Miracle-dz/UniFAR-FTUs huggingface dataset:
A microtiter assay for activation of eosinophils

A simple microtiter assay for eosinophil activation is described. The assay used 1000–4000 eosinophils/microtiter well, and the design allows for a separate or simultaneous quantitative assessment of eosinophil adhension to protein‐coated microtiter wells and degranulation after stimulation with eosinophil‐activating factors. The number of adherent eosinophils is quantified indirectly by expressing the amount of eosinophil cationic protein (ECP) extracted from the adherent fraction of cells in percentage of the total amount of ECP extracted from the adherent fraction of cells in percentage of the total amount of ECP extracted from the cells added to the wells. Degranulation is quantified in the same way by expressing the amount of ECP or eosinophil protein X (EPX) released to the supernatant in percentage of the total amount of ECP or EPX. Known eosinophil‐activating agents such as PMA, interleukin (IL)‐3, IL‐5, GM‐CSF, and platelet‐activating factor (PAF) all induced a time‐ and does‐dependent adhension to albumin‐coated wells, whereas L_PAF did not. KInetic experiments showed that most adhesion occurred within the first 15–30 min, reaching a plateau around 60 min. After prolonged incubation, a decline in adhesion was detected. GM‐CSF‐induced adhesion was completely inhibited by incubation with monoclonal antibodies directed against the common β2‐chain (CD18) of the LFA‐1, Mac‐1, p150,95 complexes. Monoclonal antibodies against CD11a, CD11b, CD11c, VLA‐4 ALFA, ICAM‐1, VCAM‐1, Sialyl‐Lem, ELAM‐1, and LECAM had no inhibitory effect. Simultaneous monitoring of adhesion and degranulation after stimulation of eosinophils in albumincoated wells with either PMA or GM‐CSF showed that adhesion always preceded degranulation. Replacing the albumin coating of the microtiter wells with IgG or secretory IgA augmented both the spontaneous and the PMA‐ or GM‐CSF‐stimulated responses. In conclusion, the assay allows dynamic evaluation of eosinophil activationa dn can be used to assess soluble eosinophil‐activating factors as well as to study eosinophil activation by solidphase‐bound proteins.
Growth and wear of incisor and cheek teeth in domestic rabbits (Oryctolagus cuniculus) fed diets of different abrasiveness.

Although patterns of tooth wear are crucial in palaeo-reconstructions, and dental wear abnormalities are important in veterinary medicine, experimental investigations on the relationship between diet abrasiveness and tooth wear are rare. Here, we investigated the effect of four different pelleted diets of increasing abrasiveness (due to both internal [phytoliths] and external abrasives [sand]) or whole grass hay fed for 2 weeks each in random order to 16 rabbits (Oryctolagus cuniculus) on incisor and premolar growth and wear, and incisor and cheek tooth length. Wear and tooth length differed between diets, with significant effects of both internal and external abrasives. While diet abrasiveness was linked to tooth length for all tooth positions, whole forage had an additional effect on upper incisor length only. Tooth growth was strongly related to tooth wear and differed correspondingly between diets and tooth positions. At 1.4-3.2 mm/week, the growth of cheek teeth measured in this study was higher than previously reported for rabbits. Dental abnormalities were most distinct on the diet with sand. This study demonstrates that concepts of constant tooth growth in rabbits requiring consistent wear are inappropriate, and that diet form (whole vs. pelleted) does not necessarily affect cheek teeth. Irrespective of the strong effect of external abrasives, internal abrasives have the potential to induce wear and hence exert selective pressure in evolution. Detailed differences in wear effects between tooth positions allow inferences about the mastication process. Elucidating feedback mechanisms that link growth to tooth-specific wear represents a promising area of future research.
α-Catenin Is a Tumor Suppressor That Controls Cell Accumulation by Regulating the Localization and Activity of the Transcriptional Coactivator Yap1

A cell-cell adhesion protein suppresses skin carcinoma development by attenuating the signaling of a transcriptional coactivator in the Hippo pathway. Crowd Control for Skin Cells The adhesion protein α epithelial (αE) catenin has been implicated in limiting epithelial cell proliferation under conditions of high cell density (a process called contact inhibition of cell proliferation). Silvis et al. generated mice lacking αE-catenin in hair follicle stem cells and found that they developed squamous cell carcinomas, thus providing genetic evidence that αE-catenin is a tumor suppressor. The authors identified the transcriptional coactivator Yap1 as a protein that interacted with αE-catenin; this interaction inhibited the activity of Yap1 and prevented Yap1 from translocating to the nucleus. Yap1 is also a target for the Hippo signaling pathway, which restricts cell proliferation and organ size by blocking the nuclear translocation of Yap1. In human squamous cell carcinoma tumors, αE-catenin abundance was inversely correlated with Yap1 activation. Thus, αE-catenin may exert its tumor-suppressive effects by sequestering Yap1 and thereby limiting its activity. The Hippo pathway regulates contact inhibition of cell proliferation and, ultimately, organ size in diverse multicellular organisms. Inactivation of the Hippo pathway promotes nuclear localization of the transcriptional coactivator Yap1, a Hippo pathway effector, and can cause cancer. Here, we show that deletion of αE (α epithelial) catenin in the hair follicle stem cell compartment resulted in the development of skin squamous cell carcinoma in mice. Tumor formation was accelerated by simultaneous deletion of αE-catenin and the tumor suppressor–encoding gene p53. A small interfering RNA screen revealed a functional connection between αE-catenin and Yap1. By interacting with Yap1, αE-catenin promoted its cytoplasmic localization, and Yap1 showed constitutive nuclear localization in αE-catenin–null cells. We also found an inverse correlation between αE-catenin abundance and Yap1 activation in human squamous cell carcinoma tumors. These findings identify αE-catenin as a tumor suppressor that inhibits Yap1 activity and sequesters it in the cytoplasm.
Understanding quantitative pulsed CEST in the presence of MT

Phantom experiments in agar and ammonium chloride were performed to evaluate a three‐pool model of magnetization transfer and chemical exchange saturation transfer (CEST) in a pulsed saturation transfer experiment. The utility of the pulsed CEST method was demonstrated by varying the pH of the phantoms and observing the effect upon the CEST spectra both with and without the solid agar (the magnetization transfer pool), while fitting the spectra to the Bloch equation model with exchange. Pulsed CEST could be used to robustly quantify parameters related to CEST, including the exchange rate constant describing proton exchange with free water and the concentration of exchanging protons. Furthermore, the exchange rate constant and the CEST pool offset frequency of the ammonium chloride remained unchanged in the presence of a magnetization transfer pool. The logarithm of the fitted exchange rate constant was linearly related to pH: this relationship was maintained in the presence of magnetization transfer. Magn Reson Med, 2011. © 2011 Wiley‐Liss, Inc.
Exogenous Arachidonic Acid Promotes Insulin Release From Intact or Permeabilized Rat Islets by Dual Mechanisms: Putative Activation of Ca2+ Mobilization and Protein Kinase C

A number of indirect studies suggest a role for endogenous arachidonic acid (AA) in pancreatic islet function. To probe the effects of this fatty acid, AA and other polyunsaturated fatty acids were exogenously provided in Ca2+-free medium to avoid the formation of insoluble or impermeant Ca2+-arachidonate complexes. Concentrations of AA of ≧3 μM induced potent and sustained but reversible 45Ca efflux from prelabeled intact (or digitonin-permeabilized) islets; AA also nduced insulin release at somewhat higher concentrations. Other unsaturated fatty acids (erucic, oleic, linoleic, linolenic, dihomo-γ-linolenic, eicosapentaenoic, docosahexaenoic acids) were generally less active than AA itself, indicating a structure-function relationship. The effects of AA were saturable, were inhibitable by cooling, and were not accompanied in parallel by 51Cr release or trypan blue retention, suggesting a nontoxic mechanism. At low concentrations (3.3–16 μM), at which AA does not stimulate insulin release, AA-induced 45Ca efflux was not reduced by pretreatment with ionomycin (to deplete membrane-bound Ca2+ stores), suggesting stimulation of Ca2+ extrusion through the plasma membrane. At higher concentrations (≧25 μM), at which AA promotes insulin release, further Ca2+ efflux was stimulated, which was blunted by pretreatment with ionomycin (as well as by trifluoperazine). Conversely, pretreatment with AA obliterated the effects of ionomycin (3 μM) on cellular Ca2+ mobilization. Thus, AA also mobilizes Ca2+ from intracellular organelles, leading to a rise in free cytosolic Ca2+ (as previously reported). AA-induced 45Ca efflux and insulin release were independent of the presence of extracellular Na+ and did not require the oxygenation of AA. Dose-response curves comparing45Ca efflux and insulin secretion suggested that AA also stimulates hormone release by at least one other mechanism in addition to Ca2+ mobilization. This second stimulatory effect of AA could be seen in digitonin-permeabilized islets, where changes in cytosolic free Ca2+ concentration were vitiated by EGTA-containing buffers. Such secretion was also saturable and was inhibited by cooling or by spermine (which inhibits protein kinase C in the islet). Furthermore, AA-induced secretion from either intact or permeabilized islets was blunted by prolonged pretreatment of islets with a phorbol ester to deplete them of protein kinase C. Thus, exogenous arachidonic acid seems to be a complete secretagogue, having stimulatory effects both on Ca2+ mobilization and Ca2+-related secretory processes, putatively the activation of protein kinase C.
Effects of nonylphenol, bisphenol a, and their mixture on the viviparous swordtail fish (Xiphophorus helleri)

A number of fish species have been used for studies on endocrine disrupting chemicals (EDCs). However, despite the widespread use of oviparous fish, relatively little attention has been given to viviparous species. This study investigated the effects of EDCs in a viviparous fish and examined the possible usefulness of the fish as an alternative model for the studies on EDCs. Swordtails (Xiphophorus helleri) were exposed to nonylphenol (NP), bisphenol A (BPA), and their mixture. Both short‐term (3‐d) and relatively long‐term (60‐d) exposures were carried out using adult male and 30‐d‐old juvenile fish, respectively. Following the short‐term exposure, both NP and BPA caused vitellogenin mRNA expression. Flow cytometric analysis and terminal deox‐ynucleotidyl transferase assay on the testes of treated fish indicated reproductive damage. Histopathological analysis found degenerative and necrotic cells in seminiferous tubules following the exposure to 100 ppb NP. The testes with lesions were also associated with highly suppressed spermatogenesis. Following the long‐term exposure, both NP and BPA exposures significantly affected the growth of swordtails. In all cases, the results showed that the mixture was always more potent than a single chemical and that swordtail fish can be a useful model for the study of endocrine disruptors.
Percutaneous Endoscopic Gastrostomy for Long-Term Feeding of Patients With Oropharyngeal Tumors

Abstract: Secondary malnutrition in patients with oropharyngeal tumors can be the cause of substantial morbidity. The present, prospective study examined 30 patients receiving treatment for an oropharyngeal tumor. Percutaneous endoscopic gastrostomy (PEG) was performed 1 wk prior to the start of therapy in 15 patients, and 15 patients were temporarily fed by nasogastric tube after surgery. Anthropometric and laboratory parameters were measured in both patient groups at five defined times. Based on these parameters, the nutritional status of the PEG patients was found to be substantially better. The present study was able to show that the prophylactic placement of a PEG tube greatly improves the nutritional status of the patients before and after surgery and minimizes the catabolism of the body's fat depots.
On the detection of delay faults

The class of faults known as gate delay faults are investigated. A taxonomy of the classes of gate delay fault detecting tests is provided. Methods to derive robust and nonrobust tests to detect gate delay faults are proposed. A physically meaningful measure to assess the efficacy of test sequences is introduced, and used to report fault coverages. A nine-valued logic system was proposed and used for deriving these tests. A physically meaningful measure, in the form of the average detection size of a test sequence. An algorithm was devised and implemented to automate the process of test generation, and results of experimentation with the ATPG, as well as with a random-pattern simulator, on four ISCAS-85 circuits were reported. <<ETX>>
Characterization of mutations in the gene doublecortin in patients with double cortex syndrome

Mutations in the X‐linked gene doublecortin, which encodes a protein with no clear structural homologues, are found in pedigrees in which affected females show “double cortex” syndrome (DC; also known as subcortical band heterotopia or laminar heterotopia) and affected males show X‐linked lissencephaly. Mutations in doublecortin also cause sporadic DC in females. To determine the incidence of doublecortin mutations in DC, we investigated a cohort of eight pedigrees and 47 sporadic patients with DC for mutations in the doublecortin open reading frame as assessed by single‐stranded conformational polymorphism analysis. Mutations were identified in each of the eight DC pedigrees (100%), and in 18 of the 47 sporadic DC patients (38%). Identified mutations were of two types, protein truncation mutations and single amino acid substitution mutations. However, pedigrees with DC displayed almost exclusively single amino acid substitution mutations, suggesting that patients with these mutations may have less of a reproductive disadvantage versus those patients with protein truncation mutations. Single amino acid substitution mutations were tightly clustered in two regions of the open reading frame, suggesting that these two regions are critical for the function of the Doublecortin protein. Ann Neurol 1999;45:146–153
Treatments for hematologic malignancies in contrast to those for solid cancers are associated with reduced red cell alloimmunization

Red cell alloimmunization may induce severe hemolytic side effects. Identification of risk-modifying conditions will help tailor preventative strategies. This study aims to quantify the associations of hematologic malignancies and solid cancers with red cell alloimmunization in patients receiving red cell transfusions. We performed a nested multicenter case-control study in a source population of 24,063 patients receiving their first and subsequent red cell transfusions during an 8-year follow-up period. Cases (n=505), defined as patients developing a first transfusion-induced red cell alloantibody, were each compared with 2 non-alloimmunized controls (n=1010) who received a similar number of red cell units. Using multivariate logistic regression analyses, we evaluated the association of various malignancies and treatment regimens with alloimmunization during a delineated 5-week risk period. The incidence of alloimmunization among patients with acute (myeloid or lymphoid) leukemia and mature (B- or T-cell) lymphoma was significantly reduced compared to patients without these malignancies: adjusted relative risks (RR) with 95% confidence interval (CI) 0.36 (range 0.19–0.68) and 0.30 (range 0.12–0.81). Associations were primarily explained by immunosuppressive treatments [RR for (any type of) chemotherapy combined with immunotherapy 0.27 (95%CI: 0.09–0.83)]. Alloimmunization risks were similarly diminished in allogeneic or autologous stem cell transplanted patients (RR 0.34, 95%CI: 0.16–0.74), at least during the six months post transplant. Alloimmunization risks of patients with other hematologic diseases or solid cancers, and their associated treatment regimens were similar to risks in the general transfused population. Our findings suggest that, in contrast to malignancies in general, hemato-oncological patients treated with dose-intensive regimens have strongly diminished risk of red cell alloimmunization.
The bacterial signal molecule, ppGpp, regulates Salmonella virulence gene expression

Numerous, overlapping global regulatory systems mediate the environmental signalling controlling the virulence of Salmonella typhimurium. With both extra‐ and intracellular lifestyles, unravelling the mechanisms involved in regulating Salmonella pathogenesis has been complex. Here, we report a factor co‐ordinating environmental signals with global regulators involved in pathogenesis. An S. typhimuriumΔrelAΔspoT strain deficient in guanosine tetraphosphate (ppGpp) synthesis was found to be highly attenuated in vivo and non‐invasive in vitro. The ΔrelAΔspoT strain exhibited severely reduced expression of hilA and invF, encoding major transcriptional activators required for Salmonella pathogenicity island 1 (SPI‐1) gene expression and at least two other pathogenicity islands. None of the growth conditions intended to mimic the intestinal milieu was capable of inducing hilA expression in the absence of ppGpp. However, the expression of global regulators of Salmonella virulence, RpoS and PhoP/Q, and RpoS‐ and PhoP/Q‐dependent, non‐virulence‐related genes was not significantly different from the wild‐type strain. The results indicate that ppGpp plays a central role as a regulator of virulence gene expression in S. typhimurium and implicates ppGpp as a major factor in the environmental and host‐dependent regulation of Salmonella pathogenesis.
Further Studies on the Transmission of Staph. Aureus

of patients died from complications of endocarditis at about the time of completion of a six-weeks course of antibiotics adjudged to be curative, and, although no detailed study was made at necropsy concerning the viability of the bacterial colonies seen, their appearance gave no grounds for hope that, in these cases, several weeks less of treatment would have sufficed. In addition a case has already been quoted in which even six weeks was not long enough to effect cure. For penicillin-sensitive organisms fever that continues despite treatment may be due to systemic reaction to the antibiotic or to inadequate dosage. When considering this latter aspect it must be remembered that, while for streptococci of oral origin the bactericidal and bacteriostatic levels of penicillin are usually very similar, they are not always so, and this may explain failure of otherwise apparently adequate therapy (Berntsen, 1955). Cure of infection with so-called resistant organisms by the use of bacteriostatic drugs is not to be expected, and a drug or combination of drugs having bactericidal action must be sought (Garrod, 1953). The most generally useful combination at present is penicillin (10 to 20 mega units a day) with streptomycin 2 g. daily. But it is emphasized that treatment of such cases is part of a campaign planned in full co-operation with the bacteriologist, and rapid switching of drugs and dosage schedules merely serves to confuse the issue and may kill the patient. Should sensitivity reactions to the chosen bactericidal drug occur the lethal nature of bacterial endocarditis requires that rapid desensitization should be attempted in preference to reversion to treatment using bacteriostatic antibiotics.
RESPONSE OF BREAD WHEAT GENOTYPES TO DROUGHT SIMULATION UNDER A MOBILE RAIN SHELTER IN KENYA

Selection of drought tolerant wheat genotypes for Arid and Semi-arid lands (ASALS) of Kenya, which consist of 83% of total land area, can provide alternative agricultural land for expansion. To reduce cost of dryland research, simulated drought under a rain shelter offers a good alternative for screening because marginal areas are vast and widespread. Four moisture regimes which simulated terminal, early, mid- and late- season droughts were created under the mobile rain shelter at Njoro in 1998/99, by applying drip irrigation (i) up to seedling stage (70 mm) (ii) through tillering (82 mm) (iii) up to anthesis (94 mm) and (iv) grain filling (106 mm), respectively, to determine drought responses of five wheat varieties (Duma, R748, R830, R831 and R833) and identify drought tolerant genotypes. Control watering regime had 118 mm applied at all stages. Yield and yield components in each season and the two seasons combined showed significant difference (P Key Words: Duma, irrigation, dryland, moisture stress Resume La selection des genotypes tolerant a la secheresse pour les terres arides et semi-arides (ASALS) du Kenya, lesquelles consistent a 83% de la surface de terre, peut pourvenir de terre pour l'expansion agricole. Pour reduire le cout de la recherche dans les regions arides, la secheresse simulee a l'abris de la pluie offre une bonne alternative pour la selection parce que les aires marginales sont vastes et eparpillees. Pour les regimes humides qui sont simules a l'etat terminal, matinal, en pleine et en retard des saisons seches etaient crees sous un abri mobile a Njoro entre 1998/1999, par application de l'irrigation par goutte 1) jusqu'a l' etape de semis (70 mm) ii) a travers le labourage (82 mm), iii) jusqu'a l'etape d'anthere (94 mm) et iv) graine nourrissante (106 mm), respectivement pour determiner les reponses a la secheresse aux cinq varietes de ble (Duma, R748, R830, R831 et R833) et identifier les genotypes tolerant la secheresse. Le controle du regime en eau avait 118 mm appliques a toutes les etapes. Les composants de la production et du rendement dans chaque saison et les deux saisons combinees ont montre une difference significative (P Mots Cles: Duma, irrigation, terre seche, stress d'humidite (Af Crop Sci J 2003 Vol 11 No 3 pp225-234