between metal structure and marine environment. As an efficient and simple anti-corrosion method, gaseous corrosion inhibitor can have a good inhibition effect on marine atmospheric corrosion. How to effectively use the compound environment-friendly vapor phase corrosion inhibitor has been widely and efficiently applied in the field of marine atmospheric corrosion, which has become the key to prevent marine atmospheric corrosion. In this paper, we introduce the development history, inhibition mechanism, classification, compounding and structure-activity correlation of vapor phase corrosion inhibitors, and the research prospect of environment-friendly vapor phase corrosion inhibitors in marine environment is prospected.


A new fluorescent imaging of renal inflammation with RCP.
The objective of this study is to design a fluorescent imaging agent with R-Gel, one of the recombinant polymers (RCP), for renal inflammation. The R-Gel based on human type I collagen has multiple Arg-Gly-Asp (RGD) motifs which are ligands for some types of integrin receptors on the cell surface. After intravenous administration of R-Gel labeled by Cy7 of a fluorescent dye to three animal models of nephritis mousse, interstitial nephritis (by using UUO model mice), glomerulonephritis (HIGA mice), and ischemia-reperfusion injured kidney (I/R mice), the extent of fluorescent imaging at the renal inflammation was assessed. The Cy7-labeled R-Gel was accumulated in the inflammation site to a significantly greater extent than in the normal one at 24h after administration. The renal pattern of fluorescent imaging was similar to that of administration anti-Mac1 antibody. Taken together, it is conceivable that the R-Gel was targeted to macrophages infiltrated into the inflammation site of kidney.


Family and species as determinants modulating mineral composition of selected wild-growing mushroom species.
It has been known since the 1970s that differences exist in the profile of element content in wild-growing mushroom species, although knowledge of the role of mushroom species/families as determinants in the accumulation of diverse element remains limited. The aim of this study was to determine the content of 63 mineral elements, divided into six separate groups in the fruit bodies of 17 wild-growing mushroom species. The mushrooms, growing in widely ranging types of soil composition, were collected in Poland in 2018. Lepista nuda and Paralepista gilva contained not only the highest content of essential major (531 and 14,800 mg kg-1, respectively of Ca and P) and trace elements (425 and 66.3 mg kg-1, respectively of Fe and B) but also a high content of trace elements with a detrimental health effect (1.39 and 7.29 mg kg-1, respectively of Tl and Ba). A high content of several elements (Al, B, Ba, Bi, Ca, Er, Fe, Mg, Mo, P, Sc, Ti or V) in L. nuda, Lepista personata, P. gilva and/or Tricholoma equestre fruit bodies belonging to the Tricholomataceae family suggests that such species may be characterised by the most effective accumulation of selected major or trace elements. On the other hand, mushrooms belonging to the Agaricaceae family (Agaricus arvensis, Coprinus comatus and Macrolepiota procera) were characterised by significant differences in the content of all determined elements jointly, which suggests that a higher content of one or several elements is mushroom species-dependent. Graphical abstract.


Contribution to the knowledge of Chinese Meconematinae (Orthoptera: Tettigoniidae) I: One new species of Neocyrtopsis Liu amp; Zhang, 2007 with key to species.
This paper describes one new species of Neocyrtopsis Liu Zhang, 2007 and provides the key to all the species. All type specimens are deposited in Guangxi Normal University.


Finnish experiences of health monitoring: local, regional, and national data sources for policy evaluation.
Finland has a long tradition of gathering information about the health and welfare of the adult population. Surveys and administrative registers form the basis for national and local health monitoring in Finland. Different data sources are used in Finland to develop key indicators, which can be used to evaluate how the national health policy targets have been met in different parts of the country and in different population subgroups. Progress has been shown in chronic disease risk factors, such as smoking reduction. However, some health policy targets have not been met. Socioeconomic health differences, for example, have remained large compared with other European countries. Although data availability for key health indicators is good in Finland, there is a need for wider and more comprehensive use of this information by political decision-makers and healthcare professionals.


Floating spine after pedicle subtraction osteotomy for post-traumatic kyphosis.
Pedicle subtraction osteotomy (PSO) was developed to achieve significant correction of a deformity. It was initially used to correct sagittal plane deformities associated with ankylosing spondylitis, but recently it has also been performed in patients with post-traumatic kyphosis. Our aim was to report a case of a floating spine after PSO for post-traumatic kyphosis. A 50-year-old man was injured after a fall. He had a compression fracture at T12 and an open fracture of the right lower limb. Although he presented with focal back pain, his open fracture was treated first by surgical intervention. The T12 compression fracture was treated conservatively. One year later, he had lower limb numbness and muscle weakness. His imaging demonstrated focal kyphosis on T12 and spinal cord compression. The diagnosis was post-traumatic kyphosis, which was treated with PSO. We performed osteotomy at T12, discectomy and bone graft at T11-T12, and posterior fusion from T10 to L2. One year after PSO, we removed the instruments because he complained of pain around them and found complete bony union between T11 and T12. He immediately experienced worse pain and could not walk or stand for more than 10 min. Imaging showed a floating spine between T12 and L1. He underwent anterior fusion at T12-L1, after which his severe back pain disappeared. This case points out a pitfall of PSO. Although it is a powerful tool for correcting an imbalanced spine, we should recognize its pitfalls and try to avoid them.


Influence of the type of fabric on the collection efficiency of gunshot residues.
Collection efficiency of gunshot residues (GSR) as a function of the type of fabric was evaluated. For this set of experiments, fabrics with differences in their sheddability were exposed to shootings. The samples were then stubbed to collect GSR following a standard procedure for SEM/EDX analysis. The results show a strong relationship between sheddability and collection efficiency, since the collection efficiency is about five-fold higher for leather compared with wool. The main factor explaining this difference is probably the rate of saturation (for instance by fibres) of the carbon tapes used to stub the fabrics. An example originating from a real case supports this hypothesis.


The synthesis of some bridgehead N,O-heterocycles of biological interest.
The preparation of 5-aryl-6-oxa-1-azabicyclo[3,3,0]octanes, 6-aryl-5-oxa-1-azabicyclo[4,3,0]nonanes and 7-aryl-6-oxa-1-azabicyclo[5,3,0]decanes from the corresponding substituted omega-chlorobutyrophenones and aminoalcohols is described; and the ring opening reactions are studied. A preliminary biologic activity study has also been conducted, and demonstrated that the 5-(2,5-dimethyl)phenyl-6-oxa-1-azabicyclo[3,3,0]octane possesses antiinflammatory activity.


A novel ras-related gene family.
We have identified a new family of ras genes, the rho genes, which share several properties with the more classical ras gene family consisting of N-, K-, and H-ras. The rho genes, first isolated from a cDNA library from the abdominal ganglia of Aplysia, encode proteins that share 35% amino acid homology with H-ras. Evolutionarily conserved counterparts of rho have been detected in yeast, in Drosophila, in rat, and in man. Sequence analysis reveals over 85% homology between the human and Aplysia proteins. The ras and rho gene products share several common properties; both are 21,000 daltons, both reveal C-terminal sequences required for membrane attachment, and both show blocks of strong internal homology, suggesting that the two proteins may share common functions but may use these functions in different ways.


Loneliness is strongly linked to depression among older adults, a long term study suggests.
The studyLee SL, Pearce E, Ajnakina O, et al. The association between loneliness and depressive symptoms among adults aged 50 years and older: a 12-year population-based cohort study. Lancet Psychiatry 2021;8:1.To read the full NIHR Alert go to https://evidence.nihr.ac.uk/alert/loneliness-strongly-linked-depression-older-adults/.


Constipation and Postprandial Pain in a Patient With Shortness of Breath.
Incidental image findings may be important for resolving seemingly unrelated symptoms at presentation.


The right heart in heart failure with preserved ejection fraction: insights from cardiac magnetic resonance imaging and invasive haemodynamics.
Recent data indicate that right ventricular systolic dysfunction (RVSD) by cardiac magnetic resonance imaging (CMR) is a strong predictor of outcome in heart failure. However, the prognostic significance of RVSD by CMR in heart failure with preserved ejection fraction (HFpEF) is unknown. We prospectively enrolled 171 HFpEF patients who underwent CMR in addition to invasive and non-invasive testing. RVSD, defined as right ventricular (RV) EF <45% by CMR, was present in 33 (19.3 %) patients. Patients were followed for 573 ± 387 days, during which 41 had a cardiac event. Patients with RVSD presented with more frequent history of AF (P = 0.038), significantly higher resting heart rate (P = 0.009), shorter 6-min walk distance (P = 0.036), and higher NT-pro BNP serum levels (P < 0.001), and were more symptomatic (P < 0.001). With respect to haemodynamic parameters, RVSD was associated with respect to haemodynamic parameters, RVSD was associated with higher diastolic pulmonary artery pressure (P = 0.045), with higher pulmonary vascular resistance (P = 0.048), higher transpulmonary gradient (P = 0.042), and higher diastolic pulmonary vascular pressure gradient (P = 0.007). In the multivariable Cox analysis, RVSD (P < 0.001) remained significantly associated with cardiac events, in addition to diabetes (P = 0.011), 6-min walk distance (P = 0.018), and systolic pulmonary artery pressure (P = 0.003). Although HFpEF is considered a disease of the left ventricle, respective imaging parameters are not related to outcome. In contrast, RVSD by CMR is independently associated with mortality and clinical status in these patients, and provides a useful tool for risk stratification.


Activity, Activity Personalization, and Well-Being in Nursing Home Residents With and Without Cognitive Impairment: An Integrative Review.
Engagement in personally relevant and pleasant activity is a facet of many theories of well-being. This integrated review and narrative synthesis explored the hypothesis that activity participation improves well-being for nursing home residents. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, we searched MEDLINE, PsycINFO, Academic Search Complete, Psychology and Behavioral Sciences Collection, Embase, and CINAHL for research published between 2006 and 2018. We included peer-reviewed, English-language studies of nursing-home residents, with interventions focused on activities and on well-being or affect outcomes. After screening, the search yielded 45 studies: 15 reviews of specific activities and 30 empirical articles. We found consistent support for tailored activity interventions and less consistent support for specific or generic activity interventions with the possible exception of music therapy. Research focused on specific activity types had methodological limitations and confounds with activity preferences. Participation in activities may improve well-being in residents of nursing homes. Tailored activities are likely to be superior to those provided indiscriminately to all residents. Improving quality of life in long-term care should include opportunities to engage in activities; those most effective will be tailored to individuals and no single activity will be effective for everyone.


Peripheral antinociception and anti-edematogenic effect of a sulfated polysaccharide from Acanthophora muscoides.
Sulfated polysaccharides from red marine algae have presented a variety of potentially therapeutic biological effects, however, their antinocicpetive and anti-inflammatory properties are not well understood. Male Swiss mice were pretreated with a sulfated polysaccharidic fraction obtained from the marine alga Acanthophora muscoides (AmII) (1, 3 or 9 mg/kg, iv) 30 min prior to either receiving an injection of 0.8% acetic acid or 1% formalin or prior to a thermal stimulus. AmII (1, 3 or 9 mg/kg, sc) was evaluated on carrageenan-, dextran- bradykinin-, histamine- and serotonin-induced rat paw edema models. AmII (500 μg, sc) was also injected into the paw. Additionally, mice were treated with the total sulfated polysaccharides from A. muscoides (Am-TSP) (20 mg/kg, ip) for 14 days. AmII reduced the number of acetic acid-induced writhes and licking time in the second phase of the formalin test, but it did not alter the response latency in the hot plate test, suggesting that its antinociceptive action occurs through a peripheral mechanism. AmII did not reduce carrageenan-induced paw edema and MPO activity. However, it reduced dextran-, histamine- and serotonin-induced paw edemas, but not bradykinin-induced edema, suggesting that histamine is the major target of AmII anti-edematogenic activity. AmII injected into the paw did not evoke local edema. Furthermore, Am-TSP induced no consistent signs of systemic damage, as revealed by body mass, organs wet weight and by biochemical, hematological and histopathological analyses. AmII has important antinociceptive and anti-inflammatory properties and represents an important therapeutic agent warranting future studies.


Release of cell surface-associated basic fibroblast growth factor by glycosylphosphatidylinositol-specific phospholipase C.
Heparan sulfate proteoglycans (HSPG) are ubiquitous constituents of mammalian cell surfaces and most extracellular matrices. A portion of the cell surface HSPG is anchored via a covalently linked glycosyl-phosphatidylinositol (Pl) residue, which can be released by treatment with a glycosyl-Pl specific phospholipase C (Pl-PLC). We report that exposure of bovine aortic endothelial and smooth muscle cells to Pl-PLC resulted in release of cell surface-associated, growth-promoting activity that was neutralized by antibasic fibroblast growth factor (bFGF) antibodies. Active bFGF was also released by treating the cells with bacterial heparitinase. Under the same conditions there was no release of mitogenic activity from cells (BHK-21, NIH/3T3, PF-HR9) that expressed little or no bFGF, as opposed to Pl-PLC-mediated release of active bFGF from the same cells transfected with the bFGF gene. The released bFGF competed with recombinant bFGF in a radioreceptor assay. Addition of Pl-PLC to sparsely seeded vascular endothelial cells resulted in a marked stimulation of cell proliferation, but there was no mitogenic effect of Pl-PLC on 3T3 fibroblasts. Studies with exogenously added 125I-bFGF revealed that about 6.5% and 20% of the cell surface-bound bFGF were released by treatment with Pl-PLC and heparitinase, respectively. Both enzymes also released sulfate-labeled heparan sulfate from metabolically labeled 3T3 fibroblasts. Pl-PLC failed to release 125I-bFGF from the subendothelial extracellular matrix (ECM), as compared to release of 60% of the ECM-bound bFGF by heparitinase. Our results indicate that 3-8% of the total cellular content of bFGF is associated with glycosyl-Pl anchored cell surface HSPG. This FGF may exert both autocrine and paracrine effects, provided that it is released by Pl-PLC and adequately presented to high affinity bFGF cell surface receptor sites.


Myopericarditis Complicated With Cardiac Tamponade Due to Campylobacter fetus Infection.
About 90% of cases of acute pericarditis have an idiopathic or viral etiology. In some cases, it is possible to identify high-risk patients for whom hospital admission and specific etiology research are mandatory for adequate treatment. Bacterial pericarditis is uncommon and responsible for less than 1% of cases. Only 10 cases of pericarditis due to Campylobacter fetus have been documented worldwide. This case highlights the importance of good cardiac imaging, with the right clinical and microbiology-integrated approach in high-risk cases of pericardial disease.


Enrichment of epidermal stem cells of rats by Vario magnetic activated cell sorting system.
Epidermal stem cells (ESCs) play an important role in skin homeostasis, wound repair, and tumorigensis which have great potential in scientific research and clinical application. So, the efficient isolation of these infrequent stem cells is very important for researchers to solve the problem of low purity and insufficient quantity of stem cells in vitro. The aim of this study was to investigate a method for the enrichment of ESCs by magnetic activated cell sorting system. The isolation strategy was CD71 depletion followed by α6-integrin positive selection. The percentage of α6(bri)CD71(dim) cells in isolated cells was 94.59%. Transmission electron microscopy results revealed that α6(bri) CD71(dim) cells exhibited some typical characteristics like progenitor cells, such as big nucleus, obvious nucleolus, large nuclear-cytoplasm ratio, and few organelles in cytoplasm. When cultured in vitro, the α6(bri)CD71(dim) cells had greater proliferating potential and higher colony-forming ability, and high levels of epidermal stem cell markers were expressed in our positive cells. ESCs have been successfully isolated from neonatal epidermis using Vario MACS and cultured in vitro. This isolation method is simple, fast, and inexpensive, providing an important tool for tissue engineering and cell transplantation studies.


Novel species of Cercospora and Pseudocercospora (Capnodiales, Mycosphaerellaceae) from Australia.
Novel species of Cercospora and Pseudocercospora are described from Australian native plant species. These taxa are Cercospora ischaemi sp. nov. on Ischaemum australe (Poaceae); Pseudocercospora airliensis sp. nov. on Polyalthia nitidissima (Annonaceae); Pseudocercospora proiphydis sp. nov. on Proiphys amboinensis (Amaryllidaceae); and Pseudocercospora jagerae sp. nov. on Jagera pseudorhus var. pseudorhus (Sapindaceae). These species were characterised by morphology and an analysis of partial nucleotide sequence data for the three gene loci, ITS, LSU and EF-1α. Recent divergence of closely related Australian species of Pseudocercospora on native plants is proposed.


Neuronal distribution of spatial in the developing cerebellum and hippocampus and its somatodendritic association with the kinesin motor KIF17.
We identified the Spatial (Stromal Protein Associated with Thymii and Lymph-node) gene from an adult thymus mouse library of cDNA clones. By RT-PCR, we reported that Spatial was highly expressed in restricted areas of the central nervous system. Here, we characterize the precise cellular localization of Spatial during mouse brain development in the cerebellum, hippocampus and cortex. Five different transcript isoforms have been described for Spatial and among those, only Spatial-epsilon and -beta present a tightly controlled expression. In the cerebellum, Spatial expression is detected in the external precursor granular layer and persists as these cells migrate and differentiate to form the internal granular layer. It is also expressed in differentiating Purkinje cells with a specific somatodendritic distribution. Spatial expression in the hippocampus is spatially and temporally regulated: it is first expressed in the CA3 field, then in CA1 and later in the dentate gyrus. Interestingly, Spatial-beta expression tightly overlaps with the beginning of neuronal differentiation in both structures. Using cultured hippocampal neurons, we show that Spatial also exhibits a somatodendritic distribution and it is concentrated in some synaptic regions. Moreover, the vesicle-like cellular distribution of Spatial protein in dendrites is similar to that described for the kinesin motor protein KIF17. Immunofluorescence analyses show that Spatial colocalizes with KIF17 in dendrites of hippocampal neurons in primary culture. Additionally, coimmunoprecipitation experiments of endogenous proteins from hippocampus confirmed that Spatial and KIF17 physically interact. These findings suggest that Spatial may play a role in neuronal morphogenesis and synaptic plasticity through its interaction with the kinesin motor KIF17 in dendrites.


Modulation of unitary glutamatergic synapses by neurotrophin-4/5 or brain-derived neurotrophic factor in hippocampal microcultures: presynaptic enhancement depends on pre-established paired-pulse facilitation.
The neurotrophins, nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin-4/5, have--in addition to their known effects as neuronal survival factors--recently been found to modulate synaptic transmission in the rat hippocampus and neocortex. Using standard whole-cell patch-clamp recordings, we have now investigated the acute effects of brain-derived neurotrophic factor and neurotrophin-4/5 on unitary (i.e. single cell activated) glutamatergic synaptic connections in microcultures of postnatal rat hippocampal neurons. We show that, in approximately 30% of the cells, glutamatergic synaptic transmission is enhanced to 170 +/- 52% (neurotrophin-4/5, 100 ng/ml) and 143 +/- 35% (brain-derived neurotrophic factor, 100 ng/ml) of control values, respectively. The enhancement is abolished in the presence of the specific Trk tyrosine kinase inhibitor k252a (200 nM). Depending on the particular cell investigated, the enhancement consisted of transient and sustained components in varying quantities. A minority of neurons (10%) showed a depression of glutamatergic synaptic transmission to 64 +/- 14% (brain-derived neurotrophic factor) and 61 +/- 11% of control (neurotrophin-4/5). The enhancement of unitary glutamatergic synaptic transmission is mediated predominantly by presynaptic modifications, as is evident from (i) the concomitant decrease in paired-pulse facilitation, (ii) the concomitant increase in the variance of the evoked unitary synaptic currents and (iii) the enhanced miniature excitatory postsynaptic/autaptic current frequencies that could be observed in the absence of an effect on miniature excitatory postsynaptic/autaptic current amplitudes. Finally, we show that the successful enhancement of synaptic transmission by neurotrophin-4/5 critically depends on the degree of paired-pulse facilitation prior to the start of neurotrophin application, with autapses/synapses initially showing a higher degree of paired-pulse facilitation being enhanced more effectively. Taken together, these results suggest that the brain-derived neurotrophic factor- and neurotrophin-4/5-mediated enhancement of unitary glutamatergic synaptic transmission in hippocampal cultures results predominantly from a presynaptic modulation of transmitter release, and this modulation could participate in the neurotrophin-dependent modification of glutamatergic synaptic transmission in the hippocampus in situ.


A catalyst-free co-reaction system of long-lasting and intensive chemiluminescence applied to the detection of alkaline phosphatase.
A catalyst-free co-reaction luminol-H2O2-K2S2O8 chemiluminescence (CL) system was developed, with long-life and high-intensity emission, and CL emission lasting for 6 h. A possible mechanism of persistent and intense emission in this CL system was discussed in the context of CL spectra, cyclic voltammetry, electron spin resonance (ESR), and the effects of radical scavengers on luminol-H2O2-K2S2O8 system. H2O2 and K2S2O8 co-reactants can promote each other to continuously generate corresponding radicals (OH•, 1O2, O2•-, SO4•-) that trigger the CL emission of luminol. H2O2 can also be constantly produced by the reaction of K2S2O8 and H2O to further extend the persistence of this CL system. CL emission can be quenched via ascorbic acid (AA), which can be generated through hydrolysis reaction of L-ascorbic acid 2-phosphate trisodium salt (AAP) and alkaline phosphatase (ALP). Next, a CL-based method was established for the detection of ALP with good linearity from 0.08 to 5 U·L-1 and a limit of detection of 0.049 U·L-1. The proposed method was used to detect ALP in human serum samples.


Giant prostate carcinoma: computed tomography findings and review of previous reports.
Two patients with massive prostate adenocarcinoma were evaluated by several imaging modalities. Computed tomography provided excellent anatomic delineation of the tumors. In one patient, computed tomography demonstrated significant tumor regression following orchiectomy. Giant prostate carcinoma should be considered in the differential diagnosis of a large, inhomogeneous pelvic mass.


Low-frequency characteristics of intracellularly recorded receptor potentials in guinea-pig cochlear hair cells.
Intracellular receptor potentials were recorded from inner and outer hair cells in response to low-frequency tones, from the basal, high-frequency region of the guinea-pig cochlea. The receptor potentials recorded from inner hair cells are asymmetrical about the resting membrane potential with the depolarizing phase, which corresponds to rarefaction in sound pressure, exceeding the phase of hyperpolarization by a factor of about 3. It was found that the relationship between the peak-to-peak voltage responses and sound pressure level could be described by rectangular hyperbolae. When the frequency of the sound stimulus was progressively increased from 100 Hz to 4 kHz, the 'periodic' (a.c.) component of the receptor potential was attenuated with respect to the 'continuous' (d.c.) component. The characteristics of the inner hair cells could be described by two stages of low-pass filtering, with one of the filters having the same corner frequency as the electrical time constants which varied in different cells between 178 and 840 Hz. Receptor potentials recorded intracellularly from two morphologically identified outer hair cells were symmetrical about the resting membrane potential (about -65-70 mV) and had a maximal amplitude of only 5 mV at frequencies and intensities which yield 20-30 mV voltage responses from inner hair cells. No d.c. component receptor potentials were recorded in response to high-frequency tones. Phase and amplitude measurements were made from receptor potentials from inner hair cells, and from 'cochlear microphonic potentials' which were recorded from the organ of Corti and scala tympani. The phase of depolarization in both potentials was associated with displacement of the basilar membrane towards the scala vestibuli. The phase of the intracellular receptor potentials leads the cochlear microphonic by about 90 degrees and the sound pressure by about 180 degrees at frequencies below 100 Hz. Above this frequency the phase lead progressively declines and at higher frequencies becomes a phase lag. These phase relationships indicate that inner hair cells respond to the velocity of the basilar membrane at frequencies below 200-600 Hz, and to its displacement above this, and that the voltage responses of the inner hair cells are limited by their membrane time constants. It is suggested that outer hair cells respond to basilar membrane displacement throughout their frequency range. It is shown that, with respect to frequency, the different growth rates of the cochlear microphonic potentials and inner hair cell receptor potentials, and the dominance of cochlear microphonic potentials in the organ of Corti, result in an effective electrical interaction between inner hair cells and cochlear microphonic potentials.


The influence of age on the natural history of unipolar depression when treated with electroconvulsive therapy.
The influence of age on the natural history of unipolar depression when treated with electroconvulsive therapy (ECT) was studied using a naturalistic/archival study design. A sample of 125 patients who received no somatic treatment were compared with 128 patients who all received a course of ECT with at least four treatments. Patients were separated according to age at admission. Treated patients, aged 40 or older, who were clearly remitters showed no differences in previous episodes, subsequent episodes, subsequent hospitalizations, or likelihood of experiencing a period of full recovery when compared with a similar group of untreated patients. Hospitalization greater than 1 year and chronicity were significantly more common in the untreated older subjects. Treated patients aged 39 or younger, who also were clearly remitters, showed significant increases in subsequent episodes and subsequent hospitalizations when compared with a group of depressed patients of similar age who received no somatic treatment. Hospitalization greater than 1 year was also more common in the untreated younger patients. ECT clearly reduces the rate of chronicity in older patients but may be associated with an increase in episodes after treatment in the younger population.


Polydopamine-coated thalidomide nanocrystals promote DSS-induced murine colitis recovery through Macrophage M2 polarization together with the synergistic anti-inflammatory and anti-angiogenic effects.
High levels of proinflammatory cytokines, macrophage polarization status and immune-mediated angiogenesis play pivotal roles in the pathogenesis of inflammatory bowel disease (IBD). Thalidomide, an anti-inflammatory, immunomodulatory and antiangiogenic agent, is used off-label for treatment of IBD. The therapeutic potential of thalidomide is limited by its poor solubility and side effects associated with its systemic exposure. To address these issues and promote its therapeutic effects on IBD, thalidomide nanocrystals (Thali NCs) were prepared and coated with polydopamine (PDA), a potential macrophage polarization modulator, to form PDA coated Thali NCs (Thali@PDA). Thali@PDA possessed a high drug loading and displayed average particle size of 764.7 ± 50.30 nm. It showed a better anti-colitis effect than bare thalidomide nanocrystals at the same dose of thalidomide. Synergistic effects of polydopamine on anti-inflammatory and anti-angiogenic activities of thalidomide were observed. Furthermore, PDA coating could direct polarization of macrophages towards M2 phenotype, which boosted therapeutic effects of Thali@PDA on IBD. Upon repeated dosing of Thali@PDA for one week, symptoms of IBD in mice were significantly relieved, and histomorphology of the colitis colons were normalized. Key proinflammatory cytokine levels in the inflamed intestines were significantly decreased. Toxicity study also revealed that Thali@PDA is a safe formulation.


Risk of developing another basal cell carcinoma. A 5-year prospective study.
This 5-year prospective study of 1000 people, who had one basal cell carcinoma treated, found that 36% develop a second basal cell carcinoma. The most important risk factors are a skin complexion with inability to tan and frequent sun exposure. There was no significant correlation between multiple basal cell carcinomas and sex or age.


Quantitative and time-course analysis of microbial degradation of 1H,1H,2H,2H,8H,8H-perfluorododecanol in activated sludge.
A methylene group in the fluorinated carbon backbone of 1H,1H,2H,2H,8H,8H-perfluorododecanol (degradable telomer fluoroalcohol, DTFA) renders the molecule cleavable by microbial degradation into two fluorinated carboxylic acids. Several biodegradation products of DTFA are known, but their rates of conversion and fates in the environment have not been determined. We used liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) to quantitatively investigate DTFA biodegradation by the microbial community in activated sludge in polyethylene terephthalate (PET) flasks, which we also determined here showed least adsorption of DTFA. A reduction in DTFA concentration in the medium was accompanied by rapid increases in the concentrations of 2H,2H,8H,8H-perfluorododecanoic acid (2H,2H,8H,8H-PFDoA), 2H,8H,8H-2-perfluorododecenoic acid (2H,8H,8H-2-PFUDoA), and 2H,2H,8H-7-perfluorododecenoic acid and 2H,2H,8H-8-perfluorododecenoic acid (2H,2H,8H-7-PFUDoA/2H,2H,8H-8-PFUDoA), which were in turn followed by an increase in 6H,6H-perfluorodecanoic acid (6H,6H-PFDeA) concentration, and decreases in 2H,2H,8H,8H-PFDoA, 2H,8H,8H-2-PFUDoA, and 2H,2H,8H-7-PFUDoA/2H,2H,8H-8-PFUDoA concentrations. Accumulation of perfluorobutanoic acid (PFBA), a presumed end product of DTFA degradation, was also detected. Our quantitative and time-course study of the concentrations of these compounds reveals main routes of DTFA biodegradation, and the presence of new biodegradation pathways.


ZnO, TiO(2), SiO(2,) and Al(2)O(3) nanoparticles-induced toxic effects on human fetal lung fibroblasts.
This study aims to investigate and compare the toxic effects of four types of metal oxide (ZnO, TiO(2), SiO(2,) and Al(2)O(3)) nanoparticles with similar primary size (∼20 nm) on human fetal lung fibroblasts (HFL1) in vitro. The HFL1 cells were exposed to the nanoparticles, and toxic effects were analyzed by using MTT assay, cellular morphology observation and Hoechst 33 258 staining. The results show that the four types of metal oxide nanoparticles lead to cellular mitochondrial dysfunction, morphological modifications and apoptosis at the concentration range of 0.25-1.50 mg/mL and the toxic effects are obviously displayed in dose-dependent manner. ZnO is the most toxic nanomaterials followed by TiO(2), SiO(2), and Al(2)O(3) nanoparticles in a descending order. The results highlight the differential cytotoxicity associated with exposure to ZnO, TiO(2), SiO(2), and Al(2)O(3) nanoparticles, and suggest an extreme attention to safety utilization of these nanomaterials.


Assessing Public's Attitudes Towards Euthanasia and Assisted Suicide of Persons With Dementia Based on Their Advance Request: An Experimental Survey of US Public.
Advance request euthanasia and/or assisted suicide (AR-EAS) in persons with dementia is highly controversial. Results of typical public opinion surveys may not reflect the ethical and practical issues involved in the practice. We tested the impact of incorporating such issues in the assessment of public attitudes toward legalization of AR-EAS. Online survey (April 27-30, 2020) of 1,711 adults recruited via CloudResearch PrimePanel, matched to U.S. population in age, sex, race and/or ethnicity, education, household income, and political affiliation. After assessing initial attitudes toward legalization of AR-EAS, respondents viewed one of six randomly assigned scenarios depicting an ethical or practical issue in AR-EAS; acceptability of EAS in each scenario as well as general attitudes toward AR-EAS legalization were then elicited. Approximately 54.4% initially agreed/strongly agreed with AR-EAS legalization; agreement was associated with lower dementia quality-of-life rating, younger age, not being religious, liberal politics, and $75,000-$99,999 income range. After viewing the scenarios, a minority in each scenario arm found the AR-EAS depicted acceptable (20.7%-39.1%; p<0.0001 for all six arms, in comparison with initial legalization question response). Support (agree/strongly agree) for AR-EAS legalization after reading specific scenarios was generally lower (range 36.5%-49.3%; p≤0.0002); change in support for legalization was associated with initial support for legalization, acceptability of AR-EAS in the scenarios, dementia quality-of-life ratings, and race. Informing the public of the ethical and practical complexities in AR-EAS may have significant effects on their attitudes toward legalization. Future surveys should ensure that the public's views reflect sufficient exposure to these complexities.


Correlating the above- and belowground genotype of Pinus pinaster trees and rhizosphere bacterial communities under drought conditions.
Increasing temperatures along with severe droughts are factors that may jeopardize the survival of the forests in the Mediterranean basin. In this region, Pinus pinaster is a common conifer species, that has been used as a model species in evolutionary studies due to its adaptive response to changing environments. Although its drought tolerance mechanisms are already known, knowledge about the dynamics of its root microbiota is still scarce. We aimed to decipher the structural (bacterial abundance), compositional, functional and associative changes of the P. pinaster rhizosphere bacterial communities in spring and summer, at DNA and RNA level (environmental DNA, live and dead cells, and those synthesizing proteins). A fundamental aspect of root microbiome-based approaches is to guarantee the correct origin of the samples. Thus, we assessed the genotype of host needles and roots from which rhizosphere samples were obtained. For more than 50% of the selected trees, genotype discrepancies were found and in three cases the plant species could not be determined. Rhizosphere bacterial communities were homogeneous with respect to diversity and structural levels regardless of the host genotype in both seasons. Nonetheless, significant changes were seen in the taxonomic profiles depending on the season. Seasonal changes were also evident in the bacterial co-occurrence patterns, both in DNA and RNA libraries. While spring communities switched to more complex networks, summer populations resulted in more compartmentalized networks, suggesting that these communities were facing a disturbance. These results may mirror the future status of bacterial communities in a context of climate change. A keystone hub was ascribed to the genus Phenylobacterium in the functional network calculated for summer. Overall, it is important to validate the origin and identity of plant samples in any plant-microbiota study so that more reliable ecological analyses are performed.


[Acral necrosis as a complication of urosepsis].
Sepsis is the third most common cause of death in Germany. Every fourth patient with sepsis has urosepsis. Even if substantial therapeutic progress has been made, sepsis remains a severe condition with high morbidity and mortality that requires rapid interdisciplinary measures. Besides life-threatening complications, acral necrosis as presented here can occur as a result of disseminated intravascular coagulation and severe microcirculatory disorders.


Relationships of online exhaled, offline exhaled, and ambient nitric oxide in an epidemiologic survey of schoolchildren.
Field measurements of exhaled nitric oxide (FeNO) and ambient nitric oxide (NO) are useful to assess both respiratory health and short-term air pollution exposure. Online real-time measurement maximizes data quality and comparability with clinical studies, but offline delayed measurement may be more practical for large epidemiological studies. To facilitate cross-comparison in larger studies, we measured FeNO and concurrent ambient NO both online and offline in 362 children at 14 schools in 8 Southern California communities. Offline breath samples were collected in bags at 100 ml/s expiratory flow with deadspace discard; online FeNO was measured at 50 ml/s. Scrubbing of ambient NO from inhaled air appeared to be nearly 100% effective online, but 50-75% effective offline. Offline samples were stored at 2-8 degrees C and analyzed 2-26 h later at a central laboratory. Offline and online FeNO showed a nearly (but not completely) linear relationship (R(2)=0.90); unadjusted means (ranges) were 10 (4-94) and 15 (3-181) p.p.b., respectively. Ambient NO concentration range was 0-212 p.p.b. Offline FeNO was positively related to ambient NO (r=0.30, P<0.0001), unlike online FeNO (r=0.09, P=0.08), indicating that ambient NO artifactually influenced offline measurements. Offline FeNO differed between schools (P<0.001); online FeNO did not (P=0.26), suggesting artifacts related to offline bag storage and transport. Artifact effects were small in comparison with between-subject variance of FeNO. An empirical statistical model predicting individual online FeNO from offline FeNO, ambient NO, and lag time before offline analysis gave R(2)=0.94. Analyses of school or age differences yielded similar results from measured or model-predicted online FeNO. Either online or offline measurement of exhaled NO and concurrent ambient NO can be useful in field epidemiology. Influence of ambient NO on exhaled NO should be examined carefully, particularly for offline measurements.


Population pharmacokinetics and dosing implications for cobimetinib in patients with solid tumors.
To characterize cobimetinib pharmacokinetics and evaluate impact of clinically relevant covariates on cobimetinib pharmacokinetics. Plasma samples (N = 4886) were collected from 487 patients with various solid tumors (mainly melanoma) in three clinical studies (MEK4592g, NO25395, GO28141). Cobimetinib was administered orally, once daily on either a 21-day-on/7-day-off, 14-day-on/14-day-off or 28-day-on schedule in a 28-day dosing cycle as single agent or in combination with vemurafenib. Cobimetinib doses ranged from 2.1 to 125 mg. NONMEM was used for pharmacokinetic analysis. A linear two-compartment model with first-order absorption, lag time and first-order elimination described cobimetinib pharmacokinetics. The typical estimates (inter-individual variability) of apparent clearance (CL/F), central volume of distribution (V2/F) and terminal half-life were 322 L/day (58 %), 511 L (49 %) and 2.2 days, respectively. Inter-occasion variability on relative bioavailability was estimated at 46 %. CL/F decreased with age. V2/F increased with body weight (BWT). However, the impact of age and BWT on cobimetinib steady-state exposure (peak and trough concentrations and AUC following the recommended daily dose of 60 mg 21-day-on/7-day-off) was limited (<25 % changes across the distribution of age and BWT). No significant difference in cobimetinib pharmacokinetics or steady-state exposure was observed between patient subgroups based on sex, renal function, ECOG score, hepatic function tests, race, region, cancer type, and co-administration of moderate and weak CYP3A inducers or inhibitors and vemurafenib. A population pharmacokinetic model was developed for cobimetinib in cancer patients. Covariates had minimal impact on steady-state exposure, suggesting no need for dose adjustments and supporting the recommended dose for all patients.


Dyspnea lusoria: compression of the pulmonary artery by a Kommerell's diverticulum.
We report a rare case of Kommerell's diverticulum presenting as dyspnea on exertion. After careful physical examination, selective imaging tests demonstrated a significant flow abnormality in the left pulmonary artery caused by an aneurysm of the descending aorta in the area of the ligamentum arteriosum.


[Intracerebral bleeding as the first symptom of a congenital anaplastic astrocytoma].
Anaplastic astrocytomas in neonates are extremely rare. Newborns, however, often have neuroectodermal central nervous tumours. We report about a female term newborn (birth weight 3,600 g, APGAR 9/10/10), who had shrill screams, intermittent shivering and bradycardia. An ultrasound scan of the brain showed an intracerebral bleeding. Therefore, the child was transferred to the intensive care unit of our hospital. A CT scan showed cerebral bleeding in the left parieto-occipital region, partially clotted, with a space-demanding effect. The intracerebral bleeding in the left occipital region was cleared out. No tumour was found, but an anaplastic astrocytoma (WHO Grade III) was diagnosed histologically. Serial ultrasound investigations of the brain showed a normal midline and a redevelopment of the left-sided ventricle. After surgery no tumour was visible in the MRI. Six weeks later, a tumour was found in the area of the initial bleeding region on MRI. Congenital anaplastic astrocytomas have a variable outcome, with different survival rates as compared to adults. In the literature, survival rates of 36-50 % were found after complete tumour resection. In cases of neonatal intracerebral bleeding, a tumour might be the cause of the haemorrhage.


Genetic and environmental influences on applied creativity: A reared-apart twin study.
Applied creativity involves bringing innovation to real-life activities. The first reared-apart twin study assessing genetic and environmental origins of applied creativity, via Draw-a-House (DAH) and Draw-a-Person (DAP) tasks, is presented. Participants included 69 MZA and 53 DZA twin pairs from the Minnesota Study of Twins Reared Apart. Drawings were evaluated by four artists and four non-artists. Genetic effects were demonstrated for the DAP (.38-.47), but not for the DAH. Creative personality showed genetic effects (.50), and modest, but significant correlations with scores on the two drawings (rs = .17-.26). Both genetic and nonshared environmental influences underlie variance in applied creativity. Individuals concerned with enhancing creativity among students and others may better understand individual differences in performance and training.


Gastrointestinal dysfunction and its effects on nutrition in CF.
Prevention of malnutrition, a consequence of elevated energy requirements, increased losses and low caloric intake, is one of the main goals in the treatment of cystic fibrosis. Caloric stool losses, catch-up growth and an elevated energy expenditure, even in the absence of overt lung disease and malabsorption, have led to recommendations for a caloric intake of 120-150% of the recommended daily allowances. A high energy intake with a fat content of at least 40 calorie % and adequate pancreatic supplementation has shown to improve growth and median age of survival. As a rational treatment of the fundamental disturbance in cystic fibrosis, a decrease in chloride permeability across epithelia, is not yet available, treatment should be concentrated on adequate nutritional support in combination with optimal correction of those gastrointestinal abnormalities including faecal bile acid loss, small intestinal abnormalities, pancreatic insufficiency, hormonal abnormalities and disturbances in gastrointestinal motility, which may aggravate maldigestion and malabsorption.


The migraine syndrome in children and adolescents.
Headache has a remarkably constant occurrence in different socioeconomic, cultural, geographic, and ethnic groups. Between 70% and 90% of people surveyed had at least one headache in the preceding year. Twenty-one percent of children have had significant tension or migraine headaches by 15 years of age, and an additional 54% have experienced occasional, less significant headache. Of children who had migraine, 42% lost time from school because of their headaches. Three percent of adults lost time from work because of headaches. This common symptom requires an organized approach to diagnosis and treatment.


Monitoring response to primary chemotherapy in breast cancer using dynamic contrast-enhanced magnetic resonance imaging.
Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) allows analysis of both tumor volume and contrast enhancement pattern using a single tool. We sought to investigate whether DCE-MRI could be used to predict histological response in patients undergoing primary chemotherapy (PCT) for breast cancer. Thirty patients with breast cancer, clinical diameter > 3 cm or stage III A/B, received anthracycline and taxane based PCT. DCE-MRI was performed at the baseline, after two cycles and after four cycles of PCT, before surgery. Histological response was assessed using a five-point scheme. Grade 4 (small cluster of dispersed residual cancer cells) and grade 5 (no residual viable cancer cell) were defined as a major histopathological response (MHR). Univariate analysis showed that a > 65% reduction in the tumor volume and a reduction in the early enhancement ratio (ECU) after two cycles of PCT were associated with a MHR. Multivariate analysis revealed that tumor volume reduction after two cycles of PCT was independently associated with a MHR (odds ratio [OR] 39.968, 95% confidence interval [CI] 3.438-464.962, p < 0.01). ECU reduction was still associated with a MHR (OR 2.50, 95% CI 0.263-23.775), but it did not retain statistical significance (p = 0.42). Combining tumor volume and ECU reduction after two cycles of PCT yielded a 93% diagnostic accuracy in identifying tumors achieving a pathological complete response (pCR) (histopathological grade 5). DCE-MRI allows prediction of the effect of neoadjuvant chemotherapy in breast cancer. Although in our study tumor volume reduction after two cycles had the strongest predictive value, DCE-MRI has the potential to provide functional parameters that could be integrated to optimize neoadjuvant chemotherapy strategies.


In vivo irreversible and reversible transverse relaxation rates in human cerebral cortex via line scans at 7 T with 250 micron resolution perpendicular to the cortical surface.
Understanding how and why MR signals and their associated relaxation rates vary with cortical depth could ultimately enable the noninvasive investigation of the laminar architecture of cerebral cortex in the living human brain. However, cortical gray matter is typically only a few millimeters thick, making it challenging to sample many cortical depths with the voxel sizes commonly used in MRI studies. Line-scan techniques provide a way to overcome this challenge and here we implemented a novel line-scan GESSE pulse sequence that allowed us to measure irreversible and reversible transverse relaxation rates-R2 and R2´, respectively-with extremely high resolution (250 μm) in the radial direction, perpendicular to the cortical surface. Eight healthy human subjects were scanned at 7 T using this sequence, with primary visual cortex (V1) targeted in three subjects and primary motor (M1) and somatosensory cortex (S1) targeted in the other five. In all three cortical areas, a peak in R2 values near the central depths was seen consistently across subjects-an observation that has not been made before, to our knowledge. On the other hand, no consistent pattern was apparent for R2´ values as a function of cortical depth. The intracortical R2 peak reported here is unlikely to be explained by myelin content or by deoxyhemoglobin in the microvasculature; however, this peak is in accord with the laminar distribution of non-heme iron in these cortical areas, known from prior histology studies. Obtaining information about tissue microstructure via measurements of transverse relaxation (and other quantitative MR contrast mechanisms) at the extremely high radial resolutions achievable through the use of line-scan techniques could therefore bring us closer to being able to perform "in vivo histology" of the cerebral cortex.


Cyclic Testing of the 6-Strand Tang and Modified Lim-Tsai Flexor Tendon Repair Techniques.
In this study, we compared the Tang repair technique with the 6-strand modified Lim-Tsai repair technique under cyclic testing conditions. Twenty fresh-frozen porcine flexor tendons were randomized into 2 groups for repair with either the modified Lim-Tsai or the Tang technique using Supramid 4-0 core sutures and Ethilon 6-0 epitendinous running suture. The repaired tendons were subjected to 2 stage cyclic loading. The survival rate and gap formation at the repair site were recorded. Tendons repaired by the Tang technique achieved an 80% survival rate. None of the modified Lim-Tsai repairs survived. The mean gap formed at the end of 1000 cycles was 1.09 mm in the Tang repairs compared with 4.15 mm in the modified Lim-Tsai repairs. The Tang repair is biomechanically stronger than the modified Lim-Tsai repair under cyclic loading. The Tang repair technique may exhibit a higher tolerance for active mobilization after surgery with less propensity for gap formation.


Five-year outcomes of endosuture aneurysm repair in patients with short neck abdominal aortic aneurysm from the ANCHOR registry.
Hostile aortic neck anatomies such as proximal short necks are known to put patients at an increased risk for type IA endoleaks, migration, and need for reinterventions. The Heli-FX EndoAnchor System was designed to improve seal of aortic stent grafts. Endosuture aneurysm repair (ESAR) using EndoAnchors with the Endurant stent graft has been shown to be safe and effective for the treatment of patients with short necks through one year. This study reports the 5-year patient outcomes of the Aneurysm Treatment using the Heli-FX EndoAnchor System Global Registry (ANCHOR) short neck regulatory cohort. The 70 patients from the ANCHOR Registry were cohort submitted to regulators for approval of the Endurant short neck indication. Patients had an infrarenal neck length of ≥ 4 mm and <10 mm. At 5 years, this short neck cohort had clinical and imaging follow-up compliance rates of 85% (28/33) and 70% (23/33), respectively. The short neck cohort had a mean age of 71.3±8.1 years and was 27.1% (19/70) female. Kaplan Meier freedom from all-cause mortality was 68.5 ± 6.2%, freedom from aneurysm-related mortality was 90.1 ± 4.5%, freedom from any endovascular or surgical secondary procedure was 76.9 ± 7.2%, and freedom from rupture was 95.6 ± 3.2%. Eight patients had a total of nine type IA endoleaks detected through 5 years, of which three resolved spontaneously by the next follow-up visit. There were two patients with renal complications who did not undergo reintervention and there were no device migrations reported through 5 years. After 5 years, 68.2% of patients (15/22) had sac regression, 13.6% (3/22) had stable sacs, and 18.2% (4/22) had increased sac diameter as compared with their 1-month measurements. After ESAR treatment using Heli-FX EndoAnchors with Endurant, the 5-year outcomes of the short neck cohort from the ANCHOR registry had encouraging results with regards to proximal neck-related complications, secondary procedures, and sac regression. This review of ESAR in patients with short proximal necks showed positive outcomes through 5 years although follow-up of a larger cohort is necessary.


The role of allergy in manifestations of respiratory disease in adult cystic fibrosis.
Variability is present in the expression of the clinical phenotype in cystic fibrosis (CF). Part of this variability may be explained by the coexistence of allergy in CF. To determine the rate of allergy in adult CF and evaluate the association between allergy and the manifestations of upper and lower airway disease. We performed a cross-sectional study of consecutive patients enrolled in a university hospital adult CF clinic. Allergen specific IgE was determined by radioallergosorbent and skin prick tests to common aeroallergens. We characterized features of upper and lower airway disease by clinical evaluation of rhinitis and spirometry before allergy testing. The study population consisted of 55 patients. Allergen specific IgE was present to at least 1 aeroallergen in 67% by skin prick testing and 80% by radioallergosorbent testing. Rhinitis occurred in 50% of the population and was associated with immediate-type hypersensitivity to aeroallergens other than molds. The frequency of rhinitis increased when there was sensitization to a greater number of aeroallergens and rarely occurred in the absence of allergic sensitization. There was no detectable difference in lung function between those with and without allergic sensitization. Immediate-type hypersensitivity to aeroallergens commonly occurs in adult CF. The coexistence of allergy in CF is associated with clinical features of rhinitis. Because allergic manifestations of CF warrant appropriate therapy, individuals with CF should be evaluated for coexistent allergy.


The effect of hyperthermia on intracellular K+ in Chinese hamster ovary cells.
The effect of hyperthermia on intracellular K+ concentrations was studied in Chinese hamster ovary (CHO) cells in vitro, using a flame photometer. Intracellular K+ concentrations decreased with increasing exposure time at temperatures from 40 degrees C to 45 degrees C. The decrease in K+ concentrations preceded any loss of reproductive capability at 43 degrees C and also occurred at the non-lethal temperature of 40 degrees C. Prolonged exposure to 45 degrees C resulted in an irreversible decrease in K+ concentrations. The decrease in K+ concentrations at elevated temperatures was not accounted for by changes in cell volume, loss of cells or failure of the Na+/K+ pump.


Molecular mechanism of long-term neuroprotective effects of gradual flow restoration on cerebral ischemia reperfusion injury in MCAO rats.
Ischemia-reperfusion injuries (IRIs) can aggravate the condition of some patients with acute occlusion of major intracranial artery (AOMIA) who received endovascular thrombectomy. Here, we provided data confirming the association of Repressor Element-1 Silencing Transcription factor (REST) with the long-term neuroprotective effect of the middle cerebral artery occlusion (MCAO) rats underwent Gradual Flow Restoration (GFR). Long term neuroprotective effects of GFR intervention were evaluated on MCAO rats model after 3d and 7d reperfusion. The neurological deficit score and TTC staining were performed to evaluate the degree of brain damage in GFR and other interventions at different time. Differentially expressed genes related to cerebral ischemia reperfusion injury (CIRI) were initially screened and identified using GSE32529 microarray analysis. REST protein expression in rat brain cortex infarction was detected by Western blot analysis. MCAO rats intervened with GFR exhibited reduced neurological deficit (P < 0.05) and alleviated brain infarction volume (P < 0.01). The REST gene with up-regulated expression and its downstream genes with down-regulated expression were screened by Microarray analysis. The brain cortex infarction in MCAO rats produced high levels of REST expression. The GFR intervention inhibited REST expression, and alleviated brain injury on MCAO rats. Our results demonstrated that GFR intervention plays a long-term neuroprotective role and reduces brain edema and damage at reperfusion, possibly by inhibiting REST expression.


RNA In Situ Hybridization on Plant Tissue Sections: Expression Analysis at Cellular Resolution.
RNA in situ hybridization offers a means to study the spatial expression of candidate genes by making use of specific, labelled RNA probes on thin tissue sections. Unlike other methods, such as promoter GUS fusions, for which all regulatory sequences should be available and transgenic plants have to be generated, RNA in situ hybridization allows specific and direct detection of even low abundant transcripts at cellular resolution. Although various protocols exist, the results published throughout the literature indicate a very obvious problem of the technique: each step has the potential to affect the outcome, that is, the signal strength, presence or absence of background, and visibility of individual cells. The protocol described here tries to avoid all these problems by addressing each step in detail and providing advice regarding critical steps for a distinct visualization of gene expression on intact tissue sections without any background.


PD0325901, an ERK inhibitor, enhances the efficacy of PD-1 inhibitor in non-small cell lung carcinoma.
ERK pathway regulated the programmed death ligand-1 (PD-L1) expression which was linked to the response of programmed death-1 (PD-1)/PD-L1 blockade therapy. So it is deducible that ERK inhibitor could enhance the efficacy of PD-1 inhibitor in cancer immunotherapy. In this study, PD0325901, an oral potent ERK inhibitor, strongly enhanced the efficacy of PD-1 antibody in vitro and in vivo models in non-small cell lung carcinoma (NSCLC) cells. Mechanistically, PD0325901 or shRNA-ERK1/2 significantly downregulated the PD-L1 expression in NSCLC cells and increased the CD3+ T cells infiltration and functions in tumor tissue. There was a positive correlation between the p-ERK1/2 expression and PD-L1 expression in patients with NSCLC. And the patients with low p-ERK1/2 expression were observed a high response rate of PD-1/PD-L1 blockage therapy. Our results demonstrate that PD0325901, an ERK inhibitor, can enhance the efficacy of PD-1 blockage against NSCLC in vitro and in vivo models. And the combination of ERK inhibitor such as PD0325901 and PD-1/PD-L1 blockage is a promising regimen and encouraged to be further confirmed in the treatment of patients with NSCLC.


Mechanism of action of the monosialoganglioside GM1 as a modulator of CD4 expression. Evidence that GM1-CD4 interaction triggers dissociation of p56lck from CD4, and CD4 internalization and degradation.
Analyzing the mechanisms underlying the capability of the monosialoganglioside GM1 to induce CD4 modulation we observed that GM1 has a dual effect on the CD4 molecule. GM1 treatment of the lymphoma cell line MOLT-3 and CD4-transfected HeLa cells for times shorter than 30 min prevented binding of monoclonal antibodies (mAbs) recognizing epitopes located within the first NH2-terminal domains of CD4, but not of the OKT4 mAb, which binds to the region of CD4 proximal to the transmembrane domain. However, no binding of the OKT4 mAb was observed after a few hours of treatment with GM1 in both MOLT-3 cells and HeLa cells transfected with an intact CD4 molecule, but not in HeLa cells transfected with a CD4 molecule lacking the bulk of the cytoplasmic domain, suggesting that modulation of CD4 by GM1 depends on the integrity of the cytoplasmic domain. GM1 treatment blocked binding of several mAbs which recognize epitopes located within the first two NH2-terminal domains of CD4 and did not induce CD4 down-modulation if MOLT-3 cells were preincubated with the OKT4A or the OKT4 mAbs. Immunoprecipitation studies with [35S]methionine-labeled MOLT-3 cells showed that GM1-induced CD4 down-modulation was accompanied by CD4 degradation, and this was preceded by dissociation of p56lck from CD4. GM1-induced CD4 down-modulation, dissociation of p56lck from CD4, and CD4 degradation were unaffected by staurosporine, which, on the contrary, blocked these events in response to phorbol 12-myristate 13-acetate. These observations demonstrate that the first action of GM1 is to mask epitopes located within the first two NH2-terminal domains; then, GM1 triggers protein kinase C-independent signals which cause p56lck dissociation from CD4 and the delivery of the molecule to an intracellular compartment where it is eventually degraded.