Advanced MR imaging of acute stroke: the University of California at Los Angeles endovascular therapy experience.
Rapid implementation of multimodal MR imaging, including diffusion-perfusion imaging before and following endovascular therapies for treatment of acute ischemic stroke, has yielded novel observations and expanded contemporary knowledge of acute stroke pathophysiology. For more than a decade at the University of California at Los Angeles, an intensive imaging strategy has been used to establish stroke diagnosis, delineate early patterns of cerebral ischemia, and monitor response to various reperfusion techniques. Angiographic findings before thrombolysis have substantiated stroke subtype, and documentation of concomitant recanalization has provided considerable insight regarding the vascular and imaging correlates of intra-arterial therapies. MR imaging studies have progressively characterized the complex pathophysiology of acute ischemic stroke, providing imaging strategies and models that may be used to optimize acute stroke care. The ensuing review emphasizes salient aspects of these neuroimaging studies, addressing numerous facets of acute stroke pathophysiology.


Oral administration of liposome-entrapped cysteamine and the distribution pattern in blood, liver and spleen.
Oral administration of liposome-entrapped cysteamine induces an increase in the concentration of exogenous sulphur compounds in blood (plasma), liver and spleen. Among those sulphur compounds, an important amount of plasma thiols can be related to a protection of cysteamine in the digestive tract. This can account for the radioprotective effect of a liposomal-cysteamine suspension in rodents, and clearly demonstrates the interest of such a preparation in radioprotection.


[Study on pulmonary delivery system of salmon calcitonin in rats].
To study the serum calcitonin(CT) levels in vivo after pulmonary delivery to rats and the effects of absorption enhancers. Salmon calcitonin (sCT) solution was delivered by intramuscular injection or intratracheal administration to rats. The absorption of sCT was determined by radioimmunoassay method. Changes in serum drug levels after the pulmonary administration of sCT with various absorption enhancers or in different pH conditions were investigated. Compared with intramuscular administration, the relative bioavailability of sCT solution (2 micrograms.kg-1) given by intratracheal route was 121.0%. Transpulmonary absorption of sCT solution, with some absorption enhancers (Brij78, sodium cholate, lecithin, chitosan and sodium caprylate) or under acidic conditions (pH3.0), was significantly greater than the values obtained in controls. sCT solution showed higher bioavailability after pulmonary administration.


Fitness cost, gyrB mutation, and absence of phosphotransferase system fructose specific IIABC component in novobiocin-resistant Streptococcus iniae vaccine strain ISNO.
To understand the fitness cost of novobiocin-resistance in an attenuated Streptococcus iniae vaccine strain ISNO compared to its virulent parent strain ISET0901, cell proliferation rate of the two strains were compared to each other. Our results revealed that the cell proliferation rates of ISNO were significantly (P<0.05) smaller than that of ISET0901. To understand whether there was any mutation at the target site of novobiocin, DNA gyrase subunit B (gyrB) was sequenced from both strains. Sequencing results revealed a point mutation of AGA to AGC, resulting in a deduced amino acid substitution of R635S. To determine whether any unique DNA sequence was present in ISET0901 but absent in ISNO, PCR-select bacterial genome subtractive hybridization was performed. A phosphotransferase system fructose specific IIABC component sequence was confirmed to be present in ISET0901 but absent in ISNO. Using genomic DNAs from ten field-strains of S. iniae as templates, the phosphotransferase system fructose specific IIABC component sequence was found to be present in five highly virulent strains, but absent in five avirulent strains. Taken together, our results suggest that: (1) As fitness cost of novobicin resistance, ISNO had significantly smaller cell proliferation rate; (2) point mutation at target site gyrB resulting in R635S substitution was associated with novobiocin resistance in ISNO; and (3) phosphotransferase system fructose specific IIABC component was associated with virulence of S. iniae.


Determining the angle of screw placement for internal fixation of calcaneal fractures.
This study was designed to determine the angle off the perpendicular to the lateral calcaneal wall at which a screw would need to be angled to avoid entering the subtalar joint medially. It also determined the distance inferior to the articular surface at which a screw would need to be placed to avoid that penetration if the screw was placed perpendicular to the lateral calcaneal wall. Eight specimens were dissected and the calcaneus was removed. The measurements were taken from portions of the subtalar joint. To avoid medial penetration, a screw at the anterior portion of the posterior facet would need to be angled 5 degrees plantar medially or placed 1.5 millimeters below the edge of the articular surface. In the midportion, placements are 20 degrees or 10 millimeters; posteriorly, placements are 32 degrees or 15 millimeters. This study shows that using a lateral approach, it is possible to position internal fixation screws without disrupting the articular surfaces of the medial subtalar joint.


A new combined process for efficient removal of Cu(II) organic complexes from wastewater: Fe(III) displacement/UV degradation/alkaline precipitation.
Efficient removal of heavy metals complexed with organic ligands from water is still an important but challenging task now. Herein, a novel combined process, i.e., Fe(III)-displacement/UV degradation/alkaline precipitation (abbreviated as Fe(III)/UV/OH) was developed to remove copper-organic complexes from synthetic solution and real electroplating effluent, and other processes including alkaline precipitation, Fe(III)/OH, UV/OH were employed for comparison. By using the Fe(III)/UV/OH process, some typical Cu(II) complexes, such as Cu(II)-ethylenediaminetetraacetic acid (EDTA), Cu(II)-nitrilotriacetic acid (NTA), Cu(II)-citrate, Cu(II)-tartrate, and Cu(II)-sorbate, each at 19.2 mg Cu/L initially, were efficiently removed from synthetic solution with the residual Cu below 1 mg/L. Simultaneously, 30-48% of total organic carbon was eliminated with exception of Cu(II)-sorbate. Comparatively, the efficiency of other processes was much lower than the Fe(III)/UV/OH process. With Cu(II)-citrate as the model complex, the optimal conditions for the combined process were obtained as: initial pH for Fe(III) displacement, 1.8-5.4; molar ratio of [Fe]/[Cu], 4:1; UV irradiation, 10 min; precipitation pH, 6.6-13. The mechanism responsible for the process involved the liberation of Cu(II) ions from organic complexes as a result of Fe(III) displacement, decarboxylation of Fe(III)-ligand complexes subjected to UV irradiation, and final coprecipitation of Cu(II) and Fe(II)/Fe(III) ions. Up to 338.1 mg/L of Cu(II) in the electroplating effluent could be efficiently removed by the process with the residual Cu(II) below 1 mg/L and the removal efficiency of ∼99.8%, whereas direct precipitation by using NaOH could only result in total Cu(II) removal of ∼8.6%. In addition, sunlight could take the place of UV to achieve similar removal efficiency with longer irradiation time (90 min).


Engineered knottin peptide enables noninvasive optical imaging of intracranial medulloblastoma.
Central nervous system tumors carry grave clinical prognoses due to limited effectiveness of surgical resection, radiation, and chemotherapy. Thus, improved strategies for brain tumor visualization and targeted treatment are critically needed. We demonstrate that mouse cerebellar medulloblastoma (MB) can be targeted and illuminated with a fluorescent, engineered cystine knot (knottin) peptide that binds with high affinity to αvβ3, αvβ5, and α5β1 integrin receptors. This integrin-binding knottin peptide, denoted EETI 2.5F, was evaluated as a molecular imaging probe in both orthotopic and genetic models of MB. Following tail vein injection, fluorescence arising from dye-conjugated EETI 2.5F was localized to the tumor compared with the normal surrounding brain tissue, as measured by optical imaging. The imaging signal intensity correlated with tumor volume. Due to its unique ability to bind to α5β1 integrin, EETI 2.5F showed superior in vivo and ex vivo brain tumor imaging contrast compared with other engineered integrin-binding knottin peptides and with c(RGDfK), a well-studied integrin-binding peptidomimetic. Next, EETI 2.5F was fused to an antibody fragment crystallizable (Fc) domain (EETI 2.5F-Fc) to determine if a larger integrin-binding protein could also target intracranial brain tumors. EETI 2.5F-Fc, conjugated to a fluorescent dye, illuminated MB following i.v. injection and was able to distribute throughout the tumor parenchyma. In contrast, brain tumor imaging signals were not detected in mice injected with EETI 2.5F proteins containing a scrambled integrin-binding sequence, demonstrating the importance of target specificity. These results highlight the potential of using EETI 2.5F and EETI 2.5-Fc as targeted molecular probes for brain tumor imaging.


The Systemic Immune-Inflammation Index May Predict the Coronary Slow Flow Better Than High-Sensitivity C-Reactive Protein in Patients Undergoing Elective Coronary Angiography.
A total of 197 patients (102 patients with CSF; 95 patients with normal coronary flow) were included in this retrospective study. Clinical and angiographic characteristics of patients were obtained from hospital records. Patients with CSF had higher SII, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte (PLR), and high-sensitivity C-reactive protein (hsCRP) levels compared with the control group. Body mass index (p=0.022, OR 1.151, 95% CI 1.121-1.299), low-density lipoprotein (p=0.018, OR 1.028, 95% CI 1.005-1.052), hsCRP (p=0.044, OR 1.161, 95% CI 1.004-1.343), and SII (p < 0.001, OR 1.015, 95% CI 1.003-1.026) were independent predictors of CSF in the multivariable analysis. The optimal cutoff value of SII in predicting CSF was >877 in ROC curve analysis (p < 0.001, AUC = 0.892, 95% CI 0.848-0.936). This cutoff value of SII predicted the CSF with a sensitivity of 71.5% and specificity of 92.4%. Spearman correlation analysis showed a positive correlation between the mean TFC value and PLR, NLR, hsCRP, and SII. SII may be used as a better indicator for the prediction of CSF than hsCRP.


When do Children with Autism Spectrum Disorder Take Common Ground into Account During Communication?
One feature of autism spectrum disorder (ASD) is a deficit in verbal reference production, that is, providing an appropriate amount of verbal information for the listener to refer to things, people, and events. However, very few studies have manipulated whether individuals with ASD can take a speaker's perspective to interpret verbal reference. A critical limitation of all interpretation studies is that comprehension of another's verbal reference required the participant to represent only the other's visual perspective. Yet, many everyday interpretations of verbal reference require knowledge of social perspective (i.e., a consideration of which experiences one has shared with which interlocutor). We investigated whether 22 5;0-7;11-year-old children with ASD and 22 well-matched typically developing (TD) children used social perspective to comprehend (Study 1) and produce (Study 2) verbal reference. Social perspective-taking was manipulated by having children collaboratively complete activities with one of two interlocutors such that for a given activity, one interlocutor was Knowledgeable and one was Naïve. Study 1 found no between-group differences for the interpretation of ambiguous references based on social perspective. In Study 2, when producing referring terms, the ASD group made modifications based on listener needs, but this effect was significantly stronger in the TD group. Overall, the findings suggest that high-functioning children with ASD know with which interlocutor they have previously shared a given experience and can take this information into account to steer verbal reference. Nonetheless, they show clear performance limitations in this regard relative to well-matched controls. Autism Res 2018, 11: 1366-1375. © 2018 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: No one had studied if young children with autism spectrum disorder (ASD) could take into account previous collaboration with particular conversation partners to drive how well they communicate with others. In both their language understanding and spoken language, we found that five to 7-year-olds with ASD were able to consider what they had previously shared with the conversation partner. However, they were impaired when compared to typically developing children in the degree to which they tailored their spoken language for a specific listener.


Clinical assessment by MRI for patients with stage II cervical carcinoma treated by radiation alone in multicenter analysis: are all patients with stage II disease suitable candidates for chemoradiotherapy?
From recent randomized trials, patients with Stage II cervical carcinoma are thought to be candidates for chemoradiotherapy. To refine the strategy for Stage II patients, we performed a retrospective multi-institutional analysis using MRI. From three institutions, 84 patients with Stage II cancer diagnosed by MRI were entered into the study. All patients received intracavitary brachytherapy with (n = 83) or without (n = 1) external beam radiotherapy. Uni- and multivariate analyses were performed to identify the prognostic factors for overall survival (OAS), disease-free survival (DFS), pelvic control (PC), and distant metastasis-free survival (DMFS). The 5-year DFS rate of patients with maximal tumor size (D(max)) > or =50 mm (46.2%) was significantly lower than that for patients with D(max) <50 mm (88.0%; p <0.0001). Large size or volume and lymph node swelling were also significant prognostic factors of OAS, DFS, PC, and DMFS. In the multivariate model, size or volume was a significant prognostic factor of OAS, DFS, PC, and DMFS, and lymph node swelling was a prognostic factor for DFS, PC, and DMFS. Using these two prognostic factors, patients were divided into 3 subgroups. The 5-year DFS rate of patients with risk 0 (D(max) <50 mm and negative lymph node swelling), 1 (D(max) > or =50 mm or positive lymph node swelling), and 2 (D(max) > or =50 mm and positive lymph nodes) was 93.2%, 53.3%, and 25.0%, respectively (p <0.0001). In this stage-limiting analysis, we clarified the stratification according to clinical risk with the aid of MRI. For patients with low-risk factors, especially for elderly patients, radiation alone would probability be a feasible option. In the future, a randomized trial using criteria with MRI would help to identify the optimal strategy for patients with Stage II disease.


The role of oral oil administration in displaying the chylous tubes and preventing chylous leakage in laparoscopic para-aortic lymphadenectomy.
This study is aimed to investigate the possibility of preoperative oral oil administration in displaying the chylous tubes and preventing chylous leakage in laparoscopic para-aortic lymphadenectomy. In this retrospective nonrandomized study, of the 30 patients with gynecological malignancies who had indications for laparoscopic para-aortic lymphadenectomy up to renal vessels, 15 were administered preoperative oral oil (oil a administration) (control group) at our hospital between September 2017 and June 2018. The chylous tube displaying rates, incidences of chylous leakage, and other perioperative data of the two groups were compared. Successful display of chylous tubes was observed in 93.3% (14/15) patients in the oil administration group. The chylous leakage was zero in the oil administration group, and 33.3% (5/15) in the control group. The postoperative drainage duration (4.1 ± 1.0 days vs 7.6 ± 1.4 days, P = 0.000), somatostatin application time (0 day vs 5.9 ± 0.8 days), and postoperative hospital stay (6.0 ± 2.3 days vs 9.1 ± 2.1 days, P = 0.001) were significantly shorter in the oil administration group. The total cost is lower in the oil administration group (4972.52 ± 80.54 dollars vs 6260.80 ± 484.47 dollars, P = 0.000). Preoperative oil administration is a feasible and effective method to display the chylous tubes and to prevent the chylous leakage in para-aortic lymphadenectomy.


Collateral Circulation Predicts the Size of the Infarct Core and the Proportion of Salvageable Penumbra in Hyperacute Ischemic Stroke Patients Treated with Intravenous Thrombolysis.
We studied the impact of collateral circulation on CT perfusion (CTP) parametric maps and the amount of salvaged brain tissue, the imaging and clinical outcome at 24 h and at 3 months in a retrospective acute (<3 h) stroke cohort (105 patients) with anterior circulation thrombus treated with intravenous thrombolysis. Baseline clinical and imaging information were collected and groups with different collateral scores (CS) were compared. Binary logistic regression analyses using good CS (CS ≥2) as the dependent variable were calculated. CTP Alberta Stroke Program Early CT Score (ASPECTS) was successfully assessed in 58 cases. Thirty patients displayed good CS. Poor CS were associated with more severe strokes according to National Institutes of Health Stroke Scale (NIHSS) at arrival (15 vs. 7, p = 0.005) and at 24 h (10 vs. 3, p = 0.003) after intravenous thrombolysis. Good CS were associated with a longer mean onset-to-treatment time (141 vs. 121 min, p = 0.009) and time to CTP (102 vs. 87 min, p = 0.047), better cerebral blood volume (CBV) ASPECTS (9 vs. 6, p < 0.001), better mean transit time (MTT) ASPECTS (6 vs. 3, p < 0.001), better noncontrast CT (NCCT) ASPECTS (10 vs. 8, p < 0.001) at arrival and with favorable clinical outcome at 3 months (modified Rankin Scale ≤2, p = 0.002). The fraction of penumbra that was salvageable at arrival and salvaged at 24 h was higher with better CS (p < 0.001 and p = 0.035, respectively). In multivariate analysis, time from the onset of symptoms to imaging (p = 0.037, OR 1.04 per minute, 95% CI 1.00-1.08) and CBV ASPECTS (p = 0.001, OR 2.11 per ASPECTS point, 95% CI 1.33-3.34) predicted good CS. In similar multivariable models, MTT ASPECTS (p = 0.04, OR 1.46 per ASPECTS point, 95% CI 1.02-2.10) and NCCT ASPECTS predicted good CS (p = 0.003, OR 4.38 per CT ASPECTS point, 95% CI 1.66-11.55) along with longer time from the onset of symptoms to imaging (p = 0.045, OR 1.03 per minute, 95% CI 1.00-1.06 and p = 0.02, OR 1.05 per minute, 95% CI 1.00-1.09, respectively). CBV ASPECTS had a larger area under the receiver operating characteristic curve for good CS (0.837) than NCCT ASPECTS (0.802) or MTT ASPECTS (0.752) at arrival. Favorable CBV ASPECTS, NCCT ASPECTS and MTT ASPECTS are associated with good CS along with more salvageable tissue and longer time from the onset of symptoms to imaging in ischemic stroke patients treated with intravenous thrombolysis.


Transplant coronary artery disease: Incidence, progression and interventional revascularization.
Allograft coronary artery disease (CAD) remains the main factor responsible for late graft loss. This analysis describes data on incidence and progression of allograft CAD at our institute, as well as our experience with coronary interventions in heart transplant recipients. Angiographic results of cardiac transplant patients undergoing coronary angiography were prospectively selected and analyzed. Angiographic outcome at follow-up were assessed for all coronary revascularizations in denovo lesions. Four hundred thirty-two coronary angiographies were performed in a total of 246 patients. Seventy-six patients (30.9%) showed angiographic evidence of CAD with %DS>50%, of which 48 patients revealed significant stenosis with %DS>70% (19.5%). Within the first 5 years after the transplantation, 10.1% show angiographic signs of a CAD; at the time of 10.1 years, 50% of all heart transplant patients have developed a CAD. Once a CAD with %DS between 50% and 60% has evolved, the disease shows fast progression. Coronary intervention was performed in 28 vessels at an average time of 9.5 years after heart transplantation. Follow-up angiography was available for 27 vessels (1 death before re-angiography) within a mean follow-up period of 19.3 months. Binary restenosis was found in 7 out of 27 vessels (25.9%). Comparison of the occurrence of total occlusion in vessels with %DS>70% which were not revascularized to the occurrence of MACE after successful revascularization revealed better long term results in the group of patients with coronary intervention (p=0.04). Whereas coronary artery disease is found in rare cases within the first 5 years after heart transplantation, the incidence grows in exponential manner after this period. Mid-term follow-up after coronary intervention exhibit restenosis-rates which are similar to the ones of other high risk patients. Comparison of coronary intervention versus conservative treatment in vessels with %DS>70% show significant better mid-term outcome in the interventional group.


The effects of lobbying on the FDA's recall classification.
The US Food and Drug Administration (FDA) regulates goods accounting for 20% of US consumers' total expenditure. The agency's potential susceptibility to corporate lobbying and political influence may adversely affect the its abilities to fulfill its duties as a vital federal agency. This study assesses whether the FDA's product recall classifications in recall scenarios are influenced by firms' lobbying activities. The universe of all FDA recalls between 2012 and 2019 is obtained from the FDA's website. Firm names are matched to federal-level lobbying data obtained from the Center for Responsive Politics - a non-profit and nonpartisan organization that tracks lobbying expenditures and campaign contributions. Analyses are conducted using ordinary-least-squares regressions, in which the dependent variable is recall classification and independent variables are three different measures of firms' lobbying activities in the one year prior to the recall. Firms that engage in lobbying appear more likely to receive favourable classifications from the FDA. When examining the above results by product type, we find that classification of food recalls seems to be subject to lobbying influence, but the same does not appear to be true for drug and device recalls. Evidence is consistent with the conjecture that the distinction between medical and food firms may be a result of medical firms targeting lobbying efforts at FDA approvals, rather than recalls. Between 2012 and 2019, the FDA's product recall classifications seem to be significantly influenced by firms' lobbying activities. Lobbying firms appear to have received more favorable (i.e., less severe) recall classifications compared to non-lobbying firms.


Ghrelin Promotes Cortical Neurites Growth in Late Stage After Oxygen-Glucose Deprivation/Reperfusion Injury.
Acyl ghrelin, a novel brain-gut peptide, is an endogenous ligand for the growth hormone secretagogue receptor. Accumulated research data have shown that acyl ghrelin exercises a significant neuroprotective effect against cerebral ischemia/reperfusion (I/R) injury in animal models and in cultured neurons during the acute phase, usually, 1 day after cerebral ischemia. The chronic effects of acyl ghrelin 1 week after brain ischemia remain largely unknown. In this study, we explored the effects of acyl ghrelin on cultured organotypic brain slices and cortical neurons which were injured by oxygen-glucose deprivation/reperfusion(OGD/R) for 7 days. The underlying molecular mechanisms were deciphered based on label-free proteomic analysis. Acyl ghrelin treatment promoted neurite (axons and dendrites) growth and alleviated the neuronal damage in both cultured brain slices and neurons. Proteomic analysis showed that cell division control protein 42 (Cdc42) mediates the effects of acyl ghrelin on neurite growth. Acyl ghrelin stimulated the expression of Cdc42 and neurite growth in cultured neurons comparing with OGD/R group. Inhibition of Cdc42 attenuated the effects of acyl ghrelin. These results suggest that acyl ghrelin promotes neurite growth during the later stage after OGD/R injury via Cdc42. Our study suggests that acyl ghrelin may be promising to restore the neuronal structure in the late phase after stroke.


Regional differences in renewal rates of fibroblasts in young adult female mice.
Young adult female mice were given a total of 90 intraperitoneal injections each of (methyl-3H)thymidine (3HTdR) at intervals of 8 h over 30 days to establish renewal rates of fibroblasts in various locations. Radioautographs prepared from punch biopsy material of the ears, taken repeatedly during the labeling procedure, revealed an approximately linear increase of labeling indices of dermal fibroblasts with time. Labeling indices of fibroblasts at the end of repetitive injections of 3HTdR differed depending on the site and/or type of connective tissues examined. Low values were obtained for fibroblasts in the leptomeninx (3.9%), the tracheal wall (4.4%), the achilles tendon (5.8%) and the dermis of the ear (6.5%), while higher values were registered for fibroblasts located in the lower half of the abdominal dermis (12.3%), peritendinous sheaths (12.6%), the interstitial connective tissue of the thigh muscles (12.9%), the submucosa of the colon (23%), the fibrous capsule of the adrenals (25.7%), and the upper half of the abdominal dermis (26%). These regional differences, with the exception of the skin and possibly of the tracheal wall, did not correlate with local temperature. Possible additional factors influencing fibroblast renewal rates may include the type of connective tissue, the degree of vascularization, mechanical stress and hormonal action. Estimated turnover times, based on the assumptions of a DNA synthesis time of 6 h or more and a linear increase of labeling indices as a function of time of repetitive labeling, range from about 90 to more than 700 days. The higher values approximate the median life span of the mouse strain used.


On the identification of low allele frequency mosaic mutations in the brains of Alzheimer's disease patients.
The cause of sporadic Alzheimer's disease (AD) remains unclear. Given the growing evidence that protein aggregates can spread in a "prion-like" fashion, we reasoned that a small population of brain cells producing such "prion-like" particles due to a postzygotic acquired mutation would be sufficient to trigger the disease. Deep DNA sequencing technology should in principle allow the detection of such mosaics. To detect the somatic mutations of genes causing AD present in a small number of cells, we developed a targeted deep sequencing approach to scrutinize the genomic loci of APP, PSEN1, and PSEN2 genes in DNA extracted from the entorhinal cortex, one of the brain regions showing the earliest signs of AD pathology. We also included the analysis of the MAPT gene because mutations may promote tangle formation. We validated candidate mutations with an independent targeted ultradeep amplicon sequencing technique. We demonstrate that our approach can detect single-nucleotide mosaic variants with a 1% allele frequency and copy number mosaic variants present in as few as 10% of cells. We screened 72 AD and 58 control brain samples and identified three mosaic variants with low allelic frequency (∼1%): two novel MAPT variants in sporadic AD patients and a known PSEN2 variant in a Braak II control subject. Moreover, we detected both novel and known pathogenic nonmosaic heterozygous variants in PSEN1 and PSEN2 in this cohort of sporadic AD patients. Our results show that mosaic mutations with low allelic frequencies in AD-relevant genes can be detected in brain-derived DNA, but larger samples need to be investigated before a more definitive conclusion with regard to the pathogenicity of such mosaics can be made.


Cloning and expression of dnaK gene from Bacillus pumilus of hot water spring origin.
A set of thermotolerant strains isolated from hot springs of Manikaran and Bakreshwar (India) were selected with an aim to isolate dnak gene which encodes DnaK protein. The gene dnaK along with its flanking region was successfully amplified from 5 different strains (4 from Bakreshwar and one from Manikaran). Restriction fragment length polymorphism (RFLP) revealed that amplicons were almost identical in sequence. The dnak gene from one representative, Bacillus pumilus strain B3 isolated from Bakreshwar hot springs was successfully cloned and sequenced. The dnaK gene was flanked by gene grpE on one side. The dnaK gene was 1842 bp in length encoding a polypeptide of 613 amino acid residues. Calculated molecular weight and pI of the protein were 66,128.36 Da and 4.72 respectively. The deduced amino acid sequence of this gene shared high sequence homology with other DnaK proteins and its homologue Hsp 70 from other microorganisms, but possessed 36 substitutions and two insertions, as compared to DnaK protein of Bacillus subtilis. The dnaK gene of B. pumilus was successfully expressed in Escherichia coli BL 21 (DE3) using pET expression systems. Heterologous expression of dnaK of B. pumilus in E. coli BL 21 (DE3) allowed for the growth of E. coli up to 50 °C and survival up to 60 °C for 16 h, suggesting that dnak from B. pumilus imparts tolerance to host cells under high temperature. This novel gene can be an important component for possible utilization in abiotic stress management of plants.


Biomagnetic activity in ovarian lesions.
BACKGROUND-MATERIALS: This study aimed to investigate biomagnetic activity in benign and malignant ovarian diseases using the biomagnetometer SQUID. Magnetic recordings were obtained from 40 patients with palpable ovarian lesions. 19 of these were invasive carcinomas, and 21 were benign ovarian lesions. We used a one channel biomagnetometer SQUID (superconducting quantum interference device), in order to measure the magnetic field from benign and malignant ovarian diseases. Interestingly, the ovarian lesion waveforms and the corresponding spectral densities were of high amplitude in most (96%) malignant ovarian lesions, and of low amplitude in most (95%) benign ovarian diseases. These findings were of statistical significance (students t-test p < 0.005). It is suggested that biomagnetic measurement of benign and malignant ovarian diseases, which is an entirely new application of SQUID technology, is a promising procedure for assessing ovarian tumors.


Airflow grades, outcome measures and response to pulmonary rehabilitation in individuals after an exacerbation of severe chronic obstructive pulmonary disease.
Individuals with COPD may be staged according to symptoms and exacerbation history (GOLD groups: A-D) and on airflow obstruction (GOLD grades: 1-4). Guidelines recommend pulmonary rehabilitation (PR) for these individuals, including those recovering from an exacerbation (ECOPD) OBJECTIVE: To evaluate whether in individuals with clinically severe COPD, recovering from an ECOPD, the effect size of an in-hospital PR program would be affected by airflow severity grades and assessed outcome measures. Retrospective, multicentre study. Participants were compared according to different GOLD airflow grades. In addition to the MRC dyspnoea scale, six-minute walking distance test and COPD assessment test (CAT), Barthel dyspnoea index (Bid), and Short Physical Performance Battery (SPPB) were assessed, evaluating the proportion of individuals reaching the minimum clinically important difference (MCID) (responders). Data of 479 individuals, completing the program were evaluated. Most of the participants were allocated in GOLD grades 4, (57.6%) and 3 (22.1%). All outcome measures significantly improved after PR (p < 0.05), without any significant difference in the proportion of responders in any measure. in individuals with severe COPD, recovering from ECOPD the success rate of PR does not depend on airflow severity, or outcome measure assessed. In addition to the most used outcome measures, also Bid and SPPB are sensitive to PR.


A randomized trial comparing triple-drug and double-drug therapy in renal transplantation.
A controlled trial was carried out in 86 cadaveric and 14 living haploidentical renal transplant recipients to compare the effects of low doses of cyclosporine (CsA), azathioprine (Aza) and steroids with those of higher doses of CsA plus steroids. Patients were followed for 12-26 months after transplantation. The actuarial 2-year patient and graft survival rate was 100% for living-donor transplants. In cadaver renal transplants the 2-year patient survival rate was 100% for patients assigned to the triple regimen and 93% for those allocated to the double regimen. The actuarial 2-year cadaver graft survival rates were 86% and 90.6%, respectively. There were significantly more patients who had severe infections (P less than 0.05), particularly interstitial pneumonia (P less than 0.005), in the double-therapy group. On the other hand, there were more patients who rejected and more patients with severe rejections; more pulses of steroids were also required for patients on the triple regimen, although these differences were not significant. The mean trough blood levels of cyclosporine at the various times were about half as high in patients on triple therapy. There were no differences between the two groups in creatinine clearance at any time. A control renal biopsy, taken from patients with stable renal function after 6-12 months, showed only mild abnormalities. The lesions were semiquantitatively assessed. There was a higher score for interstitial infiltrate in patients on triple therapy (P less than 0.05). On the other hand, the incidence and the mean score of interstitial fibrosis were greater in patients on double therapy, although these differences were not significant. Thus, although similar results were obtained with both regimens, at the doses we used double therapy seems to have more powerful immunosuppressive effects and may prevent rejection, either acute or chronic, better. However, it might expose the patient to a greater risk of infection and of cyclosporine-related nephrotoxicity than triple therapy.


Development and validation of psychosocial problems scale for type 2 diabetes (PPSTD).
To develop and validate the Psychosocial Problems Scale for type 2 diabetics. The study was conducted at Lahore from November 2017 to July 2018 in three phases. The first phase related to item generation in which endorsements was taken from experts. In the second phase, a pilot study was conducted and in the third phase, psychometric properties were established. In the third phase, the scale was administered on a sample of diabetics aged 30-60 years. Exploratory factor analysis was used to establish construct validity of the scale. Correlation was carried out to establish concurrent validity of the scale with Depression Anxiety and Stress Scale and Psychosocial Problems Scale for type 2 diabetics. The mean age of 395 subjects was 47.19±8.76 years. Three factors that emerged from factor analysis were social/occupational, sexual life satisfaction and psychological. Significant positive correlation (p<0.001) was found with both the scales. To establish the discriminant validity, correlation was computed and significant negative correlation (p<0.01) was found between Satisfaction with Life Scale and Psychosocial Problems Scale for type 2 diabetics (p<0.05). The psychosocial problem scale for type 2 diabetics was found to be a valid and reliable tool to identify psychosocial problems of patients with type 2 diabetes.


Comment on "Stability of Tsallis entropy and instabilities of Rényi and normalized Tsallis entropies: a basis for q-exponential distributions".
It is shown that the Rényi entropy is as stable as the Tsallis entropy at least for the Abe-Lesche counter examples.


First approaches to the depuration process of trace metals in the burrowing crab Neohelice granulata from a temperate wetland in South America: Bahía Blanca estuary, Argentina.
Trace metal contamination is among the major concerns of stakeholders due to its potential adverse effects on biota and humans, even at low concentrations. Few studies have recently focused on the ability of organisms to depurate trace metals from different tissues. Therefore, we carried out this study to evaluate the bioconcentration of trace metals (Cd, Zn, Pb, Ni, Mn, Fe, Cr, Cu) and the depuration process of these pollutants in two tissues (soft tissue and carapace) in an estuarine benthic crab model, Neohelice granulata. The results indicate that Cu and Zn were the highest bioconcentrated metals in crab tissues, while other metals, such as Pb and Cr, were found in sediments but were not bioconcentrated. On the other hand, Cd was found in crabs but not in sediments. The depuration indicates a total decline in Ni and a significant decrease in Cu and Fe in the soft tissues after the experiment. However, the concentration of the trace metals in the carapace before and after the depuration did not show any significant variation except in the Mn, in which the levels decreased significantly at the end of the depuration. Thus, we recommend continuing to explore metal detoxification in bioindicator species, such as N. granulata, in order to understand the efficiency of the mechanisms of depuration of trace metals.


Septum Resection Prior to In Vitro Fertilization-Embryo Transfer: A Retrospective Controlled Study.
To investigate the effectiveness of surgery for septate uterus in infertile patients before in vitro fertilization-embryo transfer (IVF-ET). The data of 937 infertile patients with septate uterus and achieved singleton pregnancy after IVF-ET from January 2014 to December 2015 were retrospectively analyzed. Thousand five hundred seventy-eight infertile patients with a normal uterus who achieved singleton pregnancy during the same period were selected as the control group. Patients with septate uterus were divided into two groups according to whether the septum was resected. The pregnancy and perinatal outcomes of the surgical group and the nonsurgical group were compared with the control group. The secondary infertility patients who were surgically corrected septa were also chosen as self-controls and an analysis was performed on their fertility outcomes pre- and post-surgery. Compared with the control group, the surgical group had increased rates of early miscarriage, preterm delivery, and low birthweight and a significantly reduced live birth rate (P < .05). The outcomes of the nonsurgical and control groups were similar. Using secondary infertility patients who were surgically corrected septa as self-controls, after surgery, the rates of miscarriage and ectopic pregnancy were significantly lower and the live birth rate was significantly higher (P ≤ .001); however, perinatal mortality was not significantly different before and after surgery. Patients with a septum depth greater than 10 mm or 5-10 mm associated with a history of unexplained recurrent miscarriage, IVF failure, or infertility might benefit from resection of the uterine septum with hysteroscopic metroplasty.


Correction to: Immunotherapy Alone or in Combination with Chemotherapy as First-Line Treatment of Non-Small Cell Lung Cancer.
The original version of this article unfortunately contained mistakes. In Table 2, under the column 'Lead to death' in Row 5 [CheckMate-026], the figures should read as '0.7' for Experimental Arm and '1.1' for Comparator. Right now, these are printed as 0.007 and 0.011 respectively.


Alternative pulse shapes in electrical hearing.
Cochlear implants (CIs) stimulate the auditory nerve with trains of symmetric biphasic (BI) pulses. We review studies showing that more efficient stimulation can be achieved by modifying these pulses by (1) increasing the inter-phase gap (IPG) between the two phases of each pulse, thereby delaying the recovery of charge, (2) increasing the duration and decreasing the amplitude of one phase - so-called "pseudomonophasic (PS)" waveforms, and (3) combining the pseudomonophasic stimulus with an IPG in a "delayed pseudomonophasic" waveform (PS_IPG). These efficiency gains, measured using changes in threshold and loudness, occur at a wide range of pulse rates, including those commonly used in current CI systems. In monopolar mode, dynamic ranges are larger for PS and for long-IPG pulse shapes than for BI pulses, but this increase in DR is not accompanied by a higher number of discriminable loudness steps, and hence, in a better coding of loudness. Moreover, waveforms with relatively short and long interphase gaps do not yield different patterns of excitation despite the relatively large differences in threshold. Two important findings are that, contrary to data obtained in animal experiments, anodic currents are more effective than cathodic stimulation for human CI patients and that the thresholds decrease with increases in IPG over a much longer time course (more than 3 ms) than for animals. In this review it is discussed how these alternative pulse shapes may be beneficial in terms of reducing power consumption and channel interactions, which issues remain to be addressed, and how models contribute to guiding our research.


Altered gene expression in human brain microvascular endothelial cells in response to the infection of influenza H1N1 virus.
Influenza viruses not only cause respiratory illness, but also have been reported to elicit neurological manifestations following acute viral infection. The central nervous system (CNS) has a specific defense mechanism against pathogens structured by cerebral microvasculature lined with brain endothelial cells to form the blood-brain barrier (BBB). To investigate the response of human brain microvascular endothelial cells (hBMECs) to the Influenza A virus (IAV), we inoculated the cells with the A/WSN/33 (H1N1) virus. We then conducted an RNAseq experiment to determine the changes in gene expression levels and the activated disease pathways following infection. The analysis revealed an effective activation of the innate immune defense by inducing the pattern recognition receptors (PRRs). Along with the production of proinflammatory cytokines, we detected an upregulation of interferons and interferon-stimulated genes, such as IFN-β/λ, ISG15, CXCL11, CXCL3 and IL-6, etc. Moreover, infected hBMECs exhibited a disruption in the cytoskeletal structure both on the transcriptomic and cytological levels. The RNAseq analysis showed different pathways and candidate genes associated with the neuroactive ligand-receptor interaction, neuroinflammation, and neurodegenerative diseases, together with a predicted activation of the neuroglia. Likewise, some genes linked with the mitochondrial structure and function displayed a significantly altered expression. En masse, this data supports that hBMECs could be infected by the IAV, which induces the innate and inflammatory immune response. The results suggest that the influenza virus infection could potentially induce a subsequent aggravation of neurological disorders. The online version contains supplementary material available at 10.1186/s44149-022-00053-9.


Pattern of nonspecific (or global) DNA methylation in oral carcinogenesis.
Although alterations in nonspecific (or global) DNA methylation (GDM) in specific cells are known to be involved in the process of lung carcinogenesis, similar associations have not been evaluated in other smoking-related cancers of the head and neck. We evaluated the status of GDM by using monoclonal antibodies specific for 5-methylcytosine (5-mc) in oral squamous cell carcinoma (SCC) specimens of 48 cigarette smokers who had SCC develop and in 93 age-, race-, and sex-matched smokers who did not. Percentages of cells positive for 5-mc immunostaining of DNA of SCC and dysplastic lesions were significantly higher than those of normal oral epithelial cells from cancer subjects and from noncancer subjects. The degree of DNA methylation was unrelated to DNA content. The pattern of GDM in oral SCCs is different from that of lung SCCs. The differences in nutrient risk factor profiles that are related to GDM and differential activity of DNA methyltranferases between oral and lung SCCs may explain these observations.


Intestinal alkaline phosphatase modulation by food components: predictive, preventive, and personalized strategies for novel treatment options in chronic kidney disease.
Alkaline phosphatase (AP) is a ubiquitous membrane-bound glycoprotein that catalyzes phosphate monoesters' hydrolysis from organic compounds, an essential process in cell signaling. Four AP isozymes have been described in humans, placental AP, germ cell AP, tissue nonspecific AP, and intestinal AP (IAP). IAP plays a crucial role in gut microbial homeostasis, nutrient uptake, and local and systemic inflammation, and its dysfunction is associated with persistent inflammatory disorders. AP is a strong predictor of mortality in the general population and patients with cardiovascular and chronic kidney disease (CKD). However, little is known about IAP modulation and its possible consequences in CKD, a disease characterized by gut microbiota imbalance and persistent low-grade inflammation. Mitigating inflammation and dysbiosis can prevent cardiovascular complications in patients with CKD, and monitoring factors such as IAP can be useful for predicting those complications. Here, we review IAP's role and the results of nutritional interventions targeting IAP in experimental models to prevent alterations in the gut microbiota, which could be a possible target of predictive, preventive, personalized medicine (PPPM) to avoid CKD complications. Microbiota and some nutrients may activate IAP, which seems to have a beneficial impact on health; however, data on CKD remains scarce.


Initial experience with hemostatic fibrin glue as adjuvant during drainless parotidectomy.
To evaluate the use of Hemostatic Fibrin Glue in parotidectomy without the use of surgical drains. Prospective cohort study of 10 patients undergoing parotidectomy. Surgery as a one day admission without the use of surgical drains was planned. The complication and duration of the hospital stay were obtained. Parotidectomy was undertaken by one surgeon. Prior to wound closure, the skin flap and wound bed were approximated using Tisseel tissue sealant. Data regarding the incidence of any complication and the duration of the hospital stay were obtained. Patients were followed to assess surgical outcome and document any complications. The mean follow-up period was 8 months (range 4-12 months). There were no major surgical complications. Two patients had facial nerve weakness due to adherence of the tumour in the facial nerve, in there was which complete recovery after few months. All patients were discharged the next day. None of the patients felt that the discharge had been premature. Parotidectomy can be undertaken safely without the need for surgical drains, therefore, allowing the patients to leave the hospital on the first postoperative day.


Albumin nanocomposites with MnO2/Gd2O3 motifs for precise MR imaging of acute myocardial infarction in rabbit models.
The severe mortality and morbidity of myocardial infarction requests appropriate and accurate detection. Considering pathological profile of the acidic myocardial infarction microenvironments, herein, the low pH-sensitive albumin nanocomposites with MnO2 motifs (MnO2@BSA) have been engineered for T1-weighted MR imaging of myocardial infarction, while using non-pH-responsive Gd2O3@BSA nanocomposites as control. The nanocomposites were 20-30 nm in diameter with spheroid morphology. Besides, the MnO2@BSA have exhibited pH-triggered releasing of Mn2+, demonstrating approximately 38-fold and 55-fold increased molecular relaxivity at acute myocardial infarction-mimicking pH 6.5 (13.08 mM-1s-1) and macrophage intracellular pH 5.0 (18.76 mM-1s-1) compared to the extremely low relaxivity (0.34 mM-1s-1) at normal physiological conditions (pH 7.4). However, the Gd2O3@BSA with molecular relaxivity approximately 10 mM-1s-1 were without pH-sensitive properties. Furthermore, the MnO2@BSA have demonstrated high accumulation in the acute myocardial infarction regions and fast metabolism from the body after systemic injection, accounting high contrast enhancement for accurate MR imaging of acute myocardial infarction in rabbit models, demonstrating better diagnostic performance over the controls.


Diagnosis of 21-hydroxylase deficiency in newborn infants by GC-MS of urinary steroids.
In a study using gas chromatography-mass spectrometry (GC-MS) on urine specimens from 16 normal infants and 16 infants with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (aged 1 day to 4 weeks), the major steroids recognized in all infants were: 16 alpha-hydroxy-dehydroepiandrosterone, 16 beta-hydroxy-dehydroepiandrosterone, 16-oxo-androstenediol, androstenetriol, 15 beta,17 alpha-dihydroxy-pregnenolone and 16 alpha-hydroxy-pregnenolone. Pregnanetriol was detectable in three normal infants (aged 3, 6 and 15 days) but the levels seen in 15 CAH patients were in a higher range. Pregnanetriolone, 5 beta-17-hydroxy-pregnanolone and 15 beta,17 alpha-dihydroxy-pregnanolone were present in the urine of 15 CAH patients, but were not detectable in any of the normal infants. The older the patient, the higher the level was of each of these four steroids. The results indicate that, even on day 1, patients with CAH due to 21-hydroxylase deficiency may be positively identified using GC-MS of urine specimens. This does not preclude the possibility that a minority of patients with CAH, most likely those with mild 21-hydroxylase deficiency, may not exhibit the characteristic GC-MS findings on day 1, as seen in one of the 16 CAH patients.


Short photoperiod accelerates preoptic and ventral telencephalic salmon GnRH synthesis and precocious maturation in underyearling male masu salmon.
The temporal relationship between testicular maturation and salmon gonadotropin-releasing hormone (sGnRH) mRNA expression was investigated in underyearling precocious male masu salmon, Oncorhynchus masou. Testicular maturation could be experimentally manipulated by changing the length of the light-dark photoperiod; maturation was accelerated in the short photoperiod group (8L-16D) and delayed in the long photoperiod group (16-8D). sGnRH mRNA and total silver grains in these loci in individual fish, increased with advancing testicular maturation. They were maximal in the short photoperiod group in August and in the long photoperiod group in September, when spermiation occurred. In contrast, marked changes in sGnRH synthetic activity in relation to testicular maturation were not observed in the terminal nerve ganglion or in the olfactory bulbs. sGnRH neurons in the preoptic area and the ventral telencephalon are clearly influenced by photoperiod and are involved in the control of gonadal maturation probably via gonadotropin secretion.


Review of recent clinical trials and their impact on the treatment of hypercholesterolemia.
Cardiovascular disease is the leading cause of death in the United States with incidence expected to increase in the coming decades. Recent years have produced a variety of new and novel therapeutics aimed at reducing the global burden of cardiovascular disease. This review highlights these recent advancements. In addition to more rigorous therapeutic thresholds for traditional LDL lowering agents such as statins, recent studies have developed new pathways of lipid lowering for both typical cardiovascular disease and complex, genetic lipid disorders. This includes inhibition of the cholesterol synthesis enzyme ATP citrate lyase with bempedoic acid, prevention of PCSK9 mRNA translation with inclisiran, inhibition of the lipoprotein lipase inhibitor angiopoetin like 3 protein with evinacumab and the use of anti-sense oligonucleotides to lower lipoprotein(a) levels. Icosapent ethyl, while remaining a topic of debate and controversy, demonstrates efficacy in cardiovascular risk reduction when all available data are examined. Lastly fibrate therapy continues to produce negative results in terms of cardiovascular disease reduction. Recent years have yielded breadth and depth to cardiovascular treatments. This expanded armamentarium will allow for more effective and more consistent treatment and prevention of cardiovascular disease.


Multiple signaling pathways are activated during insulin-like growth factor-I (IGF-I) stimulated breast cancer cell migration.
In order to display the full metastatic phenotype, the cancer cell must acquire the ability to migrate. In breast cancer, we have previously shown that insulin-like growth factor I (IGF-I) enhances cell motility in the highly metastatic MDA-231BO cell line by activating the type I IGF receptor (IGF1R). This motility response requires activation of IRS-2 and integrin ligation. In order to identify the key molecules downstream of IRS-2, we examined several signaling pathways known to be involved in cell motility. Focal adhesion kinase (FAK) was not activated by IGF-I, but IGF-I caused redistribution of FAK away from focal adhesion plaques. IGF-I treatment of MDA-231BO cells activated RhoA and inhibition of Rho-kinase (ROCK) inhibited the IGF-mediated motility response. The mitogen activated protein kinase (MAPK), p38, was also activated by IGF-I and inhibition of p38 by SB203580 blocked IGF-I induced cell motility. ROCK inhibition with Y-27632 also inhibited p38 phosphorylation suggesting that p38 lies downstream of ROCK. Both Erk1,2 and phosphatidyl-3 kinase (PI3K) were required for IGF-I stimulated cell motility, but only PI3K appeared to be directly downstream of IGF-I. Thus, IGF-I activation of its receptor coordinates multiple signaling pathways required for cell motility. Defining the key molecules downstream of the type I IGF receptor may provide a basis for optimizing therapies directed at this target.


Periodate or glutaraldehyde for preparing peroxidase conjugates?
Conjugates of horseradish peroxidase with antibodies (anti-human IgG (H + L)) or their Fab' fragments were prepared according to the newest modification of the periodate (P-) method or the two-step glutaraldehyde (G-) method. The conjugates were analysed by gel chromatography and subsequently tested in three different applications. For tissue immunohistochemistry on sections of lupus erythematosus skin, G-conjugates were preferred to polymeric P-conjugates. In ELISA for detection of human antibodies against penicillin P-conjugates were superior to G-conjugates. For the detection of surface Ig on lymphoid cells both types of conjugate were more or less equally suitable. A scheme for the most suitable combinations of method of preparation and field of application is given.


Management of severe congenital laryngeal webs - a 12 year review.
The aim of this study was to investigate respiratory and voice outcomes after open surgery for severe congenital laryngeal web. Included were all patients treated for severe congenital glottic web (Cohen type 3 and 4) between 2002 and 2014. Demographic data, symptoms, endoscopic findings, type of operation, outcome and complications were analyzed. Open surgery was performed with division of the web and enlargement of the subglottis by a laryngotracheal reconstruction or an extended partial cricotracheal resection. Fourteen patients were included. Information on the pre-operative voice was available in 11 patients. Aphonia was present in 6 patients (55%), dysphonia was severe in 1 (9%), moderate in 2 (18%) and mild in 2 (18%). Median age at operation was 19.3 months. As a first line treatment, laryngotracheal reconstruction was performed in 12 patients, and an extended partial cricotracheal resection in 2. Median follow-up was 9.5 months. All patients were successfully decannulated in a median time of 4 months after first surgery. Voice was improved in 10/11 (91%). Post-operatively, 3 patients (27%) had good voice. Dysphonia was severe in 2 (18%), moderate in 3 (27%) and mild in 3 patients (27%). Open surgery for Cohen type 3 and 4 glottic web allowed establishing a patent airway with successful decannulation in all cases. Quality of voice was improved in 91%, however, voice results were variable. In our opinion a delicate operative technique described in the paper is important for optimal voice results.


Explore the mechanism and substance basis of Mahuang FuziXixin Decoction for the treatment of lung cancer based on network pharmacology and molecular docking.
Mahuang FuziXixin Decoction (MFXD) is a classic Chinese herbal formula for the treatment of lung cancer. However, its mechanisms of action are unclear. In present study, network pharmacology and molecular docking technology were employed to investigate the molecular mechanism and substance basis of MFXD for the treatment of lung cancer. The active compounds and corresponding targets of MFXD were collected through the TCMSP database. OMIM and GeneCards databases were applied to filter the targets of lung cancer. The protein-protein interaction (PPI) were acquired through the STRING platform. Metascape and the Bioinformatics server were used for the visualization of GO and KEGG analysis. The tissue and organ distribution of targets was evaluated based on the BioGPS database. The binding affinity between potential targets and active compounds was evaluated by molecular docking. A total of 51 active compounds and 118 targets of MFXD were collected. The target with a higher degree were identified through the PPI network, namely AR, RELA, NCOA1, EGFR, FOS, CCND1, ESR1 and HSP90AA1. GO and KEGG analysis suggested that MFXD treatment of lung cancer mainly involves hormone and response to inorganic substance, transcription regular complex, transcription factor binding and Pathways in cancer. Experimental validation showed that MFXD treatment inhibited the proliferation of NSCLC cells through downregulation the expression of EGFR, HIF1A, NCOA1 and RELA. Moreover, molecular docking revealed that hydrogen bond and hydrophobic interaction contribute to the binding of the compounds to targets. Our findings comprehensively elucidated the actives, potential targets, and molecular mechanisms of MFXD against lung cancer, providing a promising strategy for the scientific basis and therapeutic mechanism of traditional Chinese medicine prescriptions for the treatment of the disease.


Concentration-controlled particle focusing in spiral elasto-inertial microfluidic devices.
Herein, we proposed a strategy for controlling the particle focusing position in Dean-coupled elasto-inertial flows via adjusting the polymer concentration of viscoelastic fluids. The physics behind the control strategy was then explored and discussed. At high polymer concentrations, the flowing particles could be single-line focused exactly at the channel centerline under the dominated elastic force. The center-line focusing in our spiral channel may employed as a potential pretreatment scheme for microflow cytometry detection. With further decreasing polymer concentrations, the particles would shift into the outer channel region under the comparable competition between inertial lift force, elastic force and Dean drag force. Finally, the observed position-shifting was successfully employed for particle concentration at a throughput much higher than most existing elasto-inertial microfluidics.


Chitosan-based nanomaterials: a state-of-the-art review.
This manuscript briefly reviews the extensive research as well as new developments on chitosan based nanomaterials for various applications. Chitosan is a biocompatible and biodegradable polymer having immense structural possibilities for chemical and mechanical modification to generate novel properties and functions in different fields especially in the biomedical field. Over the last era, research in functional biomaterials such as chitosan has led to the development of new drug delivery system and superior regenerative medicine, currently one of the most quickly growing fields in the area of health science. Chitosan is known as a biomaterial due to its biocompatibility, biodegradability, and non-toxic properties. These properties clearly point out that chitosan has greater potential for future development in different fields of science namely drug delivery, gene delivery, cell imaging, sensors and also in the treatment as well as diagnosis of some diseases like cancer. Chitosan based nanomaterials have superior physical and chemical properties such as high surface area, porosity, tensile strength, conductivity, photo-luminescent as well as increased mechanical properties as comparison to pure chitosan. This review highlights the recent research on different aspect of chitosan based nanomaterials, including their preparation and application.


Osteoporotic bone recovery by a bamboo-structured bioceramic with controlled release of hydroxyapatite nanoparticles.
While most bone defects can be repaired spontaneously, the healing process can be complicated due to insufficient bone regeneration when osteoporosis occurs. Synthetic materials that intrinsically stimulate bone formation without inclusion of exogenous cells or growth factors represent a highly desirable alternative to current grafting strategies for the management of osteoporotic defects. Herein, we developed a series of hydroxyapatite bioceramics composed of a microwhiskered scaffold (wHA) reinforced with multiple layers of releasable hydroxyapatite nanoparticles (nHA). These novel bioceramics (nwHA) are tunable to optimize the loading amount of nHA for osteoporotic bone formation. The utility of nwHA bioceramics for the proliferation or differentiation of osteoporotic osteoblasts in vitro is demonstrated. A much more compelling response is seen when bioceramics are implanted in critical-sized femur defects in osteoporotic rats, as nwHA bioceramics promote significantly higher bone regeneration and delay adjacent bone loss. Moreover, the nwHA bioceramics loaded with a moderate amount of nHA can induce new bone formation with a higher degree of ossification and homogenization. Two types of osteogenesis inside the nwHA bioceramic pores were discovered for the first time, depending on the direction of growth of the new bone. The current study recommends that these tailored hybrid micro/nanostructured bioceramics represent promising candidates for osteoporotic bone repair.


Nanostructuring Biomaterials with Specific Activities towards Digestive Enzymes for Controlled Gastrointestinal Absorption of Lipophilic Bioactive Molecules.
This review describes the development of novel lipid-based biomaterials that modulate fat digestion for the enhanced uptake of encapsulated lipophilic bioactive compounds (e.g. drugs and vitamins). Specific focus is directed towards analysing how key material characteristics affect the biological function of digestive lipases and manipulate lipolytic digestion. The mechanism of lipase action is a complex, interfacial process, whereby hydrolysis can be controlled by the ability for lipase to access and adsorb to the lipid-in-water interface. However, significant conjecture exists within the literature regarding parameters that influence the activities of digestive lipases. Important findings from recent investigations that strategically examined the interplay between the interfacial composition of the lipid microenvironment and lipolysis kinetics in simulated biophysical environments are presented. The correlation between lipolysis and the rate of solubilisation and absorption of lipophilic compounds in the gastrointestinal tract (GIT) is detailed. Greater insights into the mechanism of lipase action have provided a new approach for designing colloidal carriers that orally deliver poorly soluble compounds, directly impacting the pharmaceutical and food industries.


Effects of Cymbopogon citratus and Ferula assa-foetida extracts on glutamate-induced neurotoxicity.
Many of CNS diseases can lead to a great quantity of release of glutamate and the extreme glutamate induces neuronal cell damage and death. Here, we wanted to investigate the effects of Cymbopogon citratus essential oil and Ferula assa-foetida extracts treatment on glutamate-induced cell damage in a primary culture of rat cerebellar granule neurons. Cerebellums were collected from 7-d rat brains and cerebellar granule neurons were obtained after 8-d culture. CGN cells were treated with C. citratus essential oil and F. assa-foetida extracts at concentration of 100 μg/ml before, after, and during exposure to 30 μM glutamate. The cellular viability was evaluated by 3-(4, 5-dimethytthiazol-2-yl)-2, 5-diphenyltetrazoliumbromide (MTT) staining. The flow cytometry assay was used to examine cell cycle and apoptosis. MTT assay showed a glutamate-induced reduction in cellular viability while treatment with C. citratus essential oil and F. assa-foetida extracts before, during, and after exposure to glutamate was increased. Flow cytometric analysis indicated that F. assa-foetida extracts treatment significantly (p < 0.001) attenuated glutamate-induced apoptotic/necrotic cell death and the necrotic rate was decreased by C. citratus essential oil treatment compared to glutamate group, significantly (p < 0.001). The results show that C. citratus essential oil and F. assa-foetida extracts display neuroprotective effects in glutamate-induced neurotoxicity. These extracts exert antiapoptotic activity in cerebellar granule neurons due to cell cycle arrest in G0G1 phase, which explain the beneficial effects of C. citratus essential oil and F. assa-foetida extracts as therapies for neurologic disorders.


Protocol: a medium-throughput method for determination of cellulose content from single stem pieces of Arabidopsis thaliana.
Lignocellulosic biomass is an important renewable resource for biofuels and materials. How plants synthesise cellulose is not completely understood. It is known that cellulose synthase complex (CSCs) moving in the plasma membrane synthesise the cellulose. CESA proteins are the core components of CSC. In Arabidopsis, in vitro mutagenesis of proteins followed by complementation analysis of mutants lacking the gene represents an important tool for studying any biological process, including cellulose biosynthesis. Analysis of a large number of plants is crucial for these types of studies. By using aspiration rather than centrifugation to remove liquids during various stages of protocol, we were able to increase the throughput of the method as well as minimise the sample loss. As a test case, we determined cellulose content of wild type and secondary wall cesa mutants across the length of primary shoot which was fond to be rather uniform in 7-week-old plants. Additionally, we found that the cellulose content of single mutants was comparable to the higher order mutants. Here we describe a medium-throughput adaptation of Updegraff's method that allowed us to determine cellulose content of 200 samples each week.


High color rendering and high-luminance laser lighting using all inorganic nitride phosphor films.
Despite the huge advances that have been made in the development of ultra-high luminance laser lighting, achieving high color rendering properties in such systems at the same time remains a challenge. Recent studies show that in most cases, the luminous efficacy (LE) of laser lighting is compromised to improve the color rendering index (CRI). In this study, a possible solution to this problem has been proposed by preparing phosphor-in-glass (PiG) films comprised of the yellow-emitting phosphor (LSN:Ce3+) and the red-emitting phosphor (CASN:Eu2+). The composite material synthesized in this study exhibited outstanding optical and thermal properties. A uniform white light with a high CRI of 80.0 and a high LE of 185.9 lm W-1 was achieved by optimizing the yellow/red ratio and the emission peak position of the blue laser. Furthermore, it was found that this design enabled the phosphor to restrict the light emission area effectively, thus attaining a high luminous exitance of 1302 lm mm-2. With their superior optical performance, the PiG films can be regarded as promising color converter candidates for future high-quality laser-based white light sources.


Isolation of c-Kit+ human amniotic fluid stem cells from second trimester.
Amniotic fluid-derived stem (AFS) cells have been described as an appealing source of stem cells because of their (1) fetal, non-embryonic origin, (2) easy access during pregnancy overcoming the ethical issues related both to the use of human embryonic cells and to the postnatal tissue biopsy with donor site morbidity, and (3) their undemanding ability to be expanded. We and others have demonstrated the broad differentiation potential and here we describe the established protocol we developed to obtain c-Kit+ human AFS cells, starting from second trimester amniocentesis samples.


Bilateral renal cell carcinoma in a horseshoe kidney: preoperative assessment with MRI and digital subtraction angiography.
Renal cell carcinoma in a horseshoe kidney is an unusual entity. To our knowledge, only 123 cases have been published to date. We report the first bilateral case of two clear-cell carcinomas in an asymmetrically fused kidney. Optimum preservation of renal function after radical tumor removal requires accurate preoperative imaging. Since the vascular supply in fusion anomalies is extremely variable, angiography is mandatory. Magnetic resonance imaging was most suitable to predict the tumor extent and localization, because it simultaneously gave the most comprehensive anatomical overview of the malformation.


Familial lichen planus. A case report.
A Jewish Shirazee family is described in which 6 members had lichen planus. There was no increased incidence of the phenotype HLA A3.


Prevalence of carotid artery stenosis in patients with coronary artery disease in Japanese population.
Prevalence of carotid artery stenosis in patients with coronary artery disease (CAD) is unknown in Japanese population. The study populations consisted of 632 consecutive patients who underwent coronary angiography because of suspicion of CAD. All patients underwent carotid ultrasonography to screen carotid artery stenosis before coronary angiography. We defined echographic carotid stenosis as area stenosis of >50% or peak systolic velocity of >200 cm/s. Echographic carotid stenosis was observed in 124 patients (19.6%). Coronary angiography revealed 433 patients had CAD. Prevalence of echographic carotid artery stenosis was 14 of 199 (7.0%), 18 of 124 (14.5%), 28 of 131 (21.4%), and 64 of 178 (36.0%) in patients with 0-, 1-, 2-, and 3-vessel CAD, respectively (P<0.0001). The prevalence rate with carotid stenosis and CAD was 25.4%. Multivariate stepwise logistic regression analysis showed that age and the extent of CAD were independently related to the presence of carotid stenosis (P=0.0002 and <0.0001, respectively). Prevalence of carotid stenosis in patients with CAD is high in Japan as well as in Western countries. Screening of carotid artery stenosis is recommended especially in older patients with multivessel CAD.


A feasibility study using interactive graphic art feedback to augment acute neurorehabilitation therapy.
Interactive arts technologies, designed to augment the acute neurorehabilitation provided by expert therapists, may overcome existing barriers of access for patients with low motor and cognitive function. Develop an application prototype to present movement feedback interactively and creatively. Evaluate feasibility of use within acute neurorehabilitation. Record demographics and Functional Independent Measure™ scores among inpatients who used the technology during physical, occupational or recreational therapy. Record exercises performed with the technology, longest exercise duration performed (calculated from sensor data), user feedback, and therapist responses to a validated technology assessment questionnaire. Inpatients (n = 21) between the ages of 19 and 86 (mean 57 ± 18; 12 male/9 female) receiving treatment for motor deficits associated with neuropathology used the application in conjunction with occupational, recreational, or physical therapy during 1 to 7 sessions. Patients classified on the Functional Independence Measure™ as requiring 75%+ assistance for cognitive and motor function were able to use the interactive application. Customized interactive arts applications are appropriate for further study as a therapeutic modality. In addition to providing interactivity to individuals with low motor function, interactive arts applications might serve to augment activity-based medicine among inpatients with low problem-solving and memory function.


Large-Scale Complete Sequencing and Haplotyping of 1-10 kb DNA Molecules Using Short Massively Parallel Reads.
In this chapter, we describe a simple, low-cost method for making many copies of a single DNA molecule (1-10 kb in length) as a concatemer on a long DNA strand. This can enable applications requiring high-quality contiguous sequence and haplotype data from long single DNA molecules at large scale.


Increased striatal acetylcholine after 14 months cis-flupenthixol treatment in rats suggests functional supersensitivity of dopamine receptors.
Rats received continuous administration of cis-flupenthixol (0.8-1.2 mg/kg/day) or trans-flupenthixol (0.9-1.2 mg/kg/day) in drinking water for 14 months. The administration of cis-flupenthixol, but not trans-flupenthixol, caused apparent cerebral dopamine receptor supersensitivity. Thus, animals receiving cis-flupenthixol, but not trans-flupenthixol, showed enhanced apo-morphine-induced stereotyped behaviour. Dopamine concentration in striatum was not altered by drug treatment but striatal HVA and DOPAC concentrations were reduced in animals receiving cis-flupenthixol, but not trans-flupenthixol. No consistent change in Bmax of KD for specific striatal 3H-spiperone binding was observed after 14 months drug intake. However, in cis-flupenthixol treated animals a 40% increase in Bmax was observed following 2 weeks drug withdrawal. Continuous cis-flupenthixol intake increased striatal acetylcholine concentrations; trans-flupenthixol was without effect. This suggests the apparent increase in cerebral dopamine receptor supersensitivity caused by continuous long-term cis-flupenthixol administration is of functional importance in the intact animal.


Riociguat: Mode of Action and Clinical Development in Pulmonary Hypertension.
Pulmonary arterial hypertension (PAH) and chronic thromboembolic pulmonary hypertension (CTEPH) are progressive and debilitating diseases characterized by gradual obstruction of the pulmonary vasculature, leading to elevated pulmonary artery pressure (PAP) and increased pulmonary vascular resistance (PVR). If untreated, they can result in death due to right-sided heart failure. Riociguat is a novel soluble guanylate cyclase (sGC) stimulator that is approved for the treatment of PAH and CTEPH. We describe in detail the role of the nitric oxide-sGC-cyclic guanosine monophosphate (cGMP) signaling pathway in the pathogenesis of PAH and CTEPH and the mode of action of riociguat. We also review the preclinical data associated with the development of riociguat, along with the efficacy and safety data of riociguat from initial clinical trials and pivotal phase III randomized clinical trials in PAH and CTEPH.


Prostacyclin production in free versus native IMA grafts.
The long-term patency of the internal mammary artery (IMA) in coronary artery surgery has been superior to that of other grafts. We tested the IMAs as free grafts in an experimental study with 8 dogs. The right IMA was dissected free as a naked artery without surrounding tissue, and the left IMA was dissected with surrounding tissue using electrocautery. Both free IMA grafts were anastomosed first to the left IMA one after another and then to the left anterior descending coronary artery. The grafts were harvested 6 months later, and the prostacyclin (PGI2) production of the free IMA grafts and of the intact IMAs was measured. Radioimmunoassay was used to measure 6-keto-prostaglandin F1 alpha, the stable metabolite of PGI2. The PGI2 production of the right free IMA without surrounding tissue was 21.1 +/- 8.5 pg/mg and of the left IMA with surrounding tissue was 17.9 +/- 7.9 pg/mg (expressed in picograms of 6-keto-prostaglandin F1 alpha per milligram of tissue wet weight). The control value of the intact IMA was 18.3 +/- 6.5 pg/mg. Thus, PGI2 production of both free IMAs did not differ significantly from each other and from controls. This study showed that free IMAs were able to produce as much PGI2 as were control IMAs and that the type of dissection did not affect the result.


Susceptibility of avian influenza viruses of the N6 subtype to the neuraminidase inhibitor oseltamivir.
Avian influenza viruses are a source of genetic material that can be transmitted to humans through direct introduction or reassortment. Although there is a wealth of information concerning global monitoring for antiviral resistance among human viruses of the N1 and N2 neuraminidase (NA) subtypes, information concerning avian viruses of these and other NA subtypes is limited. We undertook a surveillance study to investigate the antiviral susceptibility of avian influenza N6 NA viruses, the predominant subtype among wild waterfowl. We evaluated 73 viruses from North American ducks and shorebirds for susceptibility to the NA inhibitor oseltamivir in a fluorescence-based NA enzyme inhibition assay. Most (90%) had mean IC(50) values ranging from <0.01 to 5.0nM; 10% were from 5.1 to 50.0nM; and none were >50.0nM. Susceptibility to oseltamivir remained stable among all isolates collected over approximately three decades (P⩽0.74). Two isolates with I222V NA substitution had moderately reduced susceptibility to oseltamivir in vitro (IC(50), 30.0 and 40.0nM). One field sample was a mixed population containing an avian paramyxovirus (APMV) and H4N6 influenza virus, as revealed by electron microscopy and hemagglutination inhibition assays with a panel of anti-APMV antisera. This highlights the importance of awareness and careful examination of non-influenza pathogens in field samples from avian sources. This study showed that oseltamivir-resistant N6 NA avian influenza viruses are rare, and must be tested both phenotypically and genotypically to confirm resistance.


The Combination of MR Elastography and Proton Density Fat Fraction Improves Diagnosis of Nonalcoholic Steatohepatitis.
Nonalcoholic fatty liver disease (NAFLD) is rapidly increasing worldwide. It is subdivided into nonalcoholic fatty liver (NAFL) and the more aggressive form, nonalcoholic steatohepatitis (NASH), which carries a higher risk of developing fibrosis and cirrhosis. There is currently no reliable non-invasive method for differentiating NASH from NAFL. To investigate the ability of magnetic resonance imaging (MRI)-based imaging biomarkers to diagnose NASH and moderate fibrosis as well as assess their repeatability. Prospective. Sixty-eight participants (41% women) with biopsy-proven NAFLD (53 NASH and 15 NAFL). Thirty participants underwent a second MRI in order to assess repeatability. 3.0 T; MR elastography (MRE) (a spin-echo echo-planar imaging [SE-EPI] sequence with motion-encoding gradients), MR proton density fat fraction (PDFF) and R2* mapping (a multi-echo three-dimensional gradient-echo sequence), T1 mapping (a single-point saturation-recovery technique), and diffusion-weighted imaging (SE-EPI sequence). Quantitative MRI measurements were obtained and assessed alone and in combination with biochemical markers (cytokeratin-18 [CK18] M30, alanine transaminase [ALT], and aspartate transaminase [AST]) using logistic regression models. Models that could differentiate between NASH and NAFL and between moderate to advanced fibrosis (F2-4) and no or mild fibrosis (F0-1), based on the histopathological results, were identified. Independent samples t-test, Pearson's chi-squared test, area under the receiver operating characteristic curve (AUROC), Spearman's correlation, intra-individual coefficient of variation, and intraclass correlation coefficient (ICC). Statistical significance was set at P < 0.05. There was a significant difference between the NASH and NAFL groups with liver stiffness assessed with MRE, CK18 M30, and ALT, with an AUROC of 0.74, 0.76, and 0.70, respectively. Both MRE and PDFF contributed significantly to a bivariate model for diagnosing NASH (AUROC = 0.84). MRE could significantly differentiate between F2-4 and F0-1 (AUROC = 0.74). A model combining MRE with AST improved the diagnosis of F2-4 (AUROC = 0.83). The ICC for repeatability was 0.94 and 0.99 for MRE and PDFF, respectively. MRE can potentially diagnose NASH and differentiate between fibrosis stages. Combining MRE with PDFF improves the diagnosis of NASH. 2 TECHNICAL EFFICACY: Stage 2.


Transcriptomic Profiling of Upper Tract Urothelial Carcinoma: Bladder Cancer Consensus Classification Relevance, Molecular Heterogeneity, and Differential Immune Signatures.
Analyses of large transcriptomics data sets of muscle-invasive bladder cancer (MIBC) have led to a consensus classification. Molecular subtypes of upper tract urothelial carcinomas (UTUCs) are less known. Our objective was to determine the relevance of the consensus classification in UTUCs by characterizing a novel cohort of surgically treated ≥pT1 tumors. Using immunohistochemistry (IHC), subtype markers GATA3-CK5/6-TUBB2B in multiplex, CK20, p16, Ki67, mismatch repair system proteins, and PD-L1 were evaluated. Heterogeneity was assessed morphologically and/or with subtype IHC. FGFR3 mutations were identified by pyrosequencing. We performed 3'RNA sequencing of each tumor, with multisampling in heterogeneous cases. Consensus classes, unsupervised groups, and microenvironment cell abundance were determined using gene expression. Most of the 66 patients were men (77.3%), with pT1 (n = 23, 34.8%) or pT2-4 stage UTUC (n = 43, 65.2%). FGFR3 mutations and mismatch repair-deficient status were identified in 40% and 4.7% of cases, respectively. Consensus subtypes robustly classified UTUCs and reflected intrinsic subgroups. All pT1 tumors were classified as luminal papillary (LumP). Combining our consensus classification results with those of previously published UTUC cohorts, LumP tumors represented 57.2% of ≥pT2 UTUCs, which was significantly higher than MIBCs. Ten patients (15.2%) harbored areas of distinct subtypes. Consensus classes were associated with FGFR3 mutations, stage, morphology, and IHC. The majority of LumP tumors were characterized by low immune infiltration and PD-L1 expression, in particular, if FGFR3 mutated. Our study shows that MIBC consensus classification robustly classified UTUCs and highlighted intratumoral molecular heterogeneity. The proportion of LumP was significantly higher in UTUCs than in MIBCs. Most LumP tumors showed low immune infiltration and PD-L1 expression and high proportion of FGFR3 mutations. These findings suggest differential response to novel therapies between patients with UTUC and those with MIBC.


Caudate clues to rewarding cues.
Behavioral studies indicate that prior experience can influence discrimination of subsequent stimuli. The mechanisms responsible for highlighting a particular aspect of the stimulus, such as motion or color, as most relevant and thus deserving further scrutiny, however, remain poorly understood. In the current issue of Neuron, demonstrate that neurons in the caudate nucleus of the basal ganglia signal which dimension of a visual cue, either color or location, is associated with reward in an eye movement task. These findings raise the possibility that this structure participates in the reward-based control of visual attention.


Choroid plexus cyst of the left lateral ventricle with intermittent blockage of the foramen of Monro, and initial invagination into the III ventricle in a child.
A cyst of the choroid plexus of the left lateral ventricle with intermittent blockage of the foramen of Monro and initially with invagination of the III ventricle in a child is described. In a 6-week-old boy a ventriculoatrial shunt was implanted for correction of an active asymmetrical hydrocephalus of unknown origin. When he was 3 months of age a water-soluble contrast CT ventriculography revealed a noncolloid cyst localised predominantly in the upper portion of the III ventricle. At that time the ventricular catheter obstructed with choroid plexus was removed; new bilateral catheters in a parieto-occipital region were implanted. In the course of the next 4 years, first the atrial catheter had to be extracted and then the peritoneal catheter was changed, in both cases because of obstruction. Periods of normal life alternated with periods of transient and intermittent symptoms of increased intracranial pressure, papilloedema, and myoclonic jerks. Repeated computed tomography (CT) and magnetic resonance imaging (MRI) showed stabilised hydrocephalus with an enlarged left lateral ventricle. When the boy was 16 years old MRI revealed a choroid plexus cyst in the left lateral ventricle 2 cm in diameter, with a ballvalve type of obstruction of the foramen of Monro. CT stereoendoscopic resection of the wall of a large cyst filled with cerebrospinal fluid was performed, and two additional adnexal small cysts were coagulated using the bipolar coagulator, Diomed 25 laser and scissors; the symptoms then regressed, except for superior bilateral altitudinal anopsia. Light and electron microscopy of the cyst wall is reported. The cyst was composed of collagenic connective tissue lined with a basal lamina lacking in epithelial cells. The preoperative and postoperative MRI are presented. Choroid plexus cysts localised in the anterior part of lateral ventricles are very rare, and all reported cases have been in male patients. According to the literature our case is only the third ever described in a child.


Modelling the nutritional ecology of ungulate herbivores: evolution of body size and competitive interactions.
A simulation model is used to quantify relationships between diet quality, digestive processes and body weight in ungulate herbivores. Retention time of food in the digestive tract is shown by regression to scale with W0.27, and to be longer in ruminants than in hindgut fermenters. Allometric relationships between whole gut mean retention time (MRT, h) and weight (W) were: MRT=9.4 W0.255 (r 2=0.80) for hindgut fermenters and MRT=15.3 W0.251 (r 2=0.76) in ruminants. Longer retention of ingesta by large-bodied ruminants and hindgut fermenters increases digestive efficiency relative to small animals and permits them to survive on lower-quality foods. Compared with ruminants, hindgut fermenters' faster throughput is an advantage which outweighs their lower digestive efficiency, particularly on poor quality foods, provided that food resources are not limiting. This suggests that the predominance of ruminants in the middle range of body weights results from their more efficient use of scarce resources under conditions of resource depletion. Considering only physical limitations on intake, the model shows that the allometric coefficient which scales energy intake to body mass is 0.88 in ruminants and 0.82 in hindgut fermenters. The advantages of large body size are countered by disadvantages where food quantity is limited, and we suggest that the upper limit to ungulate body size is determined by the ability to extract nutrients from feeding niches during the nadir of the seasonal cycle of resource quality and abundance.


EPR evidence that the M+ radical, which is observed in three site-directed mutants of photosystem II, is a tyrosine radical.
Isotopic labeling of Synechocystis sp. PCC 6803 and EPR spectroscopy have been used to demonstrate that photosystem II contains two redox active tyrosines, D and Z (Barry, B. A. and Babcock, G. T. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7099-7103; Boerner, R. J. and Barry, B. A. (1993) J. Biol. Chem. 268, 17151-17154). Another organic radical, M+, has recently been observed in site-directed mutants in which a non-redox active amino acid is substituted at either the putative D or putative Z sites (Boerner, R. J., Bixby, K. A., Nguyen, A. P., Noren, G. H., Debus, R. J., and Barry, B. A. (1993) J. Biol. Chem. 268, 1817-1823; Noren, G. H., and Barry, B. A. (1992) Biochemistry 31, 3335-3342). Here, we use isotopic labeling to determine the chemical identity of M+. Upon incorporation of perdeuterated tyrosine into photosystem II, the M+ EPR signal narrows to approximately 12-13 G. Labeling with 3,5-deuterated tyrosine results in an isotropic doublet with splittings of 11 G. Our results show that M+ is a tyrosine radical with unique spectroscopic properties.


[Optic neuropathy in positive anti-MOG antibody syndrome].
The diagnosis of optic neuritis (ON), or inflammation of the optic nerve, is based on clinical findings: first marked by rapidly progressive visual decline associated with eye pain accentuated by eye movements; abnormalities of color perception and/or contrast sensitivity may also be reported. In this case, inflammatory neuropathies are associated with anti-MOG antibodies. MOGs, oligodendrocytic glycoproteins involved in the production of myelin, were identified nearly three decades ago in association with demyelinating ON. The first series were reported in children following demyelinating neurological manifestations, particularly in ADEM (acute demyelinating encephalomyelitis) or multiple sclerosis (MS) [1]. Anti-MOGs are associated with neuropathies in the phenotypic setting of the neuromyelitis optica (NO) spectrum, and anti-Aquaporin 4 antibodies (AQP4) are negative by definition. Thus, anti-MOG could explain up to 30 % of cases of seronegative optic neuritis; their presence thus represents a significant diagnostic aid for the clinician, especially during a first neurological episode [1]. The first short published series in AQP4-/MOG+populations revealed primarily ophthalmological involvement with a good prognosis for recovery [1]. Knowledge of these antigens is important; it may permit not only an understanding of the physiopathology but also the stratification of patients in terms of prognosis and response to treatment [2]. Thus, the early diagnosis of anti-MOG positive ON must prompt aggressive initial treatment and a more or less maintenance therapy to prevent recurrence. The role of the ophthalmologist remains paramount, since most cases present with purely ocular involvement. We report herein the clinical, ophthalmological, laboratory and radiological data for 25 patients (45 eyes) managed between February 2011 and January 2017. All of our patients had optic neuritis associated with anti-MOG antibodies. All patients underwent the following testing: - Visual acuity; - Humphrey and/or Goldmann visual field; - Non-mydriatic fundus photography; - Optic disc OCT; - 3 Tesla orbital-cerebral MRI with and without contrast; - Standard and immunological laboratory testing for anti-MOG and anti AQP4 antibodies by Western Blot and ELISA. The male: female ratio of the population was 0.92 (13 women and 12 men). The average age at onset was 35.68 years (15 to 60 years); 40 % of the subjects were between 31 and 40 years old. The initial symptoms leading to consultation were mostly visual acuity (80 %) and pain (88 %). Involvement was bilateral in 80 % of cases (5 unilateral). Initial visual acuity was poor; 52 % of eyes were less than or equal to count fingers. The course was favorable however, with visual acuity returning to 10-12/10 after 6 months of follow-up (84 % of eyes). Orbital/cerebral MRI with attention to the visual pathways revealed involvement of the anterior visual pathways with gadolinium uptake in 92 % of cases. Of the 35 eyes initially considered affected, the main initial diagnoses were: - 36 % retro-bulbar optic neuritis (RBON); - 40 % anterior optic neuritis (AON); - 24 % other; of which 16 % were initially diagnosed as acute anterior ischemic optic neuropathy (AAION). 96 % of patients received corticosteroid treatment in the acute phase. 16 % required plasma exchange sessions. Maintenance therapy was proposed for only 36 % of the population. Optic neuritis is a pathology frequently encountered in ophthalmology; a good knowledge of symptoms and clinical signs is essential for early diagnosis and optimal management. The identification of autoantibodies, including anti-MOG antibodies, is important for patient management and is part of the required testing for all cases of optic neuritis, in order to adapt the treatment of the acute episode and to provide maintenance therapy to avoid recurrence.


Behavioral processes underlying the intake suppressive effects of melanocortin 3/4 receptor activation in the rat.
Central application of MTII, a melanocortin 3/4 receptor agonist, reduces food intake. The behavioral mechanisms underlying the anorexia, however, have not been evaluated. We examined the ingestive behavioral effects of MTII at the microstructural level using two complementary approaches. Rats were given daily 2-h sessions during which they drank 12.5% glucose solution; the time of occurrence of each lick event was recorded. We compared rats' glucose intake 30 min after the fourth ICV injection of 0.1, 0.33, and 1.0 nmol MTII or vehicle. The licking patterns were examined to discern effects on parameters related to taste processes and others related to post-ingestive inhibitory feedback. A second experiment directly analyzed the effect of MTII on motor performance by examining whether drug treated rats would, like controls, adjust licking output to maintain meal size when lick volume was shifted from 8 to 4 microl. Meal size was reduced by MTII in a dose-dependent manner (20-50%) in both experiments. Rats treated with MTII compensated for decreased lick volume by substantially increasing the number of licks emitted. Licking parameters associated with taste evaluation were not significantly affected by MTII, whereas parameters associated with post-ingestive inhibition varied as a function of treatment. Results suggest that MTII reduces intake by amplifying post-ingestive feedback inhibition. That MTII-treated rats increase the number of licks emitted in response to the lick volume reduction discounts the suggestion that intake inhibition is secondary to disruption of motor performance.


Comparative distribution of glyceraldehyde 3-phosphate dehydrogenase activity in human, guinea-pig, rabbit and mouse erythrocytes.
1. Guinea-pig erythrocytes contain half the activity of the enzyme glyceraldehyde-3-phosphate dehydrogenase (G3PD) present in human cells. About 60% of their total activity is membrane-bound. 2. Rabbit erythrocytes also contain half the amount of the enzyme of human red cells. The distribution of G3PD in rabbit cells, however, is similar to that of human cells with 70% of the enzyme being membrane-bound. 3. Mouse erythrocytes contain about two-thirds of G3PD activity present in human cells. All their enzyme activity is present in membrane-free hemolysate. 4. Non-human erythrocyte membrane proteins, in addition, have relatively greater amount of band 2.1, lack band 2.2, have a more heterogenous band 3 than its human counterpart, and have overlapping bands 4.1 and 4.2.


The medical, nutritional and surgical treatment of fistulae in Crohn's disease.
Of a total 44 patients with Crohn's disease, 10 patients with 9 internal and 15 external fistulae, some of which were recurrent, were analyzed at the Department of Surgery, Yokohama City University between 1973 and 1988. Twenty-two fistulae were treated with medical and nutritional therapy using either total parenteral or enteral hyperalimentation by which the closure rate of the internal and external fistulae was 0 (0/9) and 42 per cent (9/14), respectively. The nutritional status of all the patients with fistulae treated by nutritional therapy improved, especially those whose fistulae were closed. However, 8 of 9 internal fistulae and 5 of 15 external fistulae finally required resection of the fistula with the distal stenotic bowel segment. The re-opening rate of fistulae following successful medical/nutritional therapy and surgical therapy was 88.9 per cent (8/9) and 53.8 per cent (7/13), respectively, and the mean interval until recurrence was shorter in the patients who underwent medical and nutritional therapy (4.5 months) than in those who underwent surgical therapy (19.4 months). Thus, using medical and nutritional therapy, none of the internal fistulae were closed, but 9 of 14 external fistulae were. The optimal management of internal fistulae is therefore thought to be bowel resection to include the distal stenotic lesion, while medical and nutritional therapy is thought to be of value for external fistulae.


Carcinoid crisis after biopsy of a bronchial carcinoid.
Pulmonary carcinoids are rarely associated with carcinoid syndromes and even less commonly with carcinoid crisis. Somatostatin analogues can control carcinoid syndrome or crisis with tumors of gastrointestinal origin. We report the successful use of a somatostatin analogue in preventing carcinoid crisis at the time of resection of an "active" bronchial carcinoid tumor.