SKOPE-IT (Shareable Knowledge Objects as Portable Intelligent Tutors): overlaying natural language tutoring on an adaptive learning system for mathematics.
This study investigated learning outcomes and user perceptions from interactions with a hybrid intelligent tutoring system created by combining the AutoTutor conversational tutoring system with the Assessment and Learning in Knowledge Spaces (ALEKS) adaptive learning system for mathematics. This hybrid intelligent tutoring system (ITS) uses a service-oriented architecture to combine these two web-based systems. Self-explanation tutoring dialogs were used to talk students through step-by-step worked examples to algebra problems. These worked examples presented an isomorphic problem to the preceding algebra problem that the student could not solve in the adaptive learning system. Due to crossover issues between conditions, experimental versus control condition assignment did not show significant differences in learning gains. However, strong dose-dependent learning gains were observed that could not be otherwise explained by either initial mastery or time-on-task. User perceptions of the dialog-based tutoring were mixed, and survey results indicate that this may be due to the pacing of dialog-based tutoring using voice, students judging the agents based on their own performance (i.e., the quality of their answers to agent questions), and the students' expectations about mathematics pedagogy (i.e., expecting to solving problems rather than talking about concepts). Across all users, learning was most strongly influenced by time spent studying, which correlated with students' self-reported tendencies toward effort avoidance, effective study habits, and beliefs about their ability to improve in mathematics with effort. Integrating multiple adaptive tutoring systems with complementary strengths shows some potential to improve learning. However, managing learner expectations during transitions between systems remains an open research area. Finally, while personalized adaptation can improve learning efficiency, effort and time-on-task for learning remains a dominant factor that must be considered by interventions.


Prophylaxis of affective disorders. Current status of knowledge.
A review of all properly controlled studies clearly indicates that lithium carbonate is prophylactic for mania in bipolar patients; it is suggestive of prophylaxis for depression in both bipolar and unipolar patients. Studies are outlined that would clarify lithium carbonate's prophylactic effect for depression in these two patient groups. Continuation therapy with antidepressants reduces incidences of recurrence in unipolar depressives. The only controlled study indicates that tricyclic antidepressants may have prophylactic effect in unipolar patients. This finding needs confirmation. Data are insufficient for conclusions on prophylactic treatment for schizoaffective disorders.


THE INCIDENCE OF NEOVASCULARIZATION IN CENTRAL SEROUS CHORIORETINOPATHY BY OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY.
To evaluate the incidence of neovascularization (NV) secondary to central serous chorioretinopathy (CSC)-a condition belonging to the spectrum of pachychoroid disorders by means of optical coherence tomography angiography. One hundred and seventy five eyes with CSC were evaluated in this retrospective observational study. The eyes with acute or chronic CSC with no NV were included in Group 1, and those with NV were evaluated in Group 2. Only eyes that had undergone structural optical coherence tomography and optical coherence tomography angiography were included. Age, best-corrected visual acuity, and subfoveal choroidal thickness were evaluated in all eyes. In Group 2, the type and morphology of NV and the occurrence of exudation were considered. Of a total of 175 eyes with CSC, 86 had the acute form and 89 the chronic. Approximately 140 belonged to Group 1 (80%) and 35 to Group 2 (20%). Approximately 39.2% of all patient with chronic CSC developed NV. Mean age in Groups 1 and 2 was 53.3 years (±10.9) and 66.6 years (±10.2), respectively. Mean best-corrected visual acuity in Groups 1 and 2 was 45.7 (±11.7) and 30.9 (±17.9) early treatment diabetic retinopathy study letters, respectively. Mean CCT in Group 1 and 2 was 417.5 µm (±123) and 344.2 µm (±165.9), respectively. In Group 2, all patients had Type 1 NV (100%); 29 eyes (83%) had filamentous feature, and 6 eyes (17%) had irregular shape. Silent nonexudative NV was observed in 7 eyes (20%), all belonging to Group 2. The use of optical coherence tomography angiography in everyday clinical practice allows for the accurate analysis of the chorioretinal vascular setting, with the identification of new vessels that could remain misdiagnosed.


Partitioning nuclear and chloroplast variation at multiple spatial scales in the neotropical epiphytic orchid, Laelia rubescens.
Insights into processes that lead to the distribution of genetic variation within plant species require recognition of the importance of both pollen and seed movement. Here we investigate the contributions of pollen and seed movement to overall gene flow in the Central American epiphytic orchid, Laelia rubescens. Genetic diversity and structure were examined at multiple spatial scales in the tropical dry forest of Costa Rica using nuclear (allozymes) and chloroplast restriction fragment length polymorphism (RFLP) markers, which were found to be diverse (allozymes, P = 73.3%; HE = 0.174; cpDNA, HE = 0.741). Nuclear genetic structure (FSTn) was low at every spatial scale (0.005-0.091). Chloroplast markers displayed more structure (0.073-0.254) but relatively similar patterns. Neither genome displayed significant isolation-by-distance. Pollen and seed dispersal rates did not differ significantly from one another (mp/ms = 1.40) at the broadest geographical scale, among sites throughout Costa Rica. However, relative contributions of pollen and seeds to gene flow were scale-dependent, with different mechanisms determining the dominant mode of gene flow at different spatial scales. Much seed dispersal is highly localized within the maternal population, while some seeds enter the air column and are dispersed over considerable distances. At the intermediate scale (10s to 100s of metres) pollinators are responsible for substantial pollen flow. This species appears capable of distributing its genes across the anthropogenically altered landscape that now characterizes its Costa Rican dry forest habitat.


Progress on diagnostic and prognostic markers of pancreatic cancer.
Pancreatic cancer is a malignant disease characterized by low survival and high recurrence rate, whose patients are mostly at the stage of locally advanced or metastatic disease when first diagnosed. Early diagnosis is particularly important because prognostic/predictive markers help guide optimal individualized treatment regimens. So far, CA19-9 is the only biomarker for pancreatic cancer approved by the FDA, but its effectiveness is limited by low sensitivity and specificity. With recent advances in genomics, proteomics, metabolomics, and other analytical and sequencing technologies, the rapid acquisition and screening of biomarkers is now possible. Liquid biopsy also occupies a significant place due to its unique advantages. In this review, we systematically describe and evaluate the available biomarkers that have the greatest potential as vital tools in diagnosing and treating pancreatic cancer.


Optimising adjacent membrane segmentation and parameterisation in multicellular aggregates by piecewise active contours.
In fluorescence microscopy imaging, the segmentation of adjacent cell membranes within cell aggregates, multicellular samples, tissue, organs, or whole organisms remains a challenging task. The lipid bilayer is a very thin membrane when compared to the wavelength of photons in the visual spectra. Fluorescent molecules or proteins used for labelling membranes provide a limited signal intensity, and light scattering in combination with sample dynamics during in vivo imaging lead to poor or ambivalent signal patterns that hinder precise localisation of the membrane sheets. In the proximity of cells, membranes approach and distance each other. Here, the presence of membrane protrusions such as blebs; filopodia and lamellipodia; microvilli; or membrane vesicle trafficking, lead to a plurality of signal patterns, and the accurate localisation of two adjacent membranes becomes difficult. Several computational methods for membrane segmentation have been introduced. However, few of them specifically consider the accurate detection of adjacent membranes. In this article we present ALPACA (ALgorithm for Piecewise Adjacent Contour Adjustment), a novel method based on 2D piecewise parametric active contours that allows: (i) a definition of proximity for adjacent contours, (ii) a precise detection of adjacent, nonadjacent, and overlapping contour sections, (iii) the definition of a polyline for an optimised shared contour within adjacent sections and (iv) a solution for connecting adjacent and nonadjacent sections under the constraint of preserving the inherent cell morphology. We show that ALPACA leads to a precise quantification of adjacent and nonadjacent membrane zones in regular hexagons and live image sequences of cells of the parapineal organ during zebrafish embryo development. The algorithm detects and corrects adjacent, nonadjacent, and overlapping contour sections within a selected adjacency distance d, calculates shared contour sections for neighbouring cells with minimum alterations of the contour characteristics, and presents piecewise active contour solutions, preserving the contour shape and the overall cell morphology. ALPACA quantifies adjacent contours and can improve the meshing of 3D surfaces, the determination of forces, or tracking of contours in combination with previously published algorithms. We discuss pitfalls, strengths, and limits of our approach, and present a guideline to take the best decision for varying experimental conditions for in vivo microscopy.


On the use of hyperpolarized helium MRI for conformal avoidance lung radiotherapy.
We wanted to illustrate the feasibility of using hyperpolarized helium magnetic resonance imaging (HPH-MRI) to obtain functional information that may assist in improving conformal avoidance of ventilating lung tissue during thoracic radiotherapy. HPH-MRI images were obtained from a volunteer patient and were first fused with a proton density-weighted (PD(w)) MRI to provide corresponding anatomic detail; they were then fused with the treatment planning computed tomography scan of a patient from our treatment planning database who possessed equivalent thoracic dimensions. An optimized treatment plan was then generated using the TomoTherapy treatment planning system, designating the HPH-enhancing regions as ventilation volume (VV). A dose-volume histogram compares the dosimetry of the lungs as a paired organ, the VV, and the lungs minus the VV. The clinical consequences of these changes was estimated using a bio-effect model, the parallel architecture model, or the local damage (f(dam)) model. Model parameters were chosen from published studies linking the incidence of grade 3+ pneumonitis, with the dose and volume irradiated. For two hypothetical treatment plans of 60 Gy in 30 fractions delivered to a right upper-lobe lung mass, one using and one ignoring the VV as an avoidance structure, the mean normalized total dose (NTD(mean)) values for the lung subvolumes were: lungs = 12.5 Gy₃ vs. 13.52 Gy₃, VV = 9.94 Gy₃ vs. 13.95 Gy₃, and lungs minus VV = 16.69 Gy₃ vs. 19.16 Gy₃. Using the f(dam) values generated from these plans, one would predict a reduction of the incidence of grade 3+ radiation pneumonitis from 12%-4% when compared with a conventionally optimized plan. The use of HPH-MRI to identify ventilated lung subvolumes is feasible and has the potential to be incorporated into conformal avoidance treatment planning paradigms. A prospective clinical study evaluating this imaging technique is being developed.


The small GTPase Cdc42 modulates the number of exocytosis-competent dense-core vesicles in PC12 cells.
Although the small GTPase Rho family Cdc42 has been shown to facilitate exocytosis through increasing the amount of hormones released, the precise mechanisms regulating the quantity of hormones released on exocytosis are not well understood. Here we show by live cell imaging analysis under TIRF microscope and immunocytochemical analysis under confocal microscope that Cdc42 modulated the number of fusion events and the number of dense-core vesicles produced in the cells. Overexpression of a wild-type or constitutively-active form of Cdc42 strongly facilitated high-KCl-induced exocytosis from the newly recruited plasma membrane vesicles in PC12 cells. By contrast, a dominant-negative form of Cdc42 inhibited exocytosis from both the newly recruited and previously docked plasma membrane vesicles. The number of intracellular dense-core vesicles was increased by the overexpression of both a wild-type and constitutively-active form of Cdc42. Consistently, activation of Cdc42 by overexpression of Tuba, a Golgi-associated guanine nucleotide exchange factor for Cdc42 increased the number of intracellular dense-core vesicles, whereas inhibition of Cdc42 by overexpression of the Cdc42/Rac interactive binding domain of neuronal Wiskott-Aldrich syndrome protein decreased the number of them. These findings suggest that Cdc42 facilitates exocytosis by modulating both the number of exocytosis-competent dense-core vesicles and the production of dense-core vesicles in PC12 cells.


Pharmacokinetics and first-pass metabolism of astaxanthin in rats.
Astaxanthin is a carotenoid with antioxidant, anti-cancer and anti-inflammatory properties. The pharmacokinetics of astaxanthin after its intravenous (5, 10, and 20 mg/kg) and oral (100 and 200 mg/kg) administration and its first-pass extraction ratios after its intravenous, intraportal or intragastric (20 mg/kg) administration were evaluated in rats. The pharmacokinetic parameters of astaxanthin were dose dependent after its intravenous administration, due to the saturable hepatic metabolism of astaxanthin, but dose independent after oral administration. The gastrointestinal absorption of astaxanthin followed the flip-flop model. The hepatic and gastrointestinal first-pass extraction ratios of astaxanthin were approximately 0·490 and 0·901, respectively. Astaxanthin was metabolised primarily by hepatic cytochrome P-450 1A1/2 in rats. Astaxanthin was unstable up to 4 h incubation in four rat gastric juices and up to 24 h incubation in various buffer solutions having a pH of 1-13. The tissue/plasma ratios of astaxanthin at 8 and 24 h after its oral administration (100 mg/kg) were greater than unity for all tissues studied, except in the heart, at 8 h, indicating that the rat tissues studied had high affinity for astaxanthin.


CSAR: a contig scaffolding tool using algebraic rearrangements.
Advances in next generation sequencing have generated massive amounts of short reads. However, assembling genome sequences from short reads still remains a challenging task. Due to errors in reads and large repeats in the genome, many of current assembly tools usually produce just collections of contigs whose relative positions and orientations along the genome being sequenced are still unknown. To address this issue, a scaffolding process to order and orient the contigs of a draft genome is needed for completing the genome sequence. In this work, we propose a new scaffolding tool called CSAR that can efficiently and more accurately order and orient the contigs of a given draft genome based on a reference genome of a related organism. In particular, the reference genome required by CSAR is not necessary to be complete in sequence. Our experimental results on real datasets have shown that CSAR outperforms other similar tools such as Projector2, OSLay and Mauve Aligner in terms of average sensitivity, precision, F-score, genome coverage, NGA50 and running time. The program of CSAR can be downloaded from https://github.com/ablab-nthu/CSAR. firstname.lastname@example.com or firstname.lastname@example.com. Supplementary data are available at Bioinformatics online.


Context and meter enhance long-range planning in music performance.
Neural responses demonstrate evidence of resonance, or oscillation, during the production of periodic auditory events. Music contains periodic auditory events that give rise to a sense of beat, which in turn generates a sense of meter on the basis of multiple periodicities. Metrical hierarchies may aid memory for music by facilitating similarity-based associations among sequence events at different periodic distances that unfold in longer contexts. A fundamental question is how metrical associations arising from a musical context influence memory during music performance. Longer contexts may facilitate metrical associations at higher hierarchical levels more than shorter contexts, a prediction of the range model, a formal model of planning processes in music performance (Palmer and Pfordresher, 2003; Pfordresher et al., 2007). Serial ordering errors, in which intended sequence events are produced in incorrect sequence positions, were measured as skilled pianists performed musical pieces that contained excerpts embedded in long or short musical contexts. Pitch errors arose from metrically similar positions and further sequential distances more often when the excerpt was embedded in long contexts compared to short contexts. Musicians' keystroke intensities and error rates also revealed influences of metrical hierarchies, which differed for performances in long and short contexts. The range model accounted for contextual effects and provided better fits to empirical findings when metrical associations between sequence events were included. Longer sequence contexts may facilitate planning during sequence production by increasing conceptual similarity between hierarchically associated events. These findings are consistent with the notion that neural oscillations at multiple periodicities may strengthen metrical associations across sequence events during planning.


Potentiation of ICI182,780 (Fulvestrant)-induced estrogen receptor-alpha degradation by the estrogen receptor-related receptor-alpha inverse agonist XCT790.
ICI182,780 (Fulvestrant) is a pure anti-estrogen used in adjuvant therapies of breast cancer. This compound not only inhibits the transcriptional activities of the estrogen receptor-alpha (ER alpha) but also induces its proteasome-dependent degradation. The latter activity is believed to be required for the antiproliferative effects of ICI182,780. Estrogen receptor-related receptor-alpha (ERR alpha) is an orphan member of the nuclear receptor superfamily that is expressed in a wide range of tissues including breast tumors, in which its high expression correlates with poor prognosis. Although not regulated by any natural ligand, ERR alpha can be deactivated by the synthetic molecule XCT790. Here we demonstrate that this compound also induces a proteasome degradation of ERR alpha. We also show that although it does not act directly on the steady-state level of ER alpha, XCT790 potentiates the ICI182,780-induced ER alpha degradation. We suggest that treatment with XCT790 could thus enhance the efficacy of ICI182,780 in ER alpha-dependent pathologies such as breast cancer.


The tiny Hairless protein from Apis mellifera: a potent antagonist of Notch signaling in Drosophila melanogaster.
The Notch signaling pathway is fundamental to the regulation of many cell fate decisions in eumetazoans. Not surprisingly, members of this pathway are highly conserved even between vertebrates and invertebrates. There is one notable exception, Hairless, which acts as a general Notch antagonist in Drosophila. Hairless silences Notch target genes by assembling a repressor complex together with Suppressor of Hairless [Su(H)] and the co-repressors Groucho (Gro) and C-terminal binding protein (CtBP). Now with the availability of genomic databases, presumptive Hairless homologues are predicted, however only in insect species. To further our understanding of Hairless structure and function, we have cloned the Hairless gene from Apis mellifera (A.m.H) and characterized its functional conservation in Drosophila. The Apis Hairless protein is only one third of the size of the Drosophila orthologue. Interestingly, the defined Suppressor of Hairless binding domain is interrupted by a nonconserved spacer sequence and the N-terminal motif is sufficient for binding. In contrast to Apis Hairless, the Drosophila orthologue contains a large acidic domain and we provide experimental evidence that this acidic domain is necessary to silence Hairless activity in vivo. Despite the dramatic size differences, Apis Hairless binds to the Drosophila Hairless interactors Su(H), Gro, CtBP and Pros26.4. Hence, Apis Hairless assembles a repressor complex with Drosophila components that may have a different topology. Nevertheless, Apis Hairless is sufficient to repress the Notch target gene vestigial in Drosophila. Moreover, it is able to rescue Hairless mutant phenotypes, providing in vivo evidence for its function as a bona fide Notch antagonist. This is the first interspecies-complementation analysis of the Hairless gene. Guided by evolutionary comparisons, we hope to eventually identify all the relevant structural domains and cofactors of Hairless, thereby opening an avenue for further insights into the repressor-complexes that down-regulate Notch signaling also in other, higher eukaryotes.


[Treatment of non-variceal hemorrhage of the upper digestive tract].
High digestive tract hemorrhage (HDTH) represents on average 35% of the indications for endoscopy. It shows as a complication in different digestive pathologies or secondary to coagulopathies. Endoscopic management of non-variceal HDTH includes a gamut of procedures that when grouped together, have shown to be effective and safe in its control with an important diminishing in morbidity-mortality, transfusion requirements, days of hospital stay, and the need for surgery. The most frequently employed methods are substance injections, multipolar coagulation, and thermic catheter, which achieve an average 90% hemostasis, but still inform high percentages of relapse. Approximately 25% of the patients with non-variceal hemorrhage may bleed again after endoscopic management. New procedures such as hemoclips and the combination of endoscopic methods have been used to diminish relapse. Endoscopic treatment should be carried out when we find bleeding lesions with Forrest Ia, Ib and IIa classifications, fundamentally because the risk of hemorrhagic relapse is very high among these groups. The treatment for the eradication of Helicobacter pylori in patients with bleeding peptic ulcers diminishes, in an important manner, the risk of the recurrence of hemorrhage.


Mini-Invasive Intramedullary Fixation of Displaced Midshaft Clavicle Fractures with an Elastic Titanium Nail.
The treatment of completely displaced midshaft clavicle fractures is still controversial, but surgical treatment provides a shorter recovery period and higher union rates than nonoperative treatment with a sling1-5. Even though the literature does not clearly support surgery for these fractures, surgery is becoming more frequent6. Elastic stable intramedullary nailing (ESIN) with a titanium nail is a well-documented mini-invasive procedure with functional outcomes comparable with those of plate fixation after 1 year but a high rate of implant removal of up to 80%6-8. The ideal injury for ESIN is a fracture without comminution that is available for surgery within a couple of days. ESIN is also a good option for fractures with comminution, but a somewhat slower functional recovery during the first 6 months should be expected. The patient is placed in beach-chair position with the fluoroscope placed on the injured side or cranially in relation to the fracture. The surgeon pushes the skin gently with his/her index finger at the jugular fossa before incising it. This places the skin incision inferior to the implant. A 1 to 2-cm incision down to the bone is then placed about 1 to 2 cm lateral to the sternoclavicular joint. A unicortical entry hole is made at the medial end with a 2.5-mm drill bit and then widened and directed laterally with an awl. Two reduction forceps are placed percutaneously and used by the assistant to align the main fragments. The nail is passed with oscillating movements until it is secured into the lateral fragment. If it is not possible to obtain a closed reduction, an open reduction is performed. The nail is cut short down to the bone, and the myocutaneous layer and skin are closed with sutures. Five to 10 mL of bupivacaine is injected into the fracture area for postoperative pain management. The arm is placed in a simple sling for comfort. A non-weight-bearing active range of motion of <90° is encouraged. Closed completely displaced midshaft clavicle fractures can be treated operatively or nonoperatively with a sling. Superior plate fixation is well documented, but use of anterior or several mini-fragment plates is also an option. There are several intramedullary implant techniques, but most require an open reduction. Most intramedullary implant methods are open procedures, whereas closed reduction and internal fixation is possible with ESIN. This ensures preservation of the fracture hematoma and no periosteal stripping, in contrast with the open intramedullary techniques. The nail enters the fracture medially, which decreases the chance of perioperative pulmonary or neurovascular injury. The procedure is less time-consuming than plate fixation, the result is cosmetically superior, and the functional outcomes after 1 year are equal. When ESIN is used for fractures without intermediary fragments, the functional recovery time is equal to that after plate fixation, although it is longer when used for fractures with intermediary fragments8.


[siRNA-mediated silencing of ClC-2 gene inhibits proliferation of human U-87 glioma cells].
Small interfering RNA (siRNA), which has been used to inhibit mammalian gene expression, is demonstrated to be an effective tool for gene function investigation. The aim of the present study was to observe the effect of siRNA, which was designed to target volume-regulated chloride channel ClC-2 gene, on the proliferation of a human glioma cell line U-87. Two recombinant eukaryotic CIC-2 siRNA expression vectors were designed and constructed. Sequences were identified and confirmed by restrictive endonuclease digestion and DNA sequencing. The empty vector, pSUPER. puro, and two recombinant plasmids, pSUPER. puro-shRNA-ClC-21 and pSUPER. puro-shRNA-ClC-22, were transfected into U-87 cells using Lipofectamine2000 (Groups: PP0, PP1 and PP2, respectively). ClC-2 mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR); the cellular proliferation rate was determined by MTT assay; the cell cycle was measured by flow cytometry (FCM); and the cell colony formation rate was measured by plate colony formation assay. The DNA fragments encoding siRNA targeting ClC-2-gene were successfully connected onto pSUPER. puro vector. ClC-2 mRNA expression and the cell growth rate in PP1 and PP2 groups were significantly inhibited compared to those in PP0 and control groups. Meanwhile, cell cycle was arrested in G1 phase and the percentage of G1 phase cells were increased by about 30.24% in PP1 and 18.04% in PP2 vs. in control and PP0 groups, P < 0.05. Moreover, the cell colony formation rates were statistically decreased in siRNA treated groups, which were (11.0+/-1.0)% in PP1 and (20+/-3.1)% in PP2 vs. (46.5+/-1.6)% in control and (47.5+/-2.8)% in PP0 groups (P<0.01). These results demonstrate that CIC-2 siRNA could inhibit the cell proliferation of a human glioma cell line U-87, thus ClC-2 gene may be used as a novel target for the suppression of the growth of human malignant glioma cells.


The impact of variable predation risk on stress in snowshoe hares over the cycle in North America's boreal forest: adjusting to change.
The boreal forest is one of the world's ecosystems most affected by global climate warming. The snowshoe hare, its predators, and their population dynamics dominate the mammalian component of the North American boreal forest. Our past research has shown the 9-11-year hare cycle to be predator driven, both directly as virtually all hares that die are killed by their predators, and indirectly through sublethal risk effects on hare stress physiology, behavior, and reproduction. We replicated this research over the entire cycle by measuring changes in predation risk expected to drive changes in chronic stress. We examined changes in hare condition and stress axis function using a hormonal challenge protocol in the late winter of 7 years-spanning all phases of the cycle from the increase through to the low (2014-2020). We simultaneously monitored changes in hare abundance as well as those of their primary predators, lynx and coyotes. Despite observing the expected changes in hare-predator numbers over the cycle, we did not see the predicted changes in chronic stress metrics in the peak and decline phases. Thus, the comprehensive physiological signature indicative of chronic predator-induced stress seen from our previous work was not present in this current cycle. We postulate that hares may now be increasingly showing behavior-mediated rather than stress-mediated responses to their predators. We present evidence that increases in primary productivity have affected boreal community structure and function. We speculate that climate change has caused this major shift in the indirect effects of predation on hares.


Niemann-Pick C1 functions in regulating lysosomal amine content.
Mutations in the late endosomal/lysosomal membrane protein Niemann-Pick C1 (NPC1) are known to cause a generalized block in retrograde vesicle-mediated transport, resulting in the hyper-accumulation of multiple lysosomal cargos. An important, yet often overlooked, category of lysosomal cargo includes the vast array of small molecular weight amine-containing molecules that are substrates for ion trapping in the highly acidic organelle lumen. We show here that the introduction of amine-containing molecules in lysosomes can significantly stimulate NPC1-mediated late endosome/lysosome fusion, and subsequently the secretion of lysosomal cargo. To illustrate the physiological importance of this NPC1-mediated transport pathway, we show that NPC1-deficient cells are more susceptible to the toxic effects of a lysosomotropic polyamine metabolite 3-aminopropanal. Moreover, NPC fibroblasts are shown to have higher levels of polyamine oxidase, an enzyme involved in the formation of 3-aminopropanal. Collectively, these findings provide strong support for a novel functional role for NPC1 and may also provide clues toward understanding NPC disease progression.


A Brief Talk on the Design of Biopharmaceutical Separation and Purification Technology Course.
Biological separation and purification technology is the basic technology of modern biotechnology, which is widely used in the pharmaceutical industry, especially the biopharmaceutical industry. In recent years, the biopharmaceutical industry has had a lot of room for development in the development of science and technology in South Korea, and the research on biopharmaceutical equipment and pharmaceutical technology has also achieved good research results. This article proposes a brief discussion on the design of biopharmaceutical separation and purification technology courses. In this study, by analyzing the synthesis potential of the secondary metabolites of the strain, using the α-glucosidase inhibition rate as an inspection indicator, the fermentation medium of the strain was optimized, and batch fermentation was carried out, and then, the metabolites were separated and purified, and the following conclusion was obtained: the α-glucosidase inhibition rate of the crude extract of the strain in the optimized fermentation medium is 35% higher than that of the initial medium.


Brachial plexus block as a sole anaesthetic technique in upper extremity fracture/dislocation in children: subclavian perivascular vs parascalene approach.
To compare parascalene approach of brachial plexus block with the classical subclavian perivascular approach as a sole anaesthetic technique in children undergoing closed manipulation for fracture/dislocation around the elbow. Sixty children (age 6-13 years) undergoing closed manipulation for fracture/dislocation around the elbow were randomly assigned to two groups, Group I (n=30) receiving brachial plexus block using Winnie's classical subclavian perivascular approach and Group II (n=30) receiving brachial plexus block using parascalene approach described by Dalens. Time required for performing the block, onset of analgesia, sensory block to pin prick, adequacy of relaxation, complications and acceptance of the technique to the children, parents and the surgeons were compared. Time required for performing the block (6.3+/-2.2 min vs. 8.2+/-2.4 min), onset of subjective analgesia (4.1+/-1.6 min vs. 5.2+/-1.4 min) and onset of sensory block to pinprick (6.8+/-2.1 min vs. 8.6+/-1.7 min) were significantly shorter in Group I as compared to Group II (p<0.01). Acceptance of the techniques by the children, parents and the surgeons, and the overall success rates were high and comparable between the groups. Complications were minor and the incidence was low in both the groups except Horner's syndrome in 46.7% of patients in Group II. Parascalene approach to brachial plexus block is comparable to classical subclavian perivascular approach in safety, success rate and acceptance in children undergoing closed manipulation and reduction of fracture/dislocation around the elbow.


Prevalence and psychosocial impact of urinary incontinence in older people of a Spanish rural population.
The prevalence and the psychosocial impact of urinary incontinence in older people of a rural population were investigated. A cross-sectional study was made by means of a home health interview in a representative sample of people aged 65 years and older in the Basic Health Zone of Cabra (Córdoba). A questionnaire was used with two questions to detect urinary incontinence and further questions regarding its duration, frequency, volume, severity and psychosocial impact. Data were analyzed with basic statistics. A logistic regression model was created to identify which factors affect people's lifestyles. Among the 827 respondents the prevalence of incontinence reached 36% [95% confidence interval (CI): 34%-38%] and was higher (p < .001) in women (42%; 95% CI: 39%-45%) than in men (29%; 95% CI: 25%-38%). The most frequent duration of incontinence was from 1 to 5 years (16.8%). Fifteen percent had daily episodes and 10.6% estimated the volume of urine loss as high. Incontinence was severe in 9% of the affected subjects. Half the incontinent subjects expressed social limitations and negative feelings related to incontinence, and 21% admitted that incontinence negatively affected their lives. In the logistic regression model, both volume of urine loss [odds ratio (OR) = 5.61; 95% CI: 2.76-11.42] and being married (OR = 2.34; 95% CI: 1.03-5.33) were the factors statistically associated with a negative influence of incontinence on lifestyle. Urinary incontinence is a very frequent symptom in our population. Half the incontinent subjects suffer from some degree of psychosocial limitation. The volume of urine loss and being married are the most important factors with psychosocial impact.


Influence of Pre-Existing Mild Cognitive Impairment and Dementia on Post-Stroke Mortality. The Dijon Stroke Registry.
We assessed the association between pre-stroke cognitive status and 90-day case-fatality. Patients with ischemic stroke (IS) or spontaneous intracerebral hemorrhage (ICH) were prospectively identified among residents of Dijon, France, between 2013 and 2015, using a population-based registry. Association between pre-stroke cognitive status and case-fatality at 90 days was evaluated using Cox regression. Seven hundred sixty-two patients were identified, and information about pre-stroke cognitive status was obtained for 716 (92.6%) of them, including 603 IS (84.2%) and 113 ICH (15.8%). Before stroke, 99 (13.8%) patients had mild cognitive impairment (MCI) and 98 (13.7%) had dementia. Patients with cognitive impairment were older, had a higher prevalence of several risk factors, more severe stroke, more frequent ICH, and less admission to stroke unit. Case-fatality rate at 90 days was 11.7% in patients without cognitive impairment, 32.3% in MCI patients, and 55.1% in patients with dementia. In multivariable analyses, pre-existing MCI (hazard ratio [HR] 2.22, 95% CI 1.21-4.05, p = 0.009) and dementia (HR 4.35, 95% CI 2.49-7.61, p < 0.001) were both associated with 90-day case-fatality. Pre-stroke MCI and dementia were both associated with increased mortality. These associations were not fully explained by baseline characteristics, pre-stroke dependency, stroke severity or patient management, and underlying reasons need to be investigated.


A novel long intergenic non-coding RNA, Nostrill, regulates iNOS gene transcription and neurotoxicity in microglia.
Microglia are resident immunocompetent and phagocytic cells in the CNS. Pro-inflammatory microglia, stimulated by microbial signals such as bacterial lipopolysaccharide (LPS), viral RNAs, or inflammatory cytokines, are neurotoxic and associated with pathogenesis of several neurodegenerative diseases. Long non-coding RNAs (lncRNA) are emerging as important tissue-specific regulatory molecules directing cell differentiation and functional states and may help direct proinflammatory responses of microglia. Characterization of lncRNAs upregulated in proinflammatory microglia, such as NR_126553 or 2500002B13Rik, now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus) increases our understanding of molecular mechanisms in CNS innate immunity. Microglial gene expression array analyses and qRT-PCR were used to identify a novel long intergenic non-coding RNA, Nostrill, upregulated in LPS-stimulated microglial cell lines, LPS-stimulated primary microglia, and LPS-injected mouse cortical tissue. Silencing and overexpression studies, RNA immunoprecipitation, chromatin immunoprecipitation, chromatin isolation by RNA purification assays, and qRT-PCR were used to study the function of this long non-coding RNA in microglia. In vitro assays were used to examine the effects of silencing the novel long non-coding RNA in LPS-stimulated microglia on neurotoxicity. We report here characterization of intergenic lncRNA, NR_126553, or 2500002B13Rik now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus). Nostrill is induced by LPS stimulation in BV2 cells, primary murine microglia, and in cortical tissue of LPS-injected mice. Induction of Nostrill is NF-κB dependent and silencing of Nostrill decreased inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in BV2 and primary microglial cells. Overexpression of Nostrill increased iNOS expression and NO production. RNA immunoprecipitation assays demonstrated that Nostrill is physically associated with NF-κB subunit p65 following LPS stimulation. Silencing of Nostrill significantly reduced NF-κB p65 and RNA polymerase II recruitment to the iNOS promoter and decreased H3K4me3 activating histone modifications at iNOS gene loci. In vitro studies demonstrated that silencing of Nostrill in microglia reduced LPS-stimulated microglial neurotoxicity. Our data indicate a new regulatory role of the NF-κB-induced Nostrill and suggest that Nostrill acts as a co-activator of transcription of iNOS resulting in the production of nitric oxide by microglia through modulation of epigenetic chromatin remodeling. Nostrill may be a target for reducing the neurotoxicity associated with iNOS-mediated inflammatory processes in microglia during neurodegeneration.


homeSound: Real-Time Audio Event Detection Based on High Performance Computing for Behaviour and Surveillance Remote Monitoring.
The consistent growth in human life expectancy during the recent years has driven governments and private organizations to increase the efforts in caring for the eldest segment of the population. These institutions have built hospitals and retirement homes that have been rapidly overfilled, making their associated maintenance and operating costs prohibitive. The latest advances in technology and communications envisage new ways to monitor those people with special needs at their own home, increasing their quality of life in a cost-affordable way. The purpose of this paper is to present an Ambient Assisted Living (AAL) platform able to analyze, identify, and detect specific acoustic events happening in daily life environments, which enables the medic staff to remotely track the status of every patient in real-time. Additionally, this tele-care proposal is validated through a proof-of-concept experiment that takes benefit of the capabilities of the NVIDIA Graphical Processing Unit running on a Jetson TK1 board to locally detect acoustic events. Conducted experiments demonstrate the feasibility of this approach by reaching an overall accuracy of 82% when identifying a set of 14 indoor environment events related to the domestic surveillance and patients' behaviour monitoring field. Obtained results encourage practitioners to keep working in this direction, and enable health care providers to remotely track the status of their patients in real-time with non-invasive methods.


The effect of chronic corneal epithelial debridement on epithelial and stromal morphology in dogs.
To determine the effect of chronic corneal epithelial debridement on epithelial and stromal morphology and extracellular matrix components, and to compare those changes to those in spontaneous chronic corneal epithelial defects (SCCED) in dogs. Axial corneal epithelial wounds, 10 mm in diameter, were created weekly for 8 weeks in five normal adult laboratory beagles. Slit lamp biomicroscopy and corneal pachymetry were performed weekly before wounding. Three days after the last debridement the dogs were killed humanely, and corneas were processed for light and electron microscopy and immunohistochemistry for collagen IV, collagen VII, fibronectin, and laminin. No significant changes in corneal thickness were found. All samples demonstrated epithelial dysmaturation adjacent to the wound edge, and, in four of five, a narrow zone of nonadherent epithelium formed adjacent to the exposed stroma. All samples had a stromal acellular zone in the area of the defect and continuing for a short distance under the adjacent attached epithelium. Experimentally wounded dogs did not form the superficial hyaline acellular lamina found in 92% of dogs with SCCED. Laminin, collagen IV, and fibronectin were present on the stromal surface in all samples, and collagen VII was present in four of five samples. Transmission electron microscopy (TEM) demonstrated the presence of basement membrane on the surface of the exposed stroma. Epithelial changes are similar between experimentally wounded dogs and dogs with SCCED. The stromal acellular zone that forms in experimentally wounded dogs is distinct from the hyaline lamina observed in dogs with SCCED. The difference in the acellular stromal layers between chronically wounded dogs and dogs with SCCED may be of relevance to our understanding of the pathophysiology of persistent epithelial defects.


Identification and Optimization of Novel Inhibitors of the Polyketide Synthase 13 Thioesterase Domain with Antitubercular Activity.
There is an urgent need for new tuberculosis (TB) treatments, with novel modes of action, to reduce the incidence/mortality of TB and to combat resistance to current treatments. Through both chemical and genetic methodologies, polyketide synthase 13 (Pks13) has been validated as essential for mycobacterial survival and as an attractive target for Mycobacterium tuberculosis growth inhibitors. A benzofuran series of inhibitors that targeted the Pks13 thioesterase domain, failed to progress to preclinical development due to concerns over cardiotoxicity. Herein, we report the identification of a novel oxadiazole series of Pks13 inhibitors, derived from a high-throughput screening hit and structure-guided optimization. This new series binds in the Pks13 thioesterase domain, with a distinct binding mode compared to the benzofuran series. Through iterative rounds of design, assisted by structural information, lead compounds were identified with improved antitubercular potencies (MIC < 1 μM) and in vitro ADMET profiles.


[Application of a tumor cell vaccine transfected with GM-CSF/IL-2 fusion gene for specific immunotherapy of hepatocellular carcinoma].
To prepare a transgenic tumor cell vaccine transfected the fusion gene of murine granulocyte-monocyte colony stimulating factor (mGM-CSF) and human interleukin-2 (hIL-2) using H22 cells, and explore its specific antitumor immunity against hepatocellular carcinoma. The eukaryotic vector expressing the fusion gene mGM-CSF/hIL-2 was transfected into H22 cells followed by radiation exposure to construct the tumor cell vaccine, which was used to immunize Balb/c mice by subcutaneous inoculation. The mice inoculated subcutaneously with H22 cells, cells transfected with the empty vector pcDNA(+), or with PBS served as the controls. A week later, H22 cells were injected peritoneally into Balb/c mice for establishing the tumor-bearing model, and their serum levels of interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) were detected using enzyme-linked immunosorbent assay (ELISA) with the survival of the mice recorded. The spleen cells were obtained from the mice immunized with the tumor cell vaccine, the tumor-bearing mice and the normal control mice to assess their cytotoxicity against the parental H22 cells in vitro using (51)C(r)-release assay. The transgenic H22 cell vaccine transfected with mGM-CSF/hIL-2 fusion gene was successfully constructed. The killing rate of H22 cells by the spleen cells from the mice immunized with the transgenic cell vaccine was significantly higher than those by the spleen cells from the tumor-bearing mice or normal control mice (38.3% vs 13.6% and 7.5%, P<0.05). Serum IFN-gamma in the tumor-bearing mice immunized with the transgenic cell vaccine was significantly higher, and serum IL-10 significantly lower than those of the control groups (P<0.01). The survival time of the tumor-bearing mice injected with the transgenic cell vaccine was also significantly prolonged. Syngeneic tumor cell vaccine genetically modified by mGM-CSF/hIL-2 fusion gene transfection can elicit specific cellular immune response and enhance the host antitumor immune response to extend the survival time of tumor-bearing mice.


Cellular interactions of synovial fluid γδ T cells in juvenile idiopathic arthritis.
The pathogenesis of juvenile idiopathic arthritis (JIA) is thought to involve multiple components of the cellular immune system, including subsets of γδ T cells. In this study, we conducted experiments to define the functional roles of one of the major synovial fluid (SF) T cell subsets, Vγ9(+)Vδ2(+) (Vγ9(+)) T cells, in JIA. We found that as opposed to CD4(+) T cells, equally high percentages (∼35%) of Vγ9(+) T cells in SF and peripheral blood (PB) produced TNF-α and IFN-γ. Furthermore, stimulation with isopentenyl pyrophosphate (IPP), a metabolite in the mevalonate pathway, which is a specific potent Ag for Vγ9Jγ1.2(+) T cells, similarly amplified cytokine secretion by SF and PB Vγ9(+) T cells. Significantly, the SF subset expressed higher levels of CD69 in situ, suggesting their recent activation. Furthermore, 24-h coculturing with SF-derived fibroblasts enhanced CD69 on the SF > PB Vγ9(+) T cells, a phenomenon strongly augmented by zoledronate, a farnesyl pyrophosphate synthase inhibitor that increases endogenous intracellular IPP. Importantly, although Vγ9(+) T cell proliferation in response to IPP was significantly lower in SF than PBMC cultures, it could be enhanced by depleting SF CD4(+)CD25(+)FOXP3(+) cells (regulatory T cells). Furthermore, coculture with the Vγ9(+) T cells in medium containing zoledronate or IPP strongly increased SF-derived fibroblasts' apoptosis. The findings that IPP-responsive proinflammatory synovial Vγ9(+) T cells for which proliferation is partly controlled by regulatory T cells can recognize and become activated by SF fibroblasts and then induce their apoptosis suggest their crucial role in the pathogenesis and control of synovial inflammation.


Characteristics and time points to inhibit ferroptosis in human osteoarthritis.
Ferroptosis is a form of cell death that is triggered by iron-dependent lipid peroxidation and is closely associated with osteoarthritis. The primary interventions for inhibiting ferroptosis in osteoarthritis are anti-lipid peroxidation and iron chelation. The objective of our study is to investigate the characteristics of ferroptosis in osteoarthritis and identify the optimal time points for inhibiting ferroptosis to alleviate disease progression. Ferroptosis-related alterations and markers of OA were analyzed in paired intact and damaged cartilages from OA patients by immunofluorescence, qRT-PCR, mitochondrial membrane potential and immunohistochemistry. We also compared Ferroptosis-related alterations in cartilage of mild, moderate, and severe OA (according to the modified Mankin score). In addition, we compared the effect of Fer-1 on ferroptosis and the protection of chondrocytes by detecting markers of both ferroptosis and OA by immunofluorescence, CCK8 and qRT-PCR. Ferroptosis-related alterations (GPX4 downregulation, ACSL4 upregulation, MDA, LPO accumulation, Mitochondrial membrane potential decreased) in the damaged area cartilage were more severe than those in the intact area and increased with the progression of OA. Compared with mild OA group, the activity of chondrocytes treated with Fer-1 (a ferroptosis inhibitor) was increased, mitochondrial function was improved, and ferroptosis was reduced (GPX4 upregulation, SLC7A11 upregulation, ACSL4 downregulation,), and promoted the expression of COL2A1 and inhibited the expression of MMP13. However, these changes were not observed in moderate and severe OA chondrocytes. Ferroptosis occurs in a region-specific manner and is exacerbated with the progression of human OA cartilage degeneration. Inhibition of ferroptosis might had a therapeutic effect on chondrocytes with mild OA but had no significant therapeutic effect on chondrocytes with moderate to severe OA.


Effects of dependence in high-dimensional multiple testing problems.
We consider effects of dependence among variables of high-dimensional data in multiple hypothesis testing problems, in particular the False Discovery Rate (FDR) control procedures. Recent simulation studies consider only simple correlation structures among variables, which is hardly inspired by real data features. Our aim is to systematically study effects of several network features like sparsity and correlation strength by imposing dependence structures among variables using random correlation matrices. We study the robustness against dependence of several FDR procedures that are popular in microarray studies, such as Benjamin-Hochberg FDR, Storey's q-value, SAM and resampling based FDR procedures. False Non-discovery Rates and estimates of the number of null hypotheses are computed from those methods and compared. Our simulation study shows that methods such as SAM and the q-value do not adequately control the FDR to the level claimed under dependence conditions. On the other hand, the adaptive Benjamini-Hochberg procedure seems to be most robust while remaining conservative. Finally, the estimates of the number of true null hypotheses under various dependence conditions are variable. We discuss a new method for efficient guided simulation of dependent data, which satisfy imposed network constraints as conditional independence structures. Our simulation set-up allows for a structural study of the effect of dependencies on multiple testing criterions and is useful for testing a potentially new method on pi0 or FDR estimation in a dependency context.


Morphological and Molecular Species Identification of Termites Attacking Ironwood Trees, Casuarina equisetifolia (Fagales: Casuarinaceae), in Guam.
Ironwood trees (Casuarina equisetifolia subsp. equisetifolia L.) are ecologically and economically important trees in tropical and subtropical regions of the Indo-Pacific. Ironwood is one of the dominant tree species in Guam, but since 2002, this tree has been declining dramatically. A previous study showed that numerous sick or dead trees were under termite attack. However, the species of termites were not identified. As a first step to investigate causal relationships between termites and ironwood tree death, we assigned termites collected from ironwood trees to species using a combination of morphological characters and DNA barcoding of the 12S, 16S, COI, COII, and ITS2 regions. Based on morphology and comparisons to reference sequences in NCBI GenBank, the most likely species assignments were Nasutitermes takasagoensis (Nawa) (Blattodea: Termitidae) found to infest 45 trees, followed by Coptotermes gestroi (Wasmann) (Blattodea: Rhinotermitidae) (2 trees), Microcerotermes crassus Snyder (Blattodea: Termitidae) (2 trees), and an additional unidentified Microcerotermes species (1 tree) with no close sequence match to identified species in NCBI GenBank. However, taxonomic revisions and broader representation of DNA markers of well-curated specimen in public databases are clearly needed, especially for the N. takasagoensis species complex.


The impact of machine learning on patient care: A systematic review.
Despite the expanding use of machine learning (ML) in fields such as finance and marketing, its application in the daily practice of clinical medicine is almost non-existent. In this systematic review, we describe the various areas within clinical medicine that have applied the use of ML to improve patient care. A systematic review was performed in accordance with the PRISMA guidelines using Medline(R), EBM Reviews, Embase, Psych Info, and Cochrane Databases, focusing on human studies that used ML to directly address a clinical problem. Included studies were published from January 1, 2000 to May 1, 2018 and provided metrics on the performance of the utilized ML tool. A total of 1909 unique publications were reviewed, with 378 retrospective articles and 8 prospective articles meeting inclusion criteria. Retrospective publications were found to be increasing in frequency, with 61 % of articles published within the last 4 years. Prospective articles comprised only 2 % of the articles meeting our inclusion criteria. These studies utilized a prospective cohort design with an average sample size of 531. The majority of literature describing the use of ML in clinical medicine is retrospective in nature and often outlines proof-of-concept approaches to impact patient care. We postulate that identifying and overcoming key translational barriers, including real-time access to clinical data, data security, physician approval of "black box" generated results, and performance evaluation will allow for a fundamental shift in medical practice, where specialized tools will aid the healthcare team in providing better patient care.


Cholesterol accumulation in macrophages drives NETosis in atherosclerotic plaques via IL-1β secretion.
Neutrophil extracellular trap formation (NETosis) increases atherosclerotic plaque vulnerability and athero-thrombosis. However, mechanisms promoting NETosis during atherogenesis are poorly understood. We have shown that cholesterol accumulation due to myeloid cell deficiency of the cholesterol transporters ATP Binding Cassette A1 and G1 (ABCA1/G1) promotes NLRP3 inflammasome activation in macrophages and neutrophils and induces prominent NETosis in atherosclerotic plaques. We investigated whether NETosis is a cell-intrinsic effect in neutrophils or is mediated indirectly by cellular crosstalk from macrophages to neutrophils involving IL-1β. We generated mice with neutrophil or macrophage-specific Abca1/g1 deficiency (S100A8CreAbca1fl/flAbcg1fl/fl or CX3CR1CreAbca1fl/flAbcg1fl/fl mice, respectively), and transplanted their bone marrow into low-density lipoprotein receptor knockout mice. We then fed the mice a cholesterol-rich diet. Macrophage, but not neutrophil Abca1/g1 deficiency activated inflammasomes in macrophages and neutrophils, reflected by caspase-1 cleavage, and induced NETosis in plaques. NETosis was suppressed by administering an interleukin (IL)-1β neutralizing antibody. The extent of NETosis in plaques correlated strongly with the degree of neutrophil accumulation, irrespective of blood neutrophil counts, and neutrophil accumulation was decreased by IL-1β antagonism. In vitro, IL-1β or media transferred from Abca1/g1-deficient macrophages increased NETosis in both control and Abca1/Abcg1 deficient neutrophils. This cell-extrinsic effect of IL-1β on NETosis was blocked by an NLRP3 inhibitor. These studies establish a new link between inflammasome-mediated IL-1β production in macrophages and NETosis in atherosclerotic plaques. Macrophage-derived IL-1β appears to increase NETosis both by increasing neutrophil recruitment to plaques and by promoting neutrophil NLRP3 inflammasome activation.


Structural mechanism governing cis and trans isomeric states and an intramolecular switch for cis/trans isomerization of a non-proline peptide bond observed in crystal structures of scorpion toxins.
Non-proline cis peptide bonds have been observed in numerous protein crystal structures even though the energetic barrier to this conformation is significant and no non-prolyl-cis/trans-isomerase has been identified to date. While some external factors, such as metal binding or co-factor interaction, have been identified that appear to induce cis/trans isomerization of non-proline peptide bonds, the intrinsic structural basis for their existence and the mechanism governing cis/trans isomerization in proteins remains poorly understood. Here, we report the crystal structure of a newly isolated neurotoxin, the scorpion alpha-like toxin Buthus martensii Karsch (BmK) M7, at 1.4A resolution. BmK M7 crystallizes as a dimer in which the identical non-proline peptide bond between residues 9 and 10 exists either in the cis conformation or as a mixture of cis and trans conformations in either monomer. We also determined the crystal structures of several mutants of BmK M1, a representative scorpion alpha-like toxin that contains an identical non-proline cis peptide bond as that observed in BmK M7, in which residues within or neighboring the cis peptide bond were altered. Substitution of an aspartic acid residue for lysine at residue 8 in the BmK M1 (K8D) mutant converted the cis form of the non-proline peptide bond 9-10 into the trans form, revealing an intramolecular switch for cis-to-trans isomerization. Cis/trans interconversion of the switch residue at position 8 appears to be sequence-dependent as the peptide bond between residues 9 and 10 retains its wild-type cis conformation in the BmK M1 (K8Q) mutant structure. The structural interconversion of the isomeric states of the BmK M1 non-proline cis peptide bond may relate to the conversion of the scorpion alpha-toxins subgroups.


Enhanced hypertension care through private clinics in Pakistan: a cluster randomised trial.
Hypertension in Pakistan affects 33% of people aged ≥45 years, and in urban areas around 70% of basic health care occurs in private facilities. To assess whether enhanced care at urban private clinics resulted in better control of hypertension, cardiovascular disease (CVD) risk factors, and treatment adherence. A two-arm cluster randomised controlled trial was conducted at 26 private clinics (in three districts of Punjab) between January 2015-September 2016. Both arms had enhanced screening and diagnosis of hypertension and related conditions, and patient recording processes. Intervention facilities also had a clinical care guide, additional drugs for hypertension, a patient lifestyle education flipchart, associated training, and mobile phone follow-up. Clinics were randomised in a 1:1 ratio (sealed envelope lottery method). A total of 574 intervention and 564 control patients in 13 clusters in each arm were recruited (male and female, aged ≥25 years, systolic blood pressure [SBP] >140 mmHg, and/or diastolic blood pressure [DBP] >90 mmHg). The primary outcome was change in SBP from baseline to 9-month follow-up.Staff and patients were not blinded, but outcome assessors were blinded. Nine-month primary outcomes were available for 522/574 (90.9%) intervention and 484/564 (85.8%) control participants (all clusters). The unadjusted cluster-level analysis results were as follows: mean intervention outcome was -25.2 mmHg (95% confidence intervals [CI] = -29.9 to-20.6); mean control outcome was -9.4 mmHg (95% CI = 21.2 to 2.2); and mean control-intervention difference was 15.8 (95% CI = 3.6 to 28.0; P = 0.01). The findings and separate process evaluation support the scaling of an integrated CVD-hypertension care intervention in urban private clinics in areas lacking public primary care in Pakistan.


Abnormal polarity of thunderclouds grown from negatively charged air.
Experiments were carried out in New Mexico to determine whether the electrification processes that lead to the formation of lightning in clouds are influenced by the polarity of the charges in the air from which the clouds grow. The normal, positive space charge in the sub-cloud air was reversed by negative charge released from an electrified wire, suspended across a 2-kilometer-wide canyon. On more than four occasions when the clouds over the wire grew and became electrified, they were of abnormal polarity with dominant positive charges instead of the usual negative charges in the lower part of the cloud. The formation of these abnormally electrified clouds suggests both that the electrification process in thunderclouds can be initiated and that its polarity may be determined by the small charges that are present in the atmosphere.


A mixed treatment comparison of selected osteopathic techniques used to treat acute nonspecific low back pain: a proof of concept and plan for further research.
Back injuries have a high prevalence in the United States and can be costly for both patients and the healthcare system at large. While previous guidelines from the American College of Physicians for the management of acute nonspecific low back pain (ANLBP) have encouraged nonpharmacologic management, those treatment recommendations involved only superficial heat, massage, acupuncture, and spinal manipulation. Investigation about the efficacy of spinal manipulation in the management of ANLBP is warranted. To compare the results in previously-published literature documenting the outcomes of osteopathic manipulative treatment (OMT) techniques used to treat ANLBP. The secondary objective of this study was to demonstrate the utility of using Bayesian network meta-analysis (NMA) to perform a mixed treatment comparison (MTC) of a variety of osteopathic techniques. A literature search for randomized controlled trials (RCTs) of ANLBP treatments was performed in April 2020 according to PRISMA guidelines by searching MEDLINE/PubMed, OVID, Cochrane Central, PEDro, and OSTMED.Dr databases; scanning the reference lists of articles; and using the Canadian Agency for Drugs and Technologies in Health grey literature checklist. Each database was searched from inception to April 1, 2020. The following search terms were used: acute low back pain, acute low back pain plus physical therapy, acute low back pain plus spinal manipulation, and acute low back pain plus osteopathic manipulation. The validity of eligible trials was assessed by the single author using an adapted National Institute for Health and Care Excellence methodology checklist for randomized, controlled trials and an extraction form based on that checklist. The outcome measure chosen for this NMA was the Visual Analogue Scale of pain. The NMA were performed using the GeMTC user interface for automated NMA utilizing a Bayesian hierarchical model of random effects. The literature search initially found 483 unduplicated records. After screening and full text assessment, five RCTs were eligible for the MTC, yielding a total of 430 participants. Results of the MTC model suggested that there was no statistically significant decrease in reported pain when exercise, high-velocity low-amplitude (HVLA), counterstrain, muscle energy technique, or a mix of techniques were added to conventional treatment to treat ANLBP. However, the rank probabilities assessment determined that HVLA and the OMT mixed treatment protocol plus conventional care were ranked superior to conventional care alone for improving ANLBP. While this study failed to provide definitive evidence upon which clinical recommendations can be based, it does demonstrate the utility of performing NMA for MTCs of osteopathic modalities used to treat ANLBP. However, to take full advantage of this statistical technique, future studies should be designed with consideration for the methodological shortcomings found in past osteopathic research.


Synergy of photocatalysis and fuel cells: A chronological review on efficient designs, potential materials and emerging applications.
The rising demand of energy and lack of clean water are two major concerns of modern world. Renewable energy sources are the only way out in order to provide energy in a sustainable manner for the ever-increasing demands of the society. A renewable energy source which can also provide clean water will be of immense interest and that is where Photocatalytic Fuel Cells (PFCs) exactly fit in. PFCs hold the ability to produce electric power with simultaneous photocatalytic degradation of pollutants on exposure to light. Different strategies, including conventional Photoelectrochemical cell design, have been technically upgraded to exploit the advantage of PFCs and to widen their applicability. Parallel to the research on design, researchers have put an immense effort into developing materials/composites for electrodes and their unique properties. The efficient strategies and potential materials have opened up a new horizon of applications for PFCs. Recent research reports reveal this persistently broadening arena which includes hydrogen and hydrogen peroxide generation, carbon dioxide and heavy metal reduction and even sensor applications. The review reported here consolidates all the aspects of various design strategies, materials and applications of PFCs. The review provides an overall understanding of PFC systems, which possess the potential to be a marvellous renewable source of energy with a handful of simultaneous applications. The review is a read to the scientific community and early researchers interested in working on PFC systems.


Robust transformation procedure for the production of transgenic farmer-preferred cassava landraces.
Recent progress in cassava transformation has allowed the robust production of transgenic cassava even under suboptimal plant tissue culture conditions. The transformation protocol has so far been used mostly for the cassava model cultivar 60444 because of its good regeneration capacity of embryogenic tissues. However, for deployment and adoption of transgenic cassava in the field it is important to develop robust transformation methods for farmer- and industry-preferred landraces and cultivars. Because dynamics of multiplication and regeneration of embryogenic tissues differ between cassava genotypes, it was necessary to adapt the efficient cv. 60444 transformation protocol to genotypes that are more recalcitrant to transformation. Here we demonstrate that an improved cassava transformation protocol for cv. 60444 could be successfully modified for production of transgenic farmer-preferred cassava landraces. The modified transformation method reports on procedures for optimization and is likely transferable to other cassava genotypes reportedly recalcitrant to transformation provided production of high quality FEC. Because the three farmer-preferred cassava landraces selected in this study have been identified as resistant or tolerant to cassava mosaic disease (CMD), the adapted protocol will be essential to mobilize improved traits into cassava genotypes suitable for regions where CMD limits production.


Development of an in vitro carcinogenesis model of human papillomavirus-induced cervical adenocarcinoma.
Cervical adenocarcinoma (ADC) is the second most common pathological subtype of cervical cancer after squamous cell carcinoma. It accounts for approximately 20% of cervical cancers, and the incidence has increased in the past few decades, particularly among young patients. The persistent infection of high-risk human papillomavirus (HPV) is responsible for most cervical ADC. However, almost all available in vitro models are designed to study the carcinogenesis of cervical squamous cell carcinoma. To gain better insights into molecular background of ADC, we aimed to establish an in vitro carcinogenesis model of ADC. We previously reported the establishment of an in vitro model for cervical squamous cell carcinoma by introducing defined viral and cellular oncogenes, HPV16 E6 and E7, c-MYC, and activated RAS to human cervical keratinocytes. In this study, the expression of potential lineage-specifying factors and/or SMAD4 reduction was introduced in addition to the defined four oncogenes to direct carcinogenesis toward ADC. The cell properties associated with the cell lineage were analyzed in monolayer and organoid cultures and the tumors in mouse xenografts. In the cells expressing Forkhead box A2 (FOXA2), apparent changes in cell properties were observed, such as elevated expression of columnar cell markers and decreased expression of squamous cell markers. Strikingly, the histopathology of tumors expressing FOXA2 resembled cervical ADC, proposing that FOXA2 plays a vital role in dictating the histopathology of cervical cancers.


Accessing seasonal weather forecasts and drought prediction information for rural households in Chirumhanzu district, Zimbabwe.
Seasonal weather forecasts and drought hazard prediction through media sources and indigenous knowledge help provide an understanding of early warning systems and the preferred source information by rural households. This article focuses on the investigation of households' access to weather forecasts and drought hazard prediction information as early warning to reduce drought risk on livelihood activities. The study was carried out in Chirumhanzu district, and the methods used for data collection included 217 household surveys, six focus group discussions, key informants' interviews and document review. The study found that the majority of the households in the study area had access to seasonal weather forecast information (scientific), which almost half of the respondents received through radios. However, vulnerability to climate risks was exacerbated by seasonal weather forecasts, which were deemed by some households to be unreliable, inaccurate and not easily understood. In this regard, some households used indigenous knowledge to inform them on the status of the incoming rainy season and drought prediction. The use of indigenous knowledge depended on individuals' ability to read and decode natural indicators of seasonal weather forecast and drought prediction. Indigenous knowledge is valuable for climate science as it enhances observations and interpretations on a larger spatial scale with considerable temporal depth by highlighting elements that are measured by climate science. Both scientific weather information and indigenous knowledge are important for seasonal weather forecasting and drought prediction, especially in rural settings, and complement each other if used and availed timely to households.


Physical activity level and its association with metabolic syndrome among an employed population in China.
The purpose of the present study was to estimate the association of physical activity level (PAL) with metabolic syndromes among the employed population in China. Using data from the 2002 China National Nutrition and Health Survey, the PAL of 21 834 subjects aged 18-59 years was described, and its relationship with the risk of metabolic syndrome was studied after adjusting for the relative effects of smoking, drinking, diet and socioeconomic status. Information on physical activity was collected by trained investigators using a 1-year physical activity questionnaire. PAL was classified into four categories: sedentary (PAL 1.00-1.39), low active (PAL 1.40-1.59), active (PAL 1.60-1.89) and very active (PAL 1.90-2.50). The definition of metabolic syndrome by the International Diabetes Federation 2005 was applied. The percentage of people being sedentary, low active, active and very active was 12.3%, 13.7%, 20.1% and 53.9% respectively. The crude prevalence of metabolic syndrome in the sedentary, low active, active and very active groups was 9.7%, 6.9%, 5.6% and 4.9% respectively. After adjusting for the effect of other risk factors, the higher the PAL, the lower the relative risk of metabolic syndrome as well as the individual metabolic abnormalities. The risk of overweight/obesity and metabolic syndrome was 50% and 59% less, respectively, in the adults who were very active and whose dietary fat contributed less than 20% to energy intakes when compared with their counterparts who were sedentary and consumed more fat (> or = 30% energy). PAL was an independent risk factor for obesity and multiple metabolic syndrome abnormalities among the employed population in China. The high-risk population were middle-age males living in large cities with high incomes who drink alcohol.


Intrinsic morphological diversity of thick-tufted layer 5 pyramidal neurons ensures robust and invariant properties of in silico synaptic connections.
The morphology of neocortical pyramidal neurons is not only highly characteristic but also displays an intrinsic diversity that renders each neuron morphologically unique. We investigated the significance of this intrinsic morphological diversity in in silico networks composed of thick-tufted layer 5 (TTL5) pyramidal neurons, by comparing the in silico and in vitro properties of TTL5 synaptic connections. The synaptic locations of in silico connections were determined by placing 3D reconstructed TTL5 neurons randomly in a volume equivalent to that of layer 5 in the juvenile rat somatosensory cortex and using a 'collision-detection' algorithm to identify the incidental loci of axo-dendritic overlap. The activation time of the modelled synapses and their biophysical properties were characterized based on experimental measurements. We found that the anatomical loci of synapses and the physiological properties of the somatically recorded EPSPs closely matched those recorded experimentally without the need for any fine-tuning. Furthermore, perturbations to both the physiological or anatomical parameters of the model did not alter the average physiological properties of the population of modelled synaptic connections. This microcircuit-level robust behaviour was due to the intrinsic diversity of the morphology of pyramidal neurons in the microcircuit. We conclude that synaptic transmission in a network of TTL5 neurons is highly invariant across microcircuits suggesting that intrinsic diversity is a mechanism to ensure the same average synaptic properties in different animals of the same species. Finally, we show that the average physiological properties of the TTL5 microcircuit are surprisingly robust to anatomical and physiological perturbations also partly due to the intrinsic diversity of pyramidal neuron morphology.


Grip Strength: An Indispensable Biomarker For Older Adults.
Grip strength has been proposed as a biomarker. Supporting this proposition, evidence is provided herein that shows grip strength is largely consistent as an explanator of concurrent overall strength, upper limb function, bone mineral density, fractures, falls, malnutrition, cognitive impairment, depression, sleep problems, diabetes, multimorbidity, and quality of life. Evidence is also provided for a predictive link between grip strength and all-cause and disease-specific mortality, future function, bone mineral density, fractures, cognition and depression, and problems associated with hospitalization. Consequently, the routine use of grip strength can be recommended as a stand-alone measurement or as a component of a small battery of measurements for identifying older adults at risk of poor health status.


Characterization of tissue culture-induced heterogeneity in DNAs of independent isolates of JC virus.
After several serial passages at low multiplicities of infection in primary human foetal glial cells at 37 degrees C, the DNA of prototype (MAD-1) JC virus and that of MAD-2 and MAD-3 are typically heterogeneous in size, but DNAs of MAD-4 and MAD-6 are relatively homogeneous. A similar dichotomy was observed in the DNAs of six isolates propagated more recently in glial cultures at 39 degrees C under similar conditions of brief passage in vitro at low multiplicities of infection: the DNAs of two (MAD-9 and -10) were heterogeneous, but the DNAs of four others (MAD-8, -11, -12 and -14) were homogeneous. Therefore, the propensity of the viral genome to sustain deletions was an intrinsic property of each isolate. However, actual induction and maintenance of the presumably defective DNAs was influenced by the relative proportions of permissive spongioblasts and semi-permissive astrocytes in the glial cultures and by the multiplicity of infection. Deletions in MAD-1 DNA were confined to the presumptive early region and spanned the BamHI cleavage site (map position 0.505). The heterogeneity was more complex in the DNAs of MAD-2 and MAD-3, but again most of the deletions, which ranged up to 12% of full-length DNA, spanned the BamHI site. We propose that the differential susceptibility to deletion among isolates is a consequence of natural genetic variation in JC virus.


TMEM43 mutations associated with arrhythmogenic right ventricular cardiomyopathy in non-Newfoundland populations.
Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a myocardial disease characterized by fibro-fatty replacement of right ventricular free wall myocardium and life-threatening ventricular arrhythmias. A missense mutation, c.1073C>T (p.S358L) in the transmembrane protein 43 (TMEM43) gene, has been genetically identified to cause ARVC type 5 in a founder population from Newfoundland. It is unclear whether this mutation occurs in other populations outside of this founder population or if other variants of TMEM43 are associated with ARVC disease. We sought to identify non-Newfoundland individuals with TMEM43 variants among patient samples sent for genetic assessment for possible ARVC. Of 195 unrelated individuals with suspected ARVC, mutation of desmosomal proteins was seen in 28 and the p.S358L TMEM43 mutation in six. We identified a de novo p.S358L mutation in a non-Newfoundland patient and five separate rare TMEM43 (four novel) sequence variants in non-Newfoundland patients, each occurring in an evolutionarily conserved amino acid. TMEM43 mutations occur outside of the founder population of the island of Newfoundland where it was originally described. TMEM43 sequencing should be incorporated into clinical genetic testing for ARVC patients.


Corroboration of mechanoregulatory algorithms for tissue differentiation during fracture healing: Comparison with in vivo results.
Several mechanoregulation algorithms proposed to control tissue differentiation during bone healing have been shown to accurately predict temporal and spatial tissue distributions during normal fracture healing. As these algorithms are different in nature and biophysical parameters, it raises the question of which reflects the actual mechanobiological processes the best. The aim of this study was to resolve this issue by corroborating the mechanoregulatory algorithms with more extensive in vivo bone healing data from animal experiments. A poroelastic three-dimensional finite element model of an ovine tibia with a 2.4 mm gap and external callus was used to simulate the course of tissue differentiation during fracture healing in an adaptive model. The mechanical conditions applied were similar to those used experimentally, with axial compression or torsional rotation as two distinct cases. Histological data at 4 and 8 weeks, and weekly radiographs, were used for comparison. By applying new mechanical conditions, torsional rotation, the predictions of the algorithms were distinguished successfully. In torsion, the algorithms regulated by strain and hydrostatic pressure failed to predict healing and bone formation as seen in experimental data. The algorithm regulated by deviatoric strain and fluid velocity predicted bridging and healing in torsion, as observed in vivo. The predictions of the algorithm regulated by deviatoric strain alone did not agree with in vivo data. None of the algorithms predicted patterns of healing entirely similar to those observed experimentally for both loading modes. However, patterns predicted by the algorithm based on deviatoric strain and fluid velocity was closest to experimental results. It was the only algorithm able to predict healing with torsional loading as seen in vivo.


Coevolution with higher taxonomic host groups within the Puccinia/Uromyces rust lineage obscured by host jumps.
Partial beta-tubulin 1 sequence data were obtained for 80 taxa of Pucciniaceae, with hosts from 33 angiosperm families, covering all major ordinal groups in the Angiosperm Phylogeny Group classification. As in previous studies, most species of Puccinia and Uromyces fell into two main clades (I and II), with P. glechomatis and P. psidii excluded from Pucciniaceae. Results suggest two processes; a coevolution of and hosts in each clade, as well as associated frequent jumps to ecologically close, but taxonomically distant, hosts. Clade I contained all rusts on Cyperaceae and Juncaceae, and most rusts on host orders from rosids to euasterids II. Clade II contained all rusts on Poaceae, and most on host orders from monocots to core eudicots. In both main clades, several well-supported subclades were identified. The grouping in clade I, subclade E of rusts of Cyperaceae and Asteraceae and, in particular, of an Australian isolate of P. dioicae with rusts on Australian families of Asterales, suggested a local radiation, and supported the coevolutionary relationship between rusts on these two families seen with a different range of asteraceous rusts in the Northern Hemisphere. In clade I, two clades contained only rusts of Asteraceae and Fabaceae, respectively, and in clade II, subclade F contained only rusts of pooid hosts. Rusts on non-pooid hosts were separated from them in subclade G. Other subclades contained a range of rusts on distantly related angiosperm families. Urediniospore morphology was often, but not always, correlated with the molecular phylogeny. Most rusts with urediniospores having few (1-5) equatorial germ pores were in clade I, whereas most with spores having several (5-14) scattered pores were in clade II. The distribution of telial host families on the beta-tubulin rust phylogeny was not random. Aecial hosts of heteroecious rusts played an important role in the evolutionary process. Possible examples of host jumps were seen in rusts on Geraniaceae, Polygonaceae, and Apiaceae. Despite such jumps obscuring past host associations, possible ancestral hosts were identified by the pattern of host distribution at higher taxonomic levels along the ss-tubulin phylogeny. Results suggest that clade I diverged with Cyperaceae, Juncaceae, and the more advanced core eudicot orders (rosids and asterids), whereas clade II diversified with earlier angiosperm groups, such as monocots, Poaceae, and Ranunculales. Qualified support was given to the hypothesis that rusts can reveal taxonomic relationships between their hosts, at genus, family, and ordinal levels.


Cellular and molecular mechanisms of dendrite growth.
Proper growth and branching of dendrites are crucial for nervous system function; patterns of dendritic arborization determine the nature and amount of innervation that a neuron receives and specific dendritic membrane properties define its computational capabilities. Until recently, there was relatively little known about the cellular and molecular mechanisms of dendritic growth, perhaps because dendrites were historically considered to be intrinsically determined, passive elements in the formation of connections in the nervous system. In the last few years, however, overwhelming evidence has accumulated indicating that dendritic growth is remarkably dynamic and responsive to environmental signals, including guidance molecules and levels and patterns of activity. This manuscript reviews our current understanding of the cellular and molecular mechanisms of dendritic growth, the influence of activity in sculpting specific patterns of dendritic arbors, and a potential integral role for dendrites in activity-dependent development of circuits in the nervous system.


An improved method for the isolation of islets from the beef pancreas.
An improved method for the isolation of islets from the beef and pig pancreas is described. The procedure involves the use of strips of Velcro that retain the partially-digested collagen during the isolation of islets by the collagenase technique. The spiny portion of the Velcro is ideally suited to retain the collagen and yet permit the separation of islets from the pancreatic parenchyma. A remarkably high yield of islets has been obtained from the beef and pig pancreas using this procedure.


Yeast artificial chromosome (YAC) vector cloning of the MYCN amplified domain in neuroblastomas.
About 25% of human neuroblastomas have amplification of the MYCN proto-oncogene, and this feature is associated with advanced stages of disease and rapid tumor progression. Estimates of the size of the amplicon range from 300 to 3,000 kb, with MYCN at or near the center. It has been determined previously that there is considerable conservation of the amplified sequences among different neuroblastomas, and very few rearrangements have been found. We decided to clone the entire amplified domain in YACs to assess the size, structure and organization of the amplified sequences in neuroblastomas and to identify novel or junctional sequences at the ends of an amplicon in double minutes (dmins) or in the germline. A YAC library was constructed from the SMS-KAN cell line, which contains dmins and has about 150 copies of MYCN. About 5,000 clones from this library were screened with MYCN as well as other probes from the amplified domain. To date, 16 YACs have been identified, with a median size of 240 kb (range 50-460 kb). These YAC clones can be arranged in a contiguous, linear map of greater than or equal to 1 megabase. Our data suggest that most neuroblastomas with MYCN amplification also amplify a region that is a megabase or more in size, making this the largest amplified domain cloned to date. The very large size suggests that there are other genes near MYCN whose expression is important in mediating the aggressive phenotype associated with MYCN amplification, or, alternatively, that the nearest origin of replication may be a considerable distance from MYCN.


Hydrogen sulfide as an oxygen sensor in trout gill chemoreceptors.
O2 chemoreceptors elicit cardiorespiratory reflexes in all vertebrates, but consensus on O2-sensing signal transduction mechanism(s) is lacking. We recently proposed that hydrogen sulfide (H2S) metabolism is involved in O2 sensing in vascular smooth muscle. Here, we examined the possibility that H2S is an O2 sensor in trout chemoreceptors where the first pair of gills is a primary site of aquatic O2 sensing and the homolog of the mammalian carotid body. Intrabuccal injection of H2S in unanesthetized trout produced a dose-dependent bradycardia and increased ventilatory frequency and amplitude similar to the hypoxic response. Removal of the first, but not second, pair of gills significantly inhibited H2S-mediated bradycardia, consistent with the loss of aquatic chemoreceptors. mRNA for H2S-synthesizing enzymes, cystathionine beta-synthase and cystathionine gamma-lyase, was present in branchial tissue. Homogenized gills produced H2S enzymatically, and H2S production was inhibited by O2, whereas mitochondrial H2S consumption was O2 dependent. Ambient hypoxia did not affect plasma H2S in unanesthetized trout, but produced a PO2-dependent increase in a sulfide moiety suggestive of increased H2S production. In isolated zebrafish neuroepithelial cells, the putative chemoreceptive cells of fish, both hypoxia and H2S, produced a similar approximately 10-mV depolarization. These studies are consistent with H2S involvement in O2 sensing/signal transduction pathway(s) in chemoreceptive cells, as previously demonstrated in vascular smooth muscle. This novel mechanism, whereby H2S concentration ([H2S]) is governed by the balance between constitutive production and oxidation, tightly couples tissue [H2S] to PO2 and may provide an exquisitely sensitive, yet simple, O2 sensor in a variety of tissues.


Fisetin inhibits laryngeal carcinoma through regulation of AKT/NF-κB/mTOR and ERK1/2 signaling pathways.
Targeting cancer cells is crucial for improving the efficiency of laryngeal cancer treatment. However, the signaling pathway and therapeutic strategy, related to the tumor, still need further research. Dietary flavonoid fisetin (3,3',4',7-tetrahydroxyflavone) found in many fruits and vegetables has been shown in preclinical studies to inhibit cancer growth through regulating cell cycle, apoptosis, angiogenesis, invasion and metastasis without causing any toxicity to normal cells. PI3K/AKT and ERK1/2 have been known as essential signaling pathways to modulate cell proliferation, apoptosis as well as autophagy via mTOR, Caspase-3 and NF-κB signals. In our study, flow cytometry and western blot assays suggested that apoptosis was induced by fisetin administration, promoting Caspase-3 expressions by regulating PI3K/AKT/NF-κB. Additionally, fisetin suppressed TU212 cells proliferation, which was linked with ERK1/2 inactivation. Further, the activation of PI3K/AKT-regulated mTOR was inhibited by fisetin, leading to transcription suppression and proliferation inhibition of TU212 cells. In vivo studies also showed that the tumor volume and weight of nude mice were reduced for fisetin use with KI-67 decrease and LC3II increase in tumor tissue samples. Together, our data indicated that fisetin had a potential role in controlling human laryngeal cancer through inhibiting tumor cell proliferation, inducing apoptosis and autophagy regulated by ERK1/2 and AKT/NF-κB/mTOR signaling pathways, which might provide a therapeutic strategy for laryngeal cancer inhibition in future.


Volume determinations in computed tomography.
Modern computed tomographic (CT) scanners allow reasonably high-resolution cross-sectional visualization of most viscera and masses. To determine the accuracy of CT volume estimation, CT volumes of inanimate objects, cadaver kidneys and spleens, and in vivo balloons were performed. Regions of interest were outlined by a hand-operated cursor, and the computer program calculated the cross-sectional area in square millimeters and in pixels. A second computer program used Simpson's rule to calculate the volume from these multiple cross-sectional area. Calculated CT volumes were within +/-10% of directly measured volumes. Tumor masses being treated by chemotherapy were followed up clinically and by CT. Volume changes determined by CT are believed to be equal to and frequently more sensitive than clinical examination. From our experimental CT and clinical experiences, accuracy can be affected by (1) respiratory movement; (2) rapid changes in in vivo blood volume; (3) low CT number-gradient at the objects' periphery; (4) observer error in cursor tracing of the desired structure; and (5) mathematical errors inherent in Simpson's rule. We conclude that CT can estimate volumes of CT definable masses and can be useful in following tumor response to therapy.


Prevalence, classification, and etiology of pain in Parkinson's disease: association between Parkinson's disease and fibromyalgia or chronic widespread pain.
Parkinson's disease (PD) is characterized by resting tremor, slow and decreased movement (hypokinesia and akinesia), rigidity, postural instability, problems with gait, and coordination. The prevalence of PD is between 0.1% and 0.3% in the general population and between 1% and 2% in persons 65 years of age or older. Patients with PD are more likely to suffer from pain. Indeed, the chief complaint of patients with severe motor disturbance and severe pain is pain rather than motor disturbance. Fibromyalgia (FM) is defined by widespread pain (pain in the left and right sides of the body, pain above the waist, pain below the waist, and axial skeletal pain) for more than 3 months and the presence of at least 11 of the 18 specified tender points. FM and chronic widespread pain (CWP), which is usually an incomplete form of FM, cause pain in the musculoskeletal region, but their etiologies are unknown. Therefore, it is almost impossible to determine whether or not pain in the musculoskeletal region is in the musculoskeletal origin. We suspect that dysfunction or degeneration of the nerves that control pain, mind, and movement in the brain causes FM, depression, and PD, respectively. When pain in PD is discussed, FM and CWP should be considered because their prevalence is high. Patients with PD may be likely to suffer from FM and CWP; however, the prevalence of FM and CWP in patients with PD has not been reported. Here, we discuss the relationship between PD and FM or CWP.


Criterion for material selection in design of bulk piezoelectric energy harvesters.
Vibration energy harvesting has gained tremendous attention in the past decade and continues to grow rapidly. There are various transduction mechanisms for converting the vibration energy into electrical energy, out of which the piezoelectric mechanism has been shown to provide advantages at the micro-to-meso scale. In the past few years, several studies have tried to address the question of which piezoelectric composition is better for energy harvesting; however, discussion on this subject continues. The intent of this letter is to provide an answer for this question through a simple criterion which can be used in routine material evaluation.


HIV infection and AIDS in Jerusalem: a microcosm of illness in Israel.
During the decade 1985-94, 158 individuals (124 males, 34 females) with HIV/AIDS were seen at the AIDS clinic of Hadassah University Hospital. They comprised 10% of the total reported number of HIV-infected Israelis and included 6% of all reported HIV-infected Ethiopians and all HIV-infected West Bank Palestinians. Most individuals (82%) were self-referred; 12% were detected through the local HIV screening clinic. Risk groups for AIDS were homo/bisexuality (39%), heterosexual relations (30%), intravenous drug abuse (15%), contaminated blood transfusions (9%), born to HIV+ mothers (1%), and hemophilia (< 1%). In 5% (all males) no risk factor was given. Of the 158 individuals 29 were Ethiopian immigrants, 25 West Bank Palestinians, and the rest were local Israelis and tourists. Of the 34 females seen, 21 acquired HIV heterosexually, 6 were intravenous drug abusers, 5 were recipients of contaminated transfusions, and 2 were infants. An initial clinical presentation with full-blown AIDS was observed in 30% of the individuals. The range of clinical presentations was similar to that seen in western countries, with Pneumocystis carinii pneumonia, cytomegalovirus retinitis, mycobacterial infections and Kaposi's sarcoma seen most frequently. One case of disseminated cutaneous leishmaniasis was observed. The mortality rate during the follow-up was 55%. In 35% of individuals the immediate cause of death was severe bacterial infections. The Palestinian subpopulation presented at an advanced stage of the disease with a high incidence of transfusion recipients, while most HIV-infected Ethiopians presented with asymptomatic disease transmitted heterosexually. HIV/AIDS as seen at Hadassah University Hospital during 1985-94 exhibited the mixed form of Euro-American AIDS with additional facets of recently introduced African infection.


Analysis of lipoprotein lipase haplotypes reveals associations not apparent from analysis of the constituent loci.
Simultaneously analysing genotype effects at several closely-linked loci may be preferable to analysing them separately, but can be difficult, due to multiple genotype classes, small class sizes, and non-independence induced by associations among loci. Analysis of haplotype effects offers an alternative approach. We studied effects of haplotypes comprising 3 loci (5' to 3': PvuII, HindIII, and Ser 447 -Stop) in the lipoprotein lipase (LPL) gene on plasma lipid levels and LPL activity, in 807 Dutch males with coronary atherosclerosis. We analysed haplotype effects in individuals for whom haplotypes could either be determined unequivocally or inferred with high probability, using contrasts suggested by likely evolutionary relationships among the haplotypes. One haplotype was associated with significantly higher total cholesterol, while another was associated with significantly lower triglyceride levels. Though these two haplotypes had generally opposite effects on lipids, both were associated with significantly higher LPL activity. In genotype analyses, the HindIII (-) allele was associated with higher LPL activity; however, one haplotype bearing it had no significant effect on LPL activity. Haplotypes thus provided more information than genotypes alone would have. The two haplotypes with consistently different effects on lipid levels despite similar effects on LPL activity, provide further evidence that aspects of LPL biology, apart from its catalytic function in lipolysis, may mediate its effects on plasma lipids at least in coronary artery disease patients.