Evaluating a Cumaean Sibyl: Domenichino or later? A multi analytical approach.
The purpose of this study is to clarify important details about a Cumaean Sibyl painting that is preserved in a private collection. This work, bearing neither signature nor date, has never undergone conservation. It was executed after Domenichino's Cumaean Sibyl, a work known to have been used as a model by many 18th century painters. Investigation of the anonymous artist's painting technique and identification of its constituent materials were facilitated by employing optical microscopy (OM), scanning electron microscopy (SEM/EDS), FTIR and microRaman spectroscopies and high performance liquid chromatography with diode array detection (HPLC-DAD). The painter's palette comprised lead white, yellow ochre, lead-antimonate yellow (Naples yellow), cinnabar, cochineal lake, madder lake, haematite, Prussian blue and carbon black. The detection of Prussian blue (synthesized in 1704 and widely used as artist's pigment after 1750) was decisive in establishing the work's authenticity for, as such, it cannot be attributed either to Domenico Zampieri (1581-1641) or to his apprentices. In addition, the identification of Naples yellow, which prevailed in the period from 1750 to 1850, supports this statement. Nevertheless, its elaborate painting technique strongly suggests an artist greatly influenced by the Renaissance masters. A comparison of its stylistic features with those of the Cumaean Sibyl of Angelica Kauffmann (1741-1807), a prolific 18th century artist known to have studied and to have copied Domenichino's Sibyl, reveals significant similarities between the two in composition and palette. The unsigned Sibyl, therefore, could well be by Kauffmann.


Studying Autophagy Using a TMT-Based Quantitative Proteomics Approach.
Maintenance of cellular homeostasis through regulated degradation of proteins and organelles is a defining feature of autophagy. This process itself is tightly regulated in a series of well-defined biochemical reactions governed largely by the highly conserved ATG protein family. Given its crucial role in regulating protein levels under both basal and stress conditions such as starvation and infection, genetic or pharmacological perturbation of autophagy results in massive changes in the cellular proteome and impacts nearly every biological process. Therefore, studying autophagy perturbations at a global scale assumes prime importance. In recent years, quantitative mass spectrometry (MS)-based proteomics has emerged as a powerful approach to explore biological processes through global proteome quantification analysis. Tandem mass tag (TMT)-based MS proteomics is one such robust quantitative technique that can examine relative protein abundances in multiple samples (parallel multiplexing). Investigating autophagy through TMT-based MS approach can give great insights into autophagy-regulated biological processes, protein-protein interaction networks, spatiotemporal protein dynamics, and identification of new autophagy substrates. This chapter provides a detailed protocol for studying the impact of a dysfunctional autophagy pathway on the cellular proteome and pathways in a healthy vs. disease (virus infection) condition using a 16-plex TMT-based quantitative proteomics approach. We also provide a pipeline on data processing and analysis using available web-based tools.


Zeaxanthin is bioavailable from genetically modified zeaxanthin-rich potatoes.
The carotenoid zeaxanthin accumulates in the human macula lutea and protects retinal cells from blue light damage. However, zeaxanthin intake from food sources is low. Increasing zeaxanthin in common foods such as potatoes by traditional plant breeding or by genetic engineering could contribute to an increased intake of this carotenoid and, consequently, to a decreased risk of age-related macular degeneration. Our aim was to investigate whether zeaxanthin from genetically modified zeaxanthin-rich potatoes is bioavailable in humans. Three men participated in this randomized, controlled double-blinded, crossover pilot study. All subjects consumed 1,100 g of mashed potatoes, either genetically modified (Solanum tuberosum L. var. Baltica GM47/18; 3 mg zeaxanthin) or wild-type control potatoes (Solanum tuberosum L. var. Baltica; 0.14 mg zeaxanthin). A second treatment was followed after a 7-day wash-out period. The concentration of zeaxanthin was significantly increased in chylomicrons after consumption of genetically modified potatoes and 0.27 mg of the 3 mg zeaxanthin dose could be detected in chylomicrons. Consumption of control potatoes had no effect on concentrations of zeaxanthin in chylomicrons. After normalization of chylomicron zeaxanthin for plasma triacylglycerol, the time course of zeaxanthin concentrations peaked at 7 h after consumption of genetically modified potatoes. There were no significant differences in the concentrations of other major potato carotenoids such as lutein and beta-carotene in chylomicrons after consumption of genetically modified and wild type control potatoes. Thus, consumption of zeaxanthin-rich potatoes significantly increases chylomicron zeaxanthin concentrations suggesting that potentially such potatoes could be used as an important dietary source of zeaxanthin.


The effect of benzoyl peroxide on cutaneous micro-organisms in vitro.
The survival curves of cutaneous micro-organisms in the presence of benzoyl peroxide were investigated. All the curves exhibited a shoulder prior to exponential cell death. Benzoyl peroxide was lethal to the cutaneous organisms tested and they varied in sensitivity increasing as follows: Propionibacterium acnes, Staphylococcus capitis, Staph. epidermidis, Staph. hominis, Prop. avidum, Prop. granulosum and Pityrosporum ovale.


The First Brazilian Cardiovascular Registry of Atrial Fibrillation: Primary Results of the RECALL Study.
It is estimated that atrial fibrillation (AF) affects approximately 1.5 million people in Brazil; however, epidemiological data are limited. We sought to evaluate the characteristics, treatment patterns, and clinical outcomes in patients with AF in Brazil by creating the first nationwide prospective registry. RECALL was a multicenter, prospective registry that included and followed for 1 year 4,585 patients with AF at 89 sites across Brazil from April 2012 to August 2019. Patient characteristics, concomitant medication use, and clinical outcomes were analyzed using descriptive statistics and multivariable models. Of 4,585 patients enrolled, the median age was 70 (61, 78) years, 46% were women, and 53.8% had permanent AF. Only 4.4% of patients had a history of previous AF ablation and 25.2% had a previous cardioversion. The mean (SD) CHA2DS2-VASc score was 3.2 (1.6); median HAS-BLED score was 2 (2, 3). At baseline, 22% were not on anticoagulants. Of those taking anticoagulants, 62.6% were taking vitamin K antagonists and 37.4% were taking direct oral anticoagulants. The primary reasons for not using an oral anticoagulant were physician judgment (24.6%) and difficulty in controlling (14.7%) or performing (9.9%) INR. Mean (SD) TTR for the study period was 49.5% (27.5). During follow-up, the use of anticoagulants and INR in the therapeutic range increased to 87.1% and 59.1%, respectively. The rates/100 patient-years of death, hospitalization due to AF, AF ablation, cardioversion, stroke, systemic embolism, and major bleeding were 5.76 (5.12-6.47), 15.8 (14.6-17.0), 5.0 (4.4-5.7), 1.8 (1.4-2.2), 2.77 (2.32-3.32), 1.01 (0.75-1.36), and 2.21 (1.81-2.70). Older age, permanent AF, New York Heart Association class III/IV, chronic kidney disease, peripheral arterial disease, stroke, chronic obstructive pulmonary disease, and dementia were independently associated with increased mortality while the use of anticoagulant was associated with lower risk of death. RECALL represents the largest prospective registry of patients with AF in Latin America. Our findings highlight important gaps in treatment, which can inform clinical practice and guide future interventions to improve the care of these patients.


Somato-vesical reflexes in chronic spinal cats.
The effects of afferent volleys in hindlimb cutaneous and muscle nerves on vesical tone and contractility and on the discharges in pelvic nerves to the bladder were measured in anesthetized CNS-intact and 2-19 months chronic spinal cats. In chronic spinal cats volleys in group III and IV fibers increased the tone of the quiet, empty bladder (excitatory somato-vesical reflex). The same volleys inhibited the slow, large, rhythmic micturition contractions of the expanded bladder (inhibitory somato-vesical reflex). In CNS intact cats single or short tetanic volleys induced a reflex discharge in pelvic vesical nerve branches with 3 distinct components. These reflexes could be observed during micturition contractions, not markedly between the contractions or when the bladder was empty and quiet. The latencies of the 3 components were 90, 320 and 770 ms, respectively. The two early components (AI- and A2-reflex) were evoked by volleys in group II and III hindlimb afferents. The late component (C-reflex) was induced by group IV volleys. In chronic spinal cats a group II and III-induced A-reflex (latency 90 ms) and a group IV-induced C-reflex (latency 340 ms) were observed. The central pathways and the physiological significance of the various somato-vesical reflexes are discussed.


Complex Approach to Xenobiotics Hepatotoxicity Testing using a Microfluidic System.
We analyzed hepatotoxicity of three drugs: acetaminophen, metformin, and isoniazid. Spheroids of differentiated HepaRG cells cultured under microfluidic conditions were used as the model. Acute toxicity of substances was assessed by analyzing cell viability, while lactate concentration in the culture medium was used as the potential marker for evaluation of chronic exposure and non-lethal side effects of xenobiotics. The results were compared with mitochondrial activity and DNA fragmentation data. The efficiency and possibility of applying the integrated approach for assessment of drug hepatotoxicity are discussed.


"Superinduction" of tyrosine aminotransferase by actinomycin D: a reevaluation.
Reexamination of the effects of actinomycin D (AMD) on the intracellular level and rate of synthesis of tyrosine aminotransferase (TAT) in hepatoma tissue culture (HTC) cells reveals that much apparent controversy can be resolved with acknowledgment of the multi-faceted nature of this inhibitor's action. AMD can slow overall protein synthesis and inhibit the degradation of both TAT and its mRNA as well as block the synthesis of RNA. The extent of these secondary actions of the inhibitor depend somewhat upon the growth condition of the cells. The effects of cordycepin (3'-deoxyadenosine) on the metabolism of TAT and its mRNA are also complex, but differ in several respects from those of AMD.


Differential Inhibition of Helicoverpa armigera (Hubner) Gut Proteinases by Proteinase Inhibitors of Okra and It's Wild Relatives.
The seeds of ten genotypes and twenty-nine wild relatives of okra were analysed for the presence of trypsin, chymotrypsin, and Helicoverpa gut proteinases (HGPs) inhibitors (HGPIs), with the aim to identify potent inhibitors of H. armigera gut proteinases. Proteinase inhibitors (PIs) obtained from wild relatives of okra exhibited stronger inhibition of HGPs than the genotypes of okra. In in vitro inhibitory assay against HGPs, A. tuberculatus 90396 and 90515 showed high tryptic inhibitory (71.8% and 69.2%), chymotryptic inhibitory (68.5% and 66.2%), and Helicoverpa gut proteinase activity (70.2% and 68.2%). In electrophoretic profile showed the same variation in the number of trypsin inhibitors (TIs), chymotrypsin Inhibitors (CIs), and HGPIs isoforms with different intensities, whereas genotypes of okra mostly showed monomorphic profile. Maximum eight HGPIs isoforms were found in A. tuberculatus (90396 and 90515). In bioassay studies, significant reduction in weight of H. armigera larvae was found, when larvae fed on PIs obtained from A. tuberculatus (90396 and 90515). Thus, the result of the present investigation indicates that further exploration of PIs obtained from A. tuberculatus (90396 and 90515) will be helpful for developing PIs-based insect resistance management strategies.


Impact of surgeon volume on patient safety in laparoscopic gynecologic surgery.
To determine the impact of surgeon volume on patient outcomes in gynecologic laparoscopic surgery. We reviewed all patients who underwent a laparoscopic procedure between January 2000 and December 2008. Surgeons were divided into 3 groups based on surgical volume. The mean number of surgeries per year was calculated for each surgeon. Means were categorized into 3 groups. The low-, medium-, and high-volume surgeon groups were compared with respect to level of surgical complexity and intraoperative and postoperative complications. The study included 829 surgeries. Low-volume surgeons (n=5) performed 5 (31.3%) low-complexity, 10 (62.5%) intermediate-complexity, and 1 (6.3%) high-complexity procedures. Medium-volume surgeons (n=6) performed 26 (11.1%) low-complexity, 203 (86.8%) intermediate-complexity, and 5 (2.1%) high-complexity procedures. High-volume surgeons (n=5) performed 47 (8.1%) low-complexity, 439 (75.8%) intermediate-complexity, and 93 (16.1%) high-complexity procedures. The distribution of surgical complexity was significantly different between the 3 groups of surgeons defined by volume (P<0.001). Conversion rates were higher for low-volume surgeons when compared to high-volume surgeons (18.8% vs. 5.2%; P=0.04). Similarly, overall complication rates (<30 days) were higher for low-volume surgeons compared to high-volume surgeons (31.3% vs. 17%, P=0.003). Mean length of hospital stay was longer for low-volume (2.4 days) than for medium-volume (1.3 days) and high-volume surgeons (1.6 days) (P=0.003). High- and medium-volume gynecologic laparoscopists performed a greater proportion of intermediate- and high-complexity procedures than did low-volume surgeons. High-volume surgeons have a lower rate of conversions, overall postoperative complications, and shorter mean length of hospital stay when compared to low volume surgeons.


A case of dense pigment deposition of the posterior lens capsule.
Pigment dispersion syndrome (PDS) is a well-known entity which can lead to pigmentary glaucoma (PG). This case report presents a rare presentation of PG with bilateral dense pigment deposits of the posterior lens capsule. A 72-year-old male came for his first appointment due to an asymmetric worsening of visual acuity. The examination showed unilaterally severely increased intraocular pressure, bilateral dense pigment deposition of the posterior lens capsule, and a shallow unilateral optic disk excavation. Gonioscopy revealed moderate pigmentation of the angle and a concave configuration of the peripheral iris in both eyes. The standard slit lamp examination showed no transillumination defects of either iris. Optical coherence tomography showed retinal nerve fiber layer (RNFL) thinning in the peripapillary and macular regions. An antiglaucoma medication was prescribed with a good lowering effect. Pigment deposition of the posterior lens capsule, which has been rarely reported, is a possible important sign of PDS or PG.


Glycoconjugates in exfoliation syndrome. A lectin histochemical study of the ciliary body and lens.
The binding of ten biotin-coupled lectins with different carbohydrate specificities to exfoliative material and neighbouring tissues was studied in 16 formalin-fixed and paraffin-embedded human eyes. Eight of the eyes were exfoliation positive while the rest were exfoliation negative. Exfoliative material reacted intensely with Lens culinaris (LCA), Canavalia ensiformis (ConA) and Ricinus communis (RCA-I) agglutinins. Positive reaction was also seen with wheat germ (WGA), peanut (PNA) and soybean (SBA) agglutinins. The superficial zonular lamella, zonular fibers and the non-pigmented epithelium of the ciliary body had a rather similar lectin-binding profile to exfoliative material. The lens capsule was essentially unreactive with all the lectins used. The lens epithelium reacted faintly with ConA, LCA, WGA and RCA-I. Pre-treatment with neuraminidase to remove sialic acid resulted in increased binding of PNA and SBA to exfoliative material, zonular fibers and the zonular lamella, and in decreased binding of WGA to the non-pigmented epithelium of the ciliary body, zonular fibers and the zonular lamella. The results indicate that alpha-mannosyl, beta-galactosyl, N-acetyl-D-glucosaminyl and N-acetylneuraminic acid residues are present in glycoconjugates of exfoliative material and neighbouring tissues.


Facile synthesis of nanostructured TiNb2O7 anode materials with superior performance for high-rate lithium ion batteries.
One-dimensional nanostructured TiNb2O7 was prepared by a simple solution-based process and subsequent thermal annealing. The obtained anode materials exhibited excellent electrochemical performance with superior reversible capacity, rate capability and cyclic stability.


Telemetric recording of neuronal activity.
A telemetric system is described which allows the wireless registration of extracellular neuronal activity and vocalization-associated skull vibrations in freely moving, socially living squirrel monkeys (Saimiri sciureus). The system consists of a carrier platform with numerous guiding tubes implanted on the skull. Custom-made microdrives are mounted on the platform, allowing the exploration of two electrode tracks at the same time. Commercially available quartz-insulated platinum-tungsten microelectrodes are used. The electrodes can be moved over a distance of 8-10 mm by turning a screw on the microdrive. Vocalization-associated skull vibrations are recorded with a piezo-ceramic element. Skull vibration signal and the signals from the two microelectrodes are fed into separate transmitters having different carrier frequencies. The signals are picked up by an antenna in the animal cage and are sent to three receivers in the central laboratory. Here, the signals are transferred via an analog/digital interface to a personal computer for data analysis and to a video recorder for long-term storage. The total weight of the head mount including carrier platform, microdrive, electrodes, skull vibration sensor, three transmitters, and protection cap is 32 g. The transmitters are powered with two rechargeable lithium batteries, allowing about 8 h of continuous recording. Reliable signal transmission is obtained over a distance of about 2 m. Recording stability allows to follow the activity of specific neurons up to several hours, with no movement artefacts during locomotion.


Effects of metronidazole on mesophilic and thermophilic fermentation: Biodegradation mechanisms, microbial communities, and reversibility.
Metronidazole (MNZ), an antibiotic that is specifically used for the treatment of anaerobic infections, may inhibit anaerobic fermentation. This work was designed to understand the fate and effects of MNZ in mesophilic fermentation (MF) and thermophilic fermentation (TF), respectively. The results showed that the removal of MNZ mainly occurred via biodegradation, rather than adsorption, and that MNZ could be completely degraded by opening the imidazole ring. MFs were more strongly inhibited by MNZ than TFs. MNZ concentration increased from 0 to 25 mg/L, hydrogen yield (HY) decreased from 167.5 to 16.8 mL/g glucose (90.0% decrease), and butyrate yield almost completely disappeared in MFs, whereas in TFs, HY decreased only from 101.1 to 89.3 mL/g glucose (11.7% decrease), and ethanol yield increased by 39.8%. Illumina MiSeq sequencing analysis showed that MNZ reduced the abundance of hydrogen-producing bacteria. Furthermore, the inhibition of MNZ on anaerobic fermentation was reversible.


Store-operated Ca2+ channels blockers inhibit lipopolysaccharide induced astrocyte activation.
The destruction of calcium homeostasis is an important factor leading to neurological diseases. Store-operated Ca(2+) (SOC) channels are essential for Ca(2+) homeostasis in many cell types. However, whether SOC channels are involved in astrocyte activation induced by lipopolysaccharide (LPS) still remains unknown. In this study, we used LPS as an exogenous stimulation to investigate the role of SOC channels in astrocyte activation. Using calcium imaging technology, we first found that SOC channels blockers, 1-[h-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF-96365) and 2-aminoethyldiphenyl borate (2-APB), inhibited LPS induced [Ca(2+)]i increase, which prompted us to speculate that SOC channels may be involved in LPS induced astrocyte activation. Further experiments confirmed our speculation shown as SOC channels blockers inhibited LPS induced astrocyte activation characterized as cell proliferation by MTS and BrdU assay, raise in glial fibrillary acidic protein expression by immunofluorescence and Western Blot and secretion of interleukin 6 (IL-6) and interleukin 1β (IL-1β) by ELISA. So, our studies showed that SOC channels are involved in LPS-induced astrocyte activation.


Graft derived reafferentation of host spinal cord is not necessary for amelioration of lesion-induced deficits: possible role of migrating grafted astrocytes.
The present study explores the ability of fetal spinal cord homografts into lesioned host C3 fasiculus gracilis to influence the expected deterioration of hindlimb performance following this lesion. Rats were trained to traverse a narrow platform for a water reward. Animals were ranked for hindlimb performance utilizing slips, recovery and manner of traversing the platform. After training the animals, numbers were recorded, laminectomy performed at C3 and subject fasiculus gracilis (FG) bilaterally aspirated. Half the subjects were randomly selected for implantation of two, one mm segments of 14 day gestation cervical spinal cord. Recorded lesion-only and lesion-transplanted animals were tested 21, 30, 45, 60 and 90 days later. C3 fetal transplants significantly decreased the severity of hindlimb deficit at 21 and 90 days postlesion (p less than 0.05). The C1-FG of both groups contained no nerve fibers. However, the host nucleus gracilis of lesion-transplant animals contained normal sizes and numbers of neurons whereas the lesion-only group did not. This neuronal maintenance may have been due to factor(s) secreted by transplant derived astrocytes which migrated at 0.72-0.76 mm/day and reside in the host nucleus gracilis.


Development of a versatile reporter assay for studies of retinol uptake and metabolism in vivo.
The two isomers of retinoic acid (RA), all-trans RA and 9-cis RA, are produced in several tissues in order to allow specific control of target gene transcription. Given the high potency of these receptor ligands, it seems likely that the cellular uptake and metabolic activation of the precursor, retinol (vitamin A), should be a highly regulated process. Several retinol dehydrogenases and components involved in the downstream events have been identified and partially characterized. However, less is known about the cellular uptake of retinol, and the isomerase activity giving rise to the 9-cis and 11-cis branches of the pathway. In this work, we show that the 9-cis RA biosynthesis pathway can be fully reconstituted in cultured HEK293A cells expressing a reporter system, including an endogenous isomerase activity converting all-trans retinol into 9-cis retinol. This assay allows for functional studies of known components, as well as screening for yet unidentified genes involved in the pathway. In addition to free all-trans retinol, we find that these cells can take up retinol from plasma retinol binding protein (RBP) by a mechanism that can be efficiently inhibited by blocking antibodies, suggesting that the uptake may involve a cellular receptor. We also demonstrate that overexpression of CRBPI can drive the accumulation of intracellular retinol from unbound retinol added to the medium. Thus, this versatile cellular assay can be used to study several aspects of retinol uptake and metabolism in vivo.


Electrospun nanofiber based on Ethyl cellulose/Soy protein isolated integrated with bitter orange peel extract for antimicrobial and antioxidant active food packaging.
The present work was aimed to produce a novel bioactive nanofiber (NFs) based on Ethyl cellulose (EC), Soy protein isolated (SPI), and containing Bitter orange peel extract (BOPE) by electrospinning technology. The EC/SPI NFs were formulated with different weight ratios of 1:1, 2:1, and 1:2 denoted as ES11, ES21, and ES12, respectively, and investigated by several analyses. Based on the obtained results, the maximum hydrogen interactions between these two polymers, ES11 NFs offered a uniform morphology without bead with the diameter of 185.33 nm as a result of the compatibility of the polymer solutions of EC and SPI. Moreover, appropriate thermal stability was presented along with more porosity (78%), maximum water vapor transmission rate (657 g/m2.24h), good tensile stress (6.12 MPa), and acceptable water contact angel (82.3°). Therefore, ES11 NFs were selected as the optimal sample for incorporation of the BOPE as the antibacterial and antioxidant agent. According to the antioxidant activity test, the highest concentration (20% wt) of this extract increased the antioxidant activity of NF around 64.7% and also inhibited the growth of pathogenic bacteria (S. areus, and E. coli). Therefore, the ES11 electrospun NFs containing 20% BOPE can be a beneficial system to increase the safety and quality of foods.


Unrelated cord blood transplantation vs matched unrelated donor bone marrow transplantation: the risks and benefits of each choice.
Matched sibling BMT is the treatment of choice for many malignant and non-malignant diseases. Unfortunately, donor availability limits its applicability with only approximately one-third of patients having HLA-matched family members. For the remaining two-thirds of patients who need an allogeneic procedure, an alternative donor is required. Matched unrelated donor (MUD) bone marrow transplants have proven useful for many of these patients, however, matched donors may be unavailable or too time-consuming to find. In addition, MUD transplants are associated with high morbidity and mortality secondary to severe graft-versus-host disease (GVHD), organ toxicity, and infections. For those patients without a matched related or unrelated volunteer donor, unrelated umbilical cord blood transplantation (UCBT) is a feasible alternative. UCBT has been associated with a lower incidence and severity of GVHD than MUD BMT, allowing successful transplantation in the HLA-mismatched setting, thus increasing the donor pool. This manuscript reviews the available literature regarding these two procedures, assesses their risk/benefit ratio, and arrives at several conclusions regarding their relative indications.


Bioactive glass-based composites for the production of dense sintered bodies and porous scaffolds.
Recently several attempts have been made to combine calcium phosphates, such as β-tricalcium phosphate (β-TCP) and, most of all, hydroxyapatite (HA), with bioactive glasses of different composition, in order to develop composites with improved biological and mechanical performance. Unfortunately, the production of such systems usually implies a high-temperature treatment (up to 1300 °C), which may result in several drawbacks, including crystallization of the original glass, decomposition of the calcium phosphate phase and/or reactions between the constituent phases, with non-trivial consequences in terms of microstructure, bioactivity and mechanical properties of the final samples. In the present contribution, novel binary composites have been obtained by sintering a bioactive glass, characterized by a low tendency to crystallize, with the addition of HA or β-TCP as the second phase. In particular, the composites have been treated at a relatively low temperature (818 °C and 830 °C, depending on the sample), thus preserving the amorphous structure of the glass and minimizing the interaction between the constituent phases. The effects of the glass composition, calcium phosphate nature and processing conditions on the composite microstructure, mechanical properties and in vitro bioactivity have been systematically discussed. To conclude, a feasibility study to obtain scaffolds for bone tissue regeneration has been proposed.


Immunological analogy between allograft rejection, recurrent abortion and pre-eclampsia - the same basic mechanism?
There are still controversies concerning the role of immunological mechanisms engaged both in recurrent abortions (RA) and pre-eclampsia (PE). According to some opinions, recurrent miscarriage is comparable to organ-specific autoimmune disease. Analysis of immune reactions shows that graft rejection shares many similar mechanisms with RA and PE. This fact allows us to conclude that rejection of transplanted alloantigenic organs and pregnancy loss have probably the same evolutionary origin. Subsets and functions of immunocompetent cells (T CD4, suppressor gammadeltaT, cytotoxic T CD8, Treg, Tr1, uterine NK cells), over-activation of innate immunity (activation of NK cytotoxic cells, macrophages, neutrophils and complement), changes of Th1/Th2 cytokine balance (IL-2, IL-12, IL-15, IL-18, IFNgamma, TNFalpha vs. IL-4, IL-10, TGFbeta), importance of HLA-G molecule, CD200/CD200R interaction, over-expression of adhesion molecules, fgl2 prothrombinase activation and stimulation of IDO and HO expression, all suggest that RA and PE are syndromes of fetal allograft rejection, and not organ-specific autoimmune diseases. According to that supposition, an analogy might exist between acute graft rejection and recurrent abortion, and between chronic graft rejection and pre-eclampsia.


The role of nutrient presence on the adhesion kinetics of Burkholderia cepacia G4g and ENV435g.
The adhesion kinetics of Burkholderia cepacia G4g and ENV435g have been investigated in a radial stagnation point flow (RSPF) system under well-controlled hydrodynamics and solution chemistry. The sensitivity of adhesion behavior to nutrient condition was also examined. Supplementary cell characterization techniques were conducted to evaluate the viability, hydrophobicity, electrophoretic mobility, size, and charge density of cells grown in both nutrient rich Luria broth (LB) and nutrient poor basal salts medium (BSM). Comparable adhesion kinetics were observed for the wild-type (G4g) and mutant (ENV435g) grown in the same medium; however, the attachment efficiency increased with the level of nutrient presence for both cell types by approximately 60%. Nutrient condition altered deposition due to its impact on the surface charge characteristics and size of the cells. Adhesion behavior was consistent with expectations based on classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory for colloidal interactions, as the adhesion efficiency increased with ionic strength. However, the results also suggest the involvement of non-DLVO type interactions that influence cell adhesion. Systematic experimentation with B. cepacia in the RSPF system demonstrated that the ENV435g mutant is not "adhesion deficient"; rather, adhesion for both the G4g and ENV435g was a function of the nutrient condition and resulting cell surface chemistry.


1H NMR-based metabolomics investigation on the biological effects of paralytic shellfish toxin in mussel Mytilus trossulus.
Paralytic shellfish poisoning is a global issue that would benefit from additional screening methods and rapid testing capacities. In this study, we applied 1H NMR spectroscopy-based metabolomics to identify biomarkers of Paralytic Shellfish Toxin (PST) exposure. We characterized the metabolic phenotypes of field-collected Alaskan mussels Mytilus trossulus across a wide range of bioaccumulated PST levels, from 0 to 1590 μg/100 g. A between-level grouping emerged for high (740-1590 μg/100 g) compared to low/non-detect (0-3.91 μg/100 g) PST levels. High levels of PST contamination in mussels were consistent with alterations to energy and amino acid metabolism, and disturbances in osmoregulation. This research demonstrates the effectiveness of 1H NMR-based metabolomics in elucidating the biological effects of paralytic shellfish toxin on the health of wild mussel populations, spatial variation, and identifies a metabolic signature indicative of PST contamination in Mytilus trossulus for potential use in a PSP biomarker panel.


Natural deep eutectic solvents protect RNA from thermal-induced degradation.
RNA is a fundamental nucleic acid for life and it plays important roles in the regulation of gene transcription, post-transcriptional regulation, and epigenetic regulation. Recently, the focus on this nucleic acid has significantly increased due to the development of mRNA vaccines and RNA-based gene therapy protocols. Unfortunately, RNA based products show constrains mainly owing to instability and easy degradability of the RNA molecules. Indeed, unlike the DNA molecule which has a great intrinsic stability, RNA is more prone to degradation and this process is accelerated under thermal treatment. Here we describe a method that involves the use of Natural Deep Eutectic Solvents (NaDES) capable of slowing down RNA degradation process. Our results show that this technology seems suitable for improving the stability of specific RNA molecules particularly susceptible to thermal-induced degradation. Therefore, this technique represents a valuable tool to stabilize RNA molecules used in gene therapy and mRNA vaccines.


Stress in the city? Coyote hair cortisol varies with intrinsic and extrinsic factors within a heavily urbanized landscape.
Wildlife living in proximity to people are exposed to both natural and anthropogenic factors that may influence cortisol production associated with stress response. While some species, including coyotes (Canis latrans), have become commonplace in developed areas throughout North America, urban individuals still must navigate ever-changing, novel environments and cope with frequent disturbance. Given that coyotes are relatively large predators compared to most other urban wildlife, they face unique pressures such as crossing roadways to use suitable habitat fragments and are at a greater risk of being detected and experiencing negative human interactions. To assess whether urbanization influences hypothalamic-pituitary-adrenal axis activity in free-ranging coyotes, we analyzed cortisol concentration in hair samples from 97 coyotes residing across the urbanization gradient within the Greater Chicago Metropolitan area. As the proportion of developed landcover within coyote home ranges increased, coyotes experienced more stress. Body condition and social status also had strong relationships with stress. Animals in poorer body condition experienced more stress and subordinate coyotes experienced less stress than alphas. We also found some evidence that stress varied seasonally and among different age classes. Understanding how intrinsic and extrinsic factors influence endocrine activity in urban carnivores is vital for predicting how hormone production and related behavioral patterns may change in future populations as more areas become developed.


On a therapeutic target point for the non-operative treatment of intestinal obstruction.
The period necessary to obtain a 95% improvement level through non-operative treatment of simple obstruction of the small bowel was found to be about six days, based on a study of 29 patients selected from approximately 1,000 clinical cases of verified or suspected intestinal obstruction. An additional experimental study confirmed the case-screening criteria.


Vasoactive intestinal and calcitonin gene-related peptides, tyrosine hydroxylase and nitrergic markers in the innervation of the rat central retinal artery.
The vascular supply of the optic nerve has been studied with different methods including corrosion casts both in humans and in other mammals. In man, primates and some other mammals, such as the rat, a distinct central retinal artery accompanies the optic nerve, and runs through the lamina cribosa to reach the optic nerve head. Similarities between human and rat central retinal artery could serve to understand changes in the autonomic perivascular innervation in glaucoma using the rat as an animal model. Nitric oxide, calcitonin gene-related peptide, neuropeptide Y, substance P and vasoactive intestinal peptide have been identified around the monkey central retina artery. Innervation of the rat central artery, however, has not been described in detail. Using immuno- and histochemical methods, the present study investigates the peptidergic, adrenergic and nitrergic innervation of the rat posterior ciliary artery as well as the central retina artery. Numerous nitric oxide positive nerve fibers were visualized posterior and anterior to the lamina cribosa of the optic nerve. They colocalized with NADPH-diaphorase positive fibers, which could also be observed in two of six specimens studied at the level of the optic nerve head. Calcitonin gene-related peptide, tyrosine hydroxylase, and VIP positive fibers were also observed surrounding the vessels of the rat optic nerve. The presence of neuronal nitric oxide/NADPH-diaphorase and vasoactive intestinal peptide positive nerve fibers surrounding the posterior ciliary and central retinal arteries indicates a vasodilator effect in the rat optic nerve. Tyrosine hydroxylase positive innervation indicates the presence of sympathetic activity, and calcitonin gene-related peptide positive fibers indicate sensory innervation by trigeminal primary efferents.


Continuous versus cyclic oral contraceptives after laparoscopic excision of ovarian endometriomas: a systematic review and metaanalysis.
In the lack of evidence consistently supporting the use of continuous vs cyclic oral contraceptives after surgery for endometriosis, we conducted a systematic review and metaanalysis with the objective of comparing a continuous vs a cyclic oral contraceptive schedule administered after surgical excision of ovarian endometriomas. A PubMed, MedLine, and Embase search through December 2014 was conducted, with the use of a combination of key words and text words related to endometrioma, endometriosis, oral contraceptives, oral estroprogestins, laparoscopy, and surgery. Studies directly comparing a continuous vs a cyclic schedule administered after surgical treatment of endometriomas were included, with pain and endometrioma recurrence rates as the primary outcomes. Three reviewers independently assessed methodology and extracted data from selected studies. The primary outcomes were considered pain recurrence (evaluated separately for dysmenorrhea, noncyclic chronic pelvic pain, and dyspareunia) and endometrioma recurrence evaluated at ultrasonography. Dichotomous outcomes from each study were expressed as risk ratio (RR) with a 95% confidence interval (CI). Three randomized clinical trials and 1 prospective controlled cohort study were included, for a total of 557 patients with endometriosis, 343 patients of whom had ovarian endometriomas completing the assigned treatment and follow-up. Lower recurrence rates for dysmenorrhea were obtained with a continuous schedule (RR, 0.24; 95% CI, 0.06-0.91; P = .04). Nonsignificant differences were present for chronic pelvic pain and dyspareunia. A continuous oral contraceptive schedule was associated with a nonsignificant reduction of cyst recurrence rates compared with a cyclic schedule (RR, 0.54; 95% CI, 0.28-1.05; P = .07). A continuous oral contraceptive regimen, as opposed to a cyclic regimen, may be suggested after surgery for endometriomas because of lower dysmenorrhea recurrence rates. Due to the small number and small sample sizes of the included studies, further randomized clinical trials are needed to confirm the findings of the present systematic review. Also, outcomes related to patient satisfaction and quality of life should be addressed.


Doughnut phenomenon in bone scintigraphy.
A doughnut phenomenon was observed in bone scintigraphy in three cases of metastatic thyroid carcinoma to the skull. Additional dynamic and static studies were performed to evaluate the relative roles of vascularity, vessel permeability and new bone formation in producing this phenomenon. This was accomplished through the use of 99mTc-diphosphonate, 99mTc-labeled red blood cells and 99mTc-pertechnetate. The important factor was found to be a new bone formation. Late uptake of diphosphonate was shown only in the periphery of the lesion, where formation of new bone takes place. This occurred even in the presence of increased vascularity and vessel permeability in the lesion as a whole.


Cancer cachexia--a clinical entity?
A number of metabolic abnormalities associated with cancer may interfere with normal starvation-adaption to alter the pattern of tissue loss in individuals with cancer. The cachexia thus produced is said to differ from simple under-nutrition and may therefore be detectable using simple clinical methods. Seven measurements commonly used for nutritional assessment were made before treatment in 72 patients with cancer and 89 patients who were undernourished but cancer-free. Body weight as a percentage of ideal weight, arm muscle circumference, triceps skinfold thickness, plasma protein, albumin and transferrin and the peripheral blood lymphocyte count did not differ significantly between the groups. If body composition in cancer cachexia differs from that found in comparable simple undernutrition then such variations are not detectable by a standard nutritional assessment.


[Stereotaxy and robotics for ablation - toy or tool?].
CLINICAL AND METHODOLOGICAL ISSUES: Ablative technologies allow local curative tumor treatment by thermal tissue damage. An important prerequisite is the coverage of all tumor cells. Tumor size is the most important limiting factor. The drawbacks of conventional computed tomography/ultrasound/magnetic resonance imaging (CT/US/MRI) guided radiofrequency ablation (RFA) are the absence of planning software, imprecise probe placement, imprecise control of probe placement and the ablation zone as well as the lack of reliability and reproducibility. Stereotactic and robot-assisted systems allow planning of multiple probe positions based on CT/MRI and positron emission tomography (PET) planning data. The probes can be precisely placed according to the coordinates of the image datasets. The 1 and 3 year survival rates after stereotactic RFA (SRFA) of cholangiocellular carcinoma were 91% and 70% respectively and the median overall survival was 60 months. After SRFA of 189 colorectal liver metastases in 63 patients there was no significant difference in local recurrence rates between tumors < 3 cm (17.7%), 3-5 cm (11.1%) and > 5 cm (17.4%). The median overall survival was 33.2 months and the 1, 3, and 5 year overall survival rates after SRFA in patients with resectable colorectal cancer were 92%, 66% and 48%, respectively. In our opinion the excellent and, to a large extent user-independent results justify the increased efforts in time and costs especially for the treatment of patients with large and irregular tumors. Stereotaxy and robotics are valuable tools for effective tumor ablation especially of large tumors and are likely to gain in importance in the next few years.


Primary fusion in worker's compensation intraarticular calcaneus fracture. Prospective study of 169 consecutive cases.
To study the results of reconstruction and primary fusion in worker's compensation intraarticular calcaneus fractures. We carried out a prospective study of 169 acute intraarticular calcaneus fractures treated by reconstruction and primary fusion with the minimally invasive Vira® system, in severe calcaneus fractures. The evaluation was performed by clinical, radiological and biomechanical analysis. AOFAS score averaged 77.26 points at the end of follow up. Forty-two cases (24.9%) obtained excellent results, 108 (63.9%) good, 12 (7.1%) mild and 7 (4.1%) poor. The improvement in Börder's angle after surgery was significant (p = 0.05) and this did not vary during the follow up. Subtalar arthrodesis was achieved in all cases and only three cases needed bone grafting. Five major post-surgical complications were observed, and one deep infection in a case of open Gustilo Grade III fracture. In the kinetic study, the support time of the operated foot was lower than that of the contralateral foot (p<0.21). The axial force of the heel contact and the single limb support of the operated foot reduced the toe-off axial forces. In the foot with arthrodesis the posterior forces increased (p <0.01). The pressures were lower in the region of the heel and the mid-foot and in the external part of the forefoot, and increased in the big toe. Calcaneal workplace injuries are challenging to treat. Primary subtalar fusion with a minimally invasive method allows rapid recovery for these patients with a satisfactory clinical, functional and radiological outcome.


[Helping the "hard-core" smokers].
Smoking cessation specialists are frequently confronted with smokers who have great difficulty in stopping smoking, and who are either motivated to stop or are forced to stop for health, economic or statutory reasons. These smokers are composed of a mixed population but they have in common a heavy dependence on tobacco and a significant level of cigarette consumption. They are exposed to serious morbidity induced by their uncontrollable smoking. Other factors unfavourable to the attempt to stop smoking are often present: anxiety-depressive disorders, socioeconomic difficulties or the use of psychoactive substances. They constitute a priority target for smoking cessation clinics, which must optimise and diversify proposals to improve their interventions. This review describes these highly dependent smokers unable to stop, and suggests medical treatments and therapeutic combinations to assist the practitioners trying to help the "hard-core" smokers.


Target cues are not required for the guidance of sympathetic preganglionic axons.
The patterns of preganglionic projections in the avian sympathetic system are highly specific. Preganglionic axons arising from the T1 spinal segment, for example, project predominantly in the rostral direction, whereas T4 preganglionic axons project predominantly in the caudal direction. Moreover, each ganglion receives projections from a distinct set of spinal cord segments. These patterns are established during embryonic life and are correct from the outset. In this study, I have examined the patterns of preganglionic projections in embryos deprived of their target ganglia through removal of the neural crest prior to its migration to form the sympathetic ganglia. My results indicate that preganglionic axons of target-deprived embryos still project in the proper directions and terminate in the presumptive target regions. Target cues are thus not required for the guidance of preganglionic axons.


Vaccination Against Human Papillomavirus.
Who should receive human papillomavirus (HPV) vaccination? Catch-up vaccination is now recommended for all persons through age 26 years. For persons aged 27 through 45 years, clinicians and patients should now jointly decide whether HPV vaccination is appropriate. Routine HPV vaccination at age 11 or 12 years (or as early as age 9 years) continues to be recommended.


The Clinical Genome and Ancestry Report: An interactive web application for prioritizing clinically implicated variants from genome sequencing data with ancestry composition.
Genome sequencing is positioned as a routine clinical work-up for diverse clinical conditions. A commonly used approach to highlight candidate variants with potential clinical implication is to search over locus- and gene-centric knowledge databases. Most web-based applications allow a federated query across diverse databases for a single variant; however, sifting through a large number of genomic variants with combination of filtering criteria is a substantial challenge. Here we describe the Clinical Genome and Ancestry Report (CGAR), an interactive web application developed to follow clinical interpretation workflows by organizing variants into seven categories: (1) reported disease-associated variants, (2) rare- and high-impact variants in putative disease-associated genes, (3) secondary findings that the American College of Medical Genetics and Genomics recommends reporting back to patients, (4) actionable pharmacogenomic variants, (5) focused reports for candidate genes, (6) de novo variant candidates for trio analysis, and (7) germline and somatic variants implicated in cancer risk, diagnosis, treatment and prognosis. For each variant, a comprehensive list of external links to variant-centric and phenotype databases are provided. Furthermore, genotype-derived ancestral composition is used to highlight allele frequencies from a matched population since some disease-associated variants show a wide variation between populations. CGAR is an open-source software and is available at https://tom.tch.harvard.edu/apps/cgar/.


Examining metastatic behavior within 3D bioprinted vasculature for the validation of a 3D computational flow model.
Understanding the dynamics of circulating tumor cell (CTC) behavior within the vasculature has remained an elusive goal in cancer biology. To elucidate the contribution of hydrodynamics in determining sites of CTC vascular colonization, the physical forces affecting these cells must be evaluated in a highly controlled manner. To this end, we have bioprinted endothelialized vascular beds and perfused these constructs with metastatic mammary gland cells under physiological flow rates. By pairing these in vitro devices with an advanced computational flow model, we found that the bioprinted analog was readily capable of evaluating the accuracy and integrated complexity of a computational flow model, while also highlighting the discrete contribution of hydrodynamics in vascular colonization. This intersection of these two technologies, bioprinting and computational simulation, is a key demonstration in the establishment of an experimentation pipeline for the understanding of complex biophysical events.


Two Unrelated Burkitt Lymphomas Seven Years Apart in a Patient With X-Linked Lymphoproliferative Disease Type 1 (XLP1).
We describe a rare case of a male child with X-linked lymphoproliferative disease type 1 (XLP1) who presented with Burkitt lymphoma (BL) when he was 6 years old, achieved a complete response to therapy, and developed a second BL after seven years. Diagnostic H&E stained slides and ancillary studies were reviewed for both lymphomas. B-cell clonality by PCR and SNP array studies were performed on both specimens. Both lymphomas were Epstein-Barr virus (EBV) negative. Flow cytometry showed λ light chain restriction in the initial BL and κ light chain restriction in the subsequent BL. B-cell clonality testing indicated that the two lymphomas are not clonally related. SNP array analysis of the second BL showed genomic changes that were not present in the first BL. These results confirm that these two tumors represent unrelated BLs. Pathologists and clinicians should be aware that second lymphomas in XLP1 patients may represent new neoplasms rather than late relapses.


Cyclodextrins improving the physicochemical and pharmacological properties of antidepressant drugs: a patent review.
Depression is a serious mood disorder and is one of the most common mental illnesses. Despite the availability of several classes of antidepressants, a substantial percentage of patients are unresponsive to these drugs, which have a slow onset of action in addition to producing undesirable side effects. Some scientific evidence suggests that cyclodextrins (CDs) can improve the physicochemical and pharmacological profile of antidepressant drugs (ADDs). The purpose of this paper is to disclose current data technology prospects involving antidepressant drugs and cyclodextrins. We conducted a patent review to evaluate the antidepressive activity of the compounds complexed in CDs, and we analyzed whether these complexes improved their physicochemical properties and pharmacological action. The present review used 8 specialized patent databases for patent research, using the term 'cyclodextrin' combined with 'antidepressive agents' and its related terms. We found 608 patents. In the end, considering the inclusion criteria, 27 patents reporting the benefits of complexation of ADDs with CDs were included. The use of CDs can be considered an important tool for the optimization of physicochemical and pharmacological properties of ADDs, such as stability, solubility and bioavailability.


SKIN-COUPLED PVDF MICROPHONES FOR NONINVASIVE VASCULAR BLOOD SOUND MONITORING.
Vascular access is the "Achilles Heel" of hemodialysis, as maintaining high flow characteristics (access patency) is critical to achieving efficient dialysis treatment. Thus, monitoring of vascular access is essential for maintaining long-term dialysis success. Blood sounds change in the presence of stenosis and can be analyzed digitally as phonoangiograms (PAGs) to determine changes in hemodynamic flow. We propose a multi-channel PAG recording sensor suitable for rapid, non-invasive vascular access monitoring. Here we present the initial design and characterization of sensors appropriate for recording PAGs from the skin surface. An optimized sensor size and backing material was selected to improve sensitivity and to provide a neutral frequency response. The sensor performance was finally compared with a conventional stethoscope on a controlled blood flow stenosis benchtop phantom.


Executive failure hypothesis explains the trait-level association between motivation and mind wandering.
Mind wandering (MW) is commonly observable in daily life. Early studies established an association between motivation and MW at the trait level using a questionnaire survey. Considering that the mechanism of state-level association between them is known, this study was conducted to replicate the trait-level association and determine its possible mechanisms. Two independent samples were analysed using several questionnaires, which included motivation and MW. General one- and multi-dimensional scales were administered for both variables. Besides the successful replication of the significant association between motivation and MW at the trait level, we found that people with low levels of executive function experience high rates of spontaneous MW. This finding indicates that the underlying mechanism of trait-level association is the executive failure hypothesis, which postulates that a failure of executive control during task-related objectives evokes MW. Further, the motivation-MW relationship exhibits a different psychological basis at the state and trait levels.


Osteopontin knockdown suppresses tumorigenicity of human metastatic breast carcinoma, MDA-MB-435.
Elevated expression of osteopontin (OPN), a secreted phosphoglycoprotein, is frequently associated with many transformed cell lines. Various studies suggest that OPN may contribute to tumor progression as well as metastasis in multiple tumor types. High levels of OPN have been reported in patients with metastatic cancers, including breast. We found that the expression of OPN corroborates with the aggressive phenotype of the breast cancer cells i.e. the expression of OPN is acquired as the breast cancer cells become more aggressive. To assess the role(s) of OPN in breast carcinoma, expression of endogenous OPN was knocked down in metastatic MDA-MB-435 human breast carcinoma cells using RNA interference. We targeted multiple regions of the OPN transcript for RNA interference, along with 'scrambled' and 'non-targeting siRNA pool' controls to distinguish between target-specific and potential off-target effects including interferon-response gene (PeIF2-alpha) induction. The OPN knockdown by shRNA suppressed tumor take in immunocompromised mice. The 'silenced' cells also showed significantly lower invasion and migration in modified Boyden chamber assays and reduced ability to grow in soft agar. Thus, in addition to the widely reported roles of OPN in late stages of tumor progression, these results provide functional evidence that OPN contributes to breast tumor growth as well.


Products of consultation-liaison psychiatry.
The products of consultation and liaison activities differ considerably and require distinct marketing strategies. Whereas consultation results directly in improved patient care, the products of psychiatric liaison typically consist of less easily measured changes in nonpsychiatric physicians' attitudes and skills. The difficulty of demonstrating the benefits of liaison psychiatry as presently conceived suggest that its future course lies in the active collaboration of liaison psychiatrists in specialized treatment programs.


The Hierarchy of Case Priority (HiCaP):- A proposed method for case prioritisation in digital forensic laboratories.
The need for digital forensic science (DFS) services has grown due to widespread and consistent engagement with technology by members of society. Whilst digital evidence often plays an important role in many inquiries, available investigative resources have failed to keep pace with such demand for them. As a result, the use case prioritisation models for backlog/workload management are of increasing importance to ensure the effective deployment of laboratory resources. This work focuses on the concept of case prioritisation in a digital forensic laboratory setting, following the submission of exhibits for examination, where this workflow is described. The challenges of case management and prioritisation in laboratories are discussed, with both 'case acceptance' and 'case prioritisation' procedures explained. Finally, the 'Hierarchy of Case Priority' (HiCaP) - a transparent, risk-based approach for the prioritisation of cases for examination, is proposed and described using examples.


Optimization of continuous TAG production by Nannochloropsis gaditana in solar-nitrogen-limited culture.
Nitrogen limitation and changing solar conditions are both known to affect triacylglycerol (TAG) production in microalgae. This study investigates the optimization of TAG production with a continuous nitrogen-limited culture of Nannochloropsis gaditana in simulated day-night cycles (DNc). The effect of DNc was first investigated in nitrogen-deprived condition (i.e., batch culture), emphasizing a significant change in mechanical resistance of the strain during the night. The concept of released TAG, which shows how much of the TAG produced is actually recovered in the downstream stages, that is, after cell disruption, was shown here of interest. For a maximum released TAG, the optimum harvesting time was suggested as being 4 h into the night period, which minimizes the losses due to a too great cell mechanical resistance. The protocol for continuous nitrogen-limited culture was then optimized, and a continuous nitrogen addition was compared to a pulsed-addition. For the latter, nitrogen was supplied in a single pulse at the beginning of the light periods, while the bulk medium was supplied separately at a slow but constant dilution rate of 0.005h-1$0.005\,{{\rm{h}}}^{-1}$ . The pulse dose was calculated after the study of nitrogen consumption and TAG production/consumption during the DNc. The estimated released TAG for the pulsed-addition of 1.4 × 10-3 kg/m2 d found significantly higher than the one achieved in batch culture (0.3 × 10-3 kg/m2 d) but lower than for continuous nitrogen addition which obtained the highest released TAG of 3×10-3kg∕m2 d$3\times 1{0}^{-3}\,\text{kg}\unicode{x02215}{{\rm{m}}}^{2}\unicode{x0200A}{\rm{d}}$ .


Chimeric Antigen Receptor T-Cell Therapy in Metastatic Castrate-Resistant Prostate Cancer.
Prostate cancer is the most commonly diagnosed solid-organ cancer amongst males worldwide. Metastatic castrate-resistant prostate cancer (mCRPC) is a rapidly fatal end-sequelae of prostate cancer. Therapeutic options for men with mCRPC are limited and are not curative in nature. The recent development of chimeric antigen receptor T-cell (CAR-T) therapy has revolutionised the treatment of treatment-resistant haematological malignancies, and several studies are underway investigating the utility of this technology in the treatment of solid tumours. In this review, we evaluate the current treatment options for men with mCRPC as well as the current landscape of preclinical and clinical trials of CAR-T cell therapy against prostate cancer. We also appraise the various prostate cancer-specific tumour-associated antigens that may be targeted by CAR-T cell technology. Finally, we examine the potential translational barriers of CAR-T cell therapy in solid tumours. Despite preclinical success, preliminary clinical trials in men with prostate cancer have had limited efficacy. Therefore, further clinically translatable preclinical models are required to enhance the understanding of the role of this investigational therapeutic in men with mCRPC. In the era of precision medicine, tailored immunotherapy administered to men in a tumour-agnostic approach provides hope to a group of men who otherwise have few treatment options available.


Isolation of temperature-sensitive pantothenate kinase mutants of Salmonella typhimurium and mapping of the coaA gene.
Temperature-sensitive pantothenate kinase mutants of Salmonella typhimurium LT2 were selected by using the excretion of pantothenate at the nonpermissive temperature as a screening method. Thermolability of the pathothenate kinase activity in extracts of the mutants was demonstrated. The mutations were mapped at min 89 of the Salmonella chromosome, near rpoB, by transduction. As pantothenate kinase catalyzes the first step in the biosynthesis of coenzyme A from pantothenate, the new genetic locus has been designated coaA.


Update: High altitude pulmonary edema.
Recent high altitude studies with pulmonary artery (PA) catheterization and broncho-alveolar lavage (BAL) in early high altitude pulmonary edema(HAPE) have increased our understanding of the pathogenetic sequence in HAPE. High preceding PA and pulmonary capillary pressures lead to a non-inflammatory leak of the alveolar-capillary barrier with egress of red cells, plasma proteins and fluid into the alveolar space. The mechanisms accounting for an increased capillary pressure remain speculative. The concept that hypoxic pulmonary vasoconstriction (HPV) is uneven so that regions with less vasoconstriction are over-perfused and become edematous remains compelling but unproved. Also uncertain is the role and extent of pulmonary venoconstriction. With disruption of the normal alveolar-capillary barrier, some individuals may later develop a secondary inflammatory reaction. A high incidence of preceding or concurrent respiratory infection in children with HAPE has been used to support a causative role of inflammation in HAPE. However, alternatively even mild HPV may simply lower the threshold at which inflammation-mediated increases in alveolar capillary permeability cause significant fluid flux into the lung. Other major questions to be addressed in future research are: 1.) What is the mechanism of exaggerated hypoxic pulmonary vasoconstriction? Is there a link to primary pulmonary hypertension? Several observations suggest that susceptibility to HAPE is associated with endothelial dysfunction in pulmonary vessels. This has not yet been studied adequately. 2.) What is the nature of the leak? Is there structural damage, i. e. stress failure, or does stretch cause opening of pores? 3.) What is the pathophysiologic significance of a decreased sodium and water clearance across alveolar epithelial cells in hypoxia? 4.) What is the role of exercise? Do HAPE-susceptible individuals develop pulmonary edema when exposed to hypoxia without exercise? Answers to these questions will increase our understanding of the pathophysiology of HAPE and also better focus research on the genetic basis of susceptibility to HAPE.


Metabolic risk factors in children with kidney stone disease.
The evaluation of metabolic risk factor in children with renal stone disease is the basis of medical treatment aimed at preventing recurrent stone events and the growth of preexisting calculi. In this retrospective study, we evaluated the metabolic risk factors and clinical and family histories of 90 children with kidney stone disease who had been referred to our institution and subjected to clinical tests using a standardized protocol. The mean age of our pediatric patients was 10.7 years, and the male:female ratio was 1.14:1.0. Biochemical abnormalities were found in 84.4% of all cases. A single urine metabolic risk factor was present in 52.2% (n = 47) of the patients, and multiple risk factors were present in the remaining 31.1% (n = 28). Idiopathic hypercalciuria (alone or in combination) and hypocitraturia (alone or in combination) were the most frequent risk factors identified in 40 and 37.8% of these patients, respectively. Renal colic or unspecified abdominal pain were the most frequent forms of presentation (76.9%), with 97.5% of stones located in the upper urinary tract. In most patients, stone disease was confirmed by renal ultrasonography (77%). A positive family history in first-degree and second-degree relatives was found in 46.2 and 32.5% of the cases, respectively. We conclude that specific urine metabolic risk factors are found in most children with kidney stones and that hypocitraturia is as frequent as hypercalciuria. Very often there is a positive family history of renal stone disease in first- and second-degree relatives.


Evolution of Risk Calculators and the Dawn of Artificial Intelligence in Predicting Patient Complications.
Risk calculators are an underused tool for surgeons and trainees when determining and communicating surgical risk. We summarize some of the more common risk calculators and discuss their evolution and limitations. We also describe artificial intelligence models, which have the potential to help clinicians better understand and use risk assessment.


Randomized controlled trial of cefazolin monotherapy versus cefazolin plus azithromycin single dose prophylaxis for cesarean deliveries: A developing country's perspective.
To compare the efficacy of pre-incision intravenous single doses of cefazolin versus cefazolin plus azithromycin as an antibiotic prophylaxis in cesarean delivery (CD). This was a single-center, double blind, randomized controlled trial conducted in the PGIMER, Chandigarh. 200 women undergoing elective/emergency cesarean section were randomized. Group A received single dose of cefazolin plus placebo while Group B received single dose of cefazolin plus azithromycin. Primary outcome evaluated was occurrence of surgical site infections (SSI); secondary outcomes included incidence of febrile morbidity, UTI, endometritis, neonatal outcome, total cost of antibiotics, and duration of hospital stay in both the study arms. Descriptive statistics and χ2 tests were used for analysis of the data. There was an overall significant reduction in the incidence of SSI (15% vs 3%; P = 0.03), endometritis (8% vs 2%; P = 0.048), and post-operative febrile morbidity (17% vs 3%; P = 0.001) with the addition of azithromycin to cefazolin. Duration of hospital stay was almost two days lesser for the cefazolin plus azithromycin group. Subgroup analysis of patients with SSI showed the age, duration of ruptured membranes, and type of anesthesia as important predictors of infection rate. Study observed statistically significant reduction in requirement of additional post operative antibiotics, phototherapy for neonates, hospital stay and cost of therapy in cefazolin plus azithromycin group (P < 0.05). Tertiary care hospitals in developing countries such as India can opt for the cefazolin plus azithromycin as antimicrobial prophylaxis during CD to maximize the efficacy as well as for decreasing the cost burden of postoperative infections.


Characterization of a Pt mirror to be used to deflect synchrotron radiation beam onto Langmuir monolayers.
A homemade mirror for X-rays has been built to prepare a diffraction beamline for liquid surface diffraction and scattering measurements. This simple approach is in operation at the XRD2 bending-magnet beamline at the Brazilian Synchrotron Light Laboratory.


Cimetidine protection against alpha-amanitin hepatotoxicity in mice: a potential model for the treatment of Amanita phalloides poisoning.
The ingestion of the mushroom Amanita phalloides is associated with hepatic necrosis appearing clinically two to three days after ingestion. The mechanism of this toxicity is unknown, and no reliable antidote is available. Because of the similarity of Amanita poisoning to other toxins affecting cytochrome P450, we investigated the use of cimetidine (as a P450 cytochrome inhibitor) as an antidote against a primary toxin of the mushroom alpha-amanitin. Mice injected with alpha-amanitin and given cimetidine either prophylactically or within six hours showed histologic protection from the hepatic damage seen in control mice. Control animals displayed significant mitochondrial changes when examined by electron-microscopy, while the mitochondria of the cimetidine-treated animals were preserved, suggesting a possible site for toxin action. Further trials will be necessary before treatment of human cases with cimetidine is indicated.


Dopaminergic responsivity in alcoholism: trait, state, or residual marker?
In order to classify neuroendocrine abnormalities in alcohol-dependent patients as trait, state, or residual markers, growth hormone (GH) secretion was assessed longitudinally. GH secretion, stimulated by the dopaminergic agonist apomorphine, was evaluated in 21 alcohol-dependent patients (16 men, five women) and 10 healthy comparison subjects (eight men, two women). The patients were tested during early withdrawal, after 8 days of abstinence, and after 3 months. Patients who relapsed within 3 months (N = 8) showed significantly less GH secretion in all neuroendocrine tests than did either the patients who abstained from ethanol consumption for 6 months (N = 13) or the healthy comparison subjects. The relapsers and abstainers did not differ significantly in any of their clinical or pathophysiological data, in the severity of their withdrawal symptoms, or in antecedent or concomitant illnesses associated with alcoholism. GH blunting appears to be a residual marker of clinical relevance in alcoholism.


Parathyroid hormone is associated with decreased fat mass in young healthy women.
To investigate the relationship of parathyroid hormone (PTH) with dietary calcium and changes in body composition. Cross-sectional and 1-year longitudinal trial. Normal-weight young women (age: 18-31), 155 subjects analyzed at baseline, and data for 41 subjects analyzed prospectively between baseline and 12 months. Levels of fasting serum calcium and PTH, intakes of calcium (3-day diet records), and total body weight and body composition (dual energy X-ray absorptiometry). Baseline dietary calcium, regardless of whether unadjusted or adjusted for energy intake, did not predict baseline levels of fasting serum PTH. Change in dietary calcium also did not predict change in serum PTH. However, log PTH was significantly correlated with body fat mass (R = 0.27), but not lean mass at baseline (n = 155), independent of serum calcium (corrected R = 0.25). Further, 12-month changes (n = 41) in log PTH positively predicted the 12-month change in body weight (R = 0.32) and body fat (R = 0.32), but not lean mass even when controlled for age or change in serum calcium. Fasting serum PTH was associated with increased fat mass, in both cross-sectional and prospective analysis. Thus, serum PTH may play a role in the regulation of body fat mass in young women.


The human immunodeficiency virus (HIV)-type 1 coreceptor CXCR-4 (fusin) is preferentially expressed on the more immature CD34+ hematopoietic stem cells.
In synergy with the CD4 antigen, the chemokine receptor CXCR-4 functions as a coreceptor for T-cell-tropic HIV-1 strains. Using two- and three-color immunofluorescence analysis, we examined the expression of CXCR-4 on CD34+ cells in 21 samples obtained from leukapheresis (LP) products of cancer patients who underwent G-CSF-supported cytotoxic chemotherapy. In addition, eight samples from bone marrow (BM) were obtained. CXCR-4 was expressed on the surface of CD34+ cells from samples of both hematopoietic sources. The mean proportion of CD34+/CXCR-4+ cells from LP products was 1.7-fold greater in comparison with those from bone marrow (65.9+/-4.1% vs. 37.5+/-8.6% [+/- SEM], p < 0.05). For an intraindividual comparison, LP products and bone marrow from six patients were obtained on the same day, confirming the significantly greater proportion of CD34+ cells coexpressing CXCR-4 cells in LP products. In order to examine whether the CXCR-4 expression was related to the stage of maturation and differentiation of CD34+ cells, six samples from LP products and four samples from bone marrow were assessed using three-color immunofluorescence analysis. We found that the subset of CD34+/CD38low and CD34+/HLA-DRlow cells representing a population of more immature progenitor cells were brightly positive for CXCR-4, while there was a decrease in the level of CXCR-4 expression in the population of CD34+/HLA-DRbright and CD34+/CD38bright cells. Based on the assessment of ten LP products, we found that the mean proportion of CD34+ cells coexpressing CD4 and CXCR-4 was 6.2+/-2.3% [+/- SEM], suggesting that a small population of CD34+ cells are, in principle, susceptible for an infection with T-cell-tropic HIV-1 strains. In conclusion, our data suggest that CXCR-4 is present on the surface of hematopoietic progenitor cells--particularly more primitive CD34+ cells. CXCR-4 could play a role in the homing of CD34+ cells to stromal elements of the bone marrow via its natural ligand stromal-derived factor-1 (SDF-1).


Effects of surface finish on indentation modulus and hardness of dental composite restoratives.
The depth-sensing micro-indentation testing was recently introduced for the characterization of dental composites. One of the critical issues raised was the possible influence of surface finish on material properties. The aim of this study was to investigate the effects of surface finish on the indentation modulus and micro-hardness of resin-based dental composite materials. The materials used included minifill (Z100, 3M ESPE), microfill (A110, 3M ESPE) and poly-acid modified (F2000, 3M ESPE) composites. The specimens were polished successively using SiC grinding papers of different grit size and diamond suspensions to achieve varying surface roughness. The arithmetic mean of the roughness (R(a)) was measured using profilometry. In the depth-sensing micro-indentation test, specimens (n=7) were indented to 10N with Vickers indenter and the load-displacement (P-h) data was obtained using a universal testing system. The indentation modulus (E(in)) and hardness (H) were then computed using the developed analytical solutions. Data was analyzed using ANOVA/post-hoc Scheffe's test at significance level 0.05. The polished specimens had surface roughness ranging from 0.02 to 0.81 microm. The roughness of F2000 was significantly higher than A110 and Z100. The E(in) and H for Z100 ranged from 14.02 to 14.83GPa and 1.18 to 1.27 GPa, respectively. E(in) for F2000 and A110 ranged from 12.25 to 13.82 GPa and 5.26 to 5.52 GPa and hardness ranged from 0.89 to 0.98 GPa and 0.52 to 0.55 GPa, respectively. The indentation modulus and hardness of dental composite restoratives were independent of the surface finish provided indenter penetration is sufficiently deep (h(max)/R(a)>30).


Resource partitioning in the rhizosphere by inoculated Bacillus spp. towards growth stimulation of wheat and suppression of wild oat (Avena fatua L.) weed.
Common wheat (Triticum aestivum L.) is one of the most important agricultural crop, which provides direct source of food for humans. Besides abiotic stresses, weeds pose a significant challenge to successful crop production. Avena fatua (wild oat) is one of the most common damaging grass weed, which causes 17-62% losses in yield of winter wheat. Excessive use of herbicides to control wild oat has resulted in serious environmental and human health hazards. Therefore, biological control of weeds is required to cope up with the increasing food demand and to attain self-sustainability. In this study, eighty eight rhizobacterial isolates were isolated from rhizosphere soil samples collected from Rewari and Hisar districts. After screening of the isolates, only thirty isolates showed in vitro antagonistic and herbicidal activities. The selected antagonistic isolates were further tested for production of IAA and ALA, and utilization of ACC. Bacterial isolates BWA18 and RWA52 produced 53.80 and 19.18 ug ml-1 IAA, respectively and high ALA production was shown by isolates HCA3 and RCA3. Five isolates i.e., BWA20, BWA23, BWA29, BWA38 and RCA3 showed significant ACC utilization. Inoculation of selected bacterial isolates BWA18, RWA69 and SYB101 showed significant increase in root dry weight (RDW) and shoot dry weight (SDW) of wheat plants under pot house conditions, and decreased RDW and SDW of A. fatua weed as compared to RDF-amended uninoculated soil at 25 DAS (days after sowing). Bacterial isolates RWA69 and SYB101 caused significant increase in RDW and SDW of wheat growth at 50 DAS, whereas their inoculation decreased RDW and SDW of A. fatua. Thus, seed bacterization with bacterial isolates RWA52, RWA69 and SYB101 caused significant increase in RDW and SDW of wheat, whereas their inoculation caused significant decrease in RDW and SDW of A. fatua. The best performing bacterial isolates RWA52 and RWA69 were identified as Bacillus siamensis and Bacillus endophyticus using 16S rRNA analysis. The promising rhizobacterial isolates could further be tested for the bioherbicidal activity and plant growth promotion effects under field conditions before their use as bioherbicides.


Analytical Approaches to Uncover Genetic Associations for Rare Outcomes: Lessons from West Nile Neuroinvasive Disease.
West Nile viral infection causes severe neuroinvasive disease in less than 1% of infected humans. There are no targeted therapeutics for this serious and potentially fatal disease, and to date no vaccine has been approved for humans. With climate change expected to result in rising incidence of West Nile and other related vector-borne viral infections, there is an increasing need to identify those at risk for serious disease and potential leads for therapeutic and vaccine development. Genetic variation, particularly in genes whose products are either directly or indirectly connected to immune response to infections, is a critical avenue of investigation to identify those at higher risk of clinically apparent West Nile infection. Given the small percent of infections that progress to severe disease and the relatively low numbers of reported infections, it is challenging to conduct well-powered studies to identify genetic factors associated with more severe outcomes. In this chapter, we outline several approaches with the objective to take full advantage of all available data in order to identify genetic factors which lead to increased risk of severe West Nile neuroinvasive disease. These methods are generalizable to other conditions with limited cohort size and rare outcomes.


Ultrastructural studies of developing pulmonary alveolar septal elastin.
Eighteen fetal lambs have been studied at six different gestational ages ranging from 90 days to term. At 90 days gestation, only the microfibrillar component of elastin is present in pulmonary alveolar septal elastic fibers. The amorphous component appears at 110 days and gradually increases in amount, relative to microfibrils. At term (150 days), the composition of the pulmonary alveolar septal elastin resembles that of the adult ewe.


Is autonomic support of arterial blood pressure related to habitual exercise status in healthy men?
We determined if the tonic autonomic nervous system (ANS) contribution to arterial blood pressure (BP) maintenance in humans is related to habitual endurance exercise status. Twenty-three healthy young (age 18-31 years) males, 11 endurance exercise-trained and 12 untrained, were studied. Maximal oxygen consumption was higher (P < 0.001) and resting heart rate and body fatness were lower (P < 0.05) in the exercise-trained men. Plasma noradrenaline concentrations and BP decreased from baseline levels in response to ganglionic blockade (intravenous trimethaphan) in both groups (all P < 0.001). The absolute (Delta mmHg: systolic = -35 +/- 2 vs. -32 +/- 4; diastolic = -13 +/- 2 vs. -10 +/- 2; mean = -21 +/- 2 vs. -17 +/- 3) and relative (Delta%: systolic = -35 +/- 2 vs. -31 +/- 3; diastolic = -26 +/- 3 vs. -20 +/- 3; mean = -31 +/- 2 vs. -26 +/- 3) decreases in BP were not significantly different between the endurance-trained and untrained men. There were no significant group differences in the heart rate, stroke volume, cardiac output or systemic vascular resistance (conductance) responses to trimethaphan. Systemic vascular alpha-adrenergic sensitivity (slope of the increase in mean BP with incremental phenylephrine infusion during ganglionic blockade) also did not differ in the two groups (endurance-trained: 3.2 +/- 0.5; untrained: 3.2 +/- 0.7 mmHg (ng phenylephrine)(-1) (ml plasma)(-1)). In the pooled sample, the decrease in mean BP during trimethaphan was related to baseline and changes in plasma noradrenaline concentrations (r = 0.58-0.65, P < 0.001) and alpha-adrenergic sensitivity (r = 0.49, P < 0.02). Our results suggest that the endurance exercise-trained state is not obviously associated with altered ANS support of BP in healthy young men. Basal sympathetic nervous system (SNS) activity and alpha-adrenergic vascular sensitivity are significant physiological correlates of ANS support of BP in this population.


Characteristics and outcomes of patients hospitalized with COVID-19 at a tertiary hospital in Nigeria.
Data regarding the features and outcomes of hospitalized COVID-19 patients in Africa are increasingly available. To describe socio-demographic, clinical and laboratory characteristics and outcomes of COVID-19 patients. A cross-sectional study of 86 adult patients hospitalized with COVID-19 between March and November 2020. Characteristics were described in survivors and non-survivors. Mean age was 60.9±16.1 years, 53(61.6%) were male. Co-morbidities were found in 77(89.5%) patients. On severity, 6(7%) were mild, 23(26.7%) moderate, 51(59.3%) severe and 6(7%) critical. Oxygen saturation and respiratory rate were 71±22% and 38±11/minute in non-survivors and 90±7% and 31±7/minute in survivors respectively (p<0.001, p<0.001)). Overall mortality was 47.7% with no death among patients with mild disease and deaths in all patients with critical disease. Duration of hospitalization was 2.0(1.0-4.5) days in those who died and 12(7.0-15.0) days in those who survived (p<0.001). Of the 42 patients that received dexamethasone, 11(26.2%) died, while 31(73.8%) survived (p=<0.001). Most of the patients had co-morbidities and there was high mortality in patients with severe and critical COVID-19. Mean oxygen saturation was low and respiratory rate high overall. Factors associated with mortality included: Significantly greater hypoxia and tachypnea, less dexamethasone use and shorter hospitalization.


The effectiveness of a multimedia intervention on parents' knowledge and use of vehicle safety systems for children.
Trauma due to road crashes is the leading cause of death and injury in Canadian children younger than 14 years, despite mandatory use of vehicle restraints since 1977. A multisite intervention study using a pretest-posttest design was conducted in four Ontario cities to test the effectiveness of an educational program on parents' knowledge of safety system use for children (0-12 years) 6 weeks following the educational intervention. The sample consisted of 418 families who reported on 732 children. Results indicated that knowledge increased significantly following the intervention. Use of professional sources of information was an important factor that is linked with increased parents' knowledge.


Pneumatic tactile stimulus delivery system for studying brain responses evoked by active finger touch with fMRI.
Primates use their hands to actively touch objects and collect information. To study tactile information processing, it is important for participants to experience tactile stimuli through active touch while monitoring brain activities. Here, we developed a pneumatic tactile stimulus delivery system (pTDS) that delivers various tactile stimuli on a programmed schedule and allows voluntary finger touches during MRI scanning. The pTDS uses a pneumatic actuator to move tactile stimuli and place them in a finger hole. A photosensor detects the time when an index finger touches a tactile stimulus, enabling the analysis of the touch-elicited brain responses. We examined brain responses while the participants actively touched braille objects presented by the pTDS. BOLD responses during tactile perception were significantly stronger in a finger touch area of the contralateral somatosensory cortex compared with that of visual perception. The pTDS enables MR studies of brain mechanisms for tactile processes through natural finger touch.


Plant-Derived Terpenoids: A Promising Tool in the Fight against Melanoma.
Melanoma is responsible for the highest number of skin cancer-caused deaths worldwide. Despite the numerous melanoma-treating options, the fight against it remains challenging, mainly due to its great heterogeneity and plasticity, as well as the high toxicity of standard drugs. Plant-derived terpenoids are a group of plant defense molecules that have been proven effective in killing many different types of cancer cells, both in in vitro experiments and in vivo models. In this review, we focus on recent results in the search for plant terpenoids with anti-melanoma activity. We also report on the synergistic action of combining terpenoids with other plant-derived substances, MAP kinase inhibitors, or radiation. Additionally, we present examples of terpenoid-loaded nanoparticle carriers as anti-melanoma agents that have increased permeation through the cancer tissue.


DeepNeuro: an open-source deep learning toolbox for neuroimaging.
Translating deep learning research from theory into clinical practice has unique challenges, specifically in the field of neuroimaging. In this paper, we present DeepNeuro, a Python-based deep learning framework that puts deep neural networks for neuroimaging into practical usage with a minimum of friction during implementation. We show how this framework can be used to design deep learning pipelines that can load and preprocess data, design and train various neural network architectures, and evaluate and visualize the results of trained networks on evaluation data. We present a way of reproducibly packaging data pre- and postprocessing functions common in the neuroimaging community, which facilitates consistent performance of networks across variable users, institutions, and scanners. We show how deep learning pipelines created with DeepNeuro can be concisely packaged into shareable Docker and Singularity containers with user-friendly command-line interfaces.


The interaction of cholinomimetics, peptides and compounds 48/80 on histamine secretion from the mast cell.
The mechanism of selective, non-immunological histamine release from mast cells, caused by various endogenous substances, is not clearly understood. Since in vivo experiments indicate that the local control of secretory cells is influenced by acetylcholine and peptides, we investigated whether the secretion of histamine is similarly regulated in the mast cell. Experiments were performed with peritoneal cavity cell suspensions (PCS) of the rat. The endogenous polypeptide substance P, compound 48/80 and a non-hydrolysable cholinomimetic agonist, carbachol, were used. The concentrations of the drugs were kept low to avoid non-specific histamine release caused by morphological damage of mast cells. It was found that: (1) carbachol (2 X 10(-5) M) did not release histamine from PCS, (2) substance P (6.5 X 10(-6) M) released histamine and this effect was increased by the addition of carbachol (2 X 10(-5) M); the effect of carbachol was inhibited by atropine , (3) carbachol (2 X 10(-5)M) did not increase histamine release caused by compound 48/80 (0.02 micrograms/ml). From these experiments it may be concluded that activation of peptidergic receptor(s) can cause histamine release from mast cells and that muscarinic agents may be involved in the regulation of the(se) receptor(s).


TNF-insulin crosstalk at the transcription factor GATA6 is revealed by a model that links signaling and transcriptomic data tensors.
Signal transduction networks coordinate transcriptional programs activated by diverse extracellular stimuli, such as growth factors and cytokines. Cells receive multiple stimuli simultaneously, and mapping how activation of the integrated signaling network affects gene expression is a challenge. We stimulated colon adenocarcinoma cells with various combinations of the cytokine tumor necrosis factor (TNF) and the growth factors insulin and epidermal growth factor (EGF) to investigate signal integration and transcriptional crosstalk. We quantitatively linked the proteomic and transcriptomic data sets by implementing a structured computational approach called tensor partial least squares regression. This statistical model accurately predicted transcriptional signatures from signaling arising from single and combined stimuli and also predicted time-dependent contributions of signaling events. Specifically, the model predicted that an early-phase, AKT-associated signal downstream of insulin repressed a set of transcripts induced by TNF. Through bioinformatics and cell-based experiments, we identified the AKT-repressed signal as glycogen synthase kinase 3 (GSK3)-catalyzed phosphorylation of Ser(37) on the long form of the transcription factor GATA6. Phosphorylation of GATA6 on Ser(37) promoted its degradation, thereby preventing GATA6 from repressing transcripts that are induced by TNF and attenuated by insulin. Our analysis showed that predictive tensor modeling of proteomic and transcriptomic data sets can uncover pathway crosstalk that produces specific patterns of gene expression in cells receiving multiple stimuli.


Identification of nucleotide substitution in gene encoding [LeuA3]insulin in third Japanese family.
We previously described a new case of abnormal insulinemia in Japan. In one allele, nucleotide-sequence analysis revealed a substitution in the codon for the third position of insulin A chain (GTG----TTG), causing [LeuA3]insulin. This point mutation is the same as that found in insulin Wakayama. In this family, the mutant insulin allele cosegregated with an 850-base pair PvuII allele of the hypervariable region 5'-flanking the insulin gene.


The ceramide activated protein phosphatase Sit4 impairs sphingolipid dynamics, mitochondrial function and lifespan in a yeast model of Niemann-Pick type C1.
The Niemann-Pick type C is a rare neurodegenerative disease that results from loss-of-function point mutations in NPC1 or NPC2, which affect the homeostasis of sphingolipids and sterols in human cells. We have previously shown that yeast lacking Ncr1, the orthologue of human NPC1 protein, display a premature ageing phenotype and higher sensitivity to oxidative stress associated with mitochondrial dysfunctions and accumulation of long chain bases. In this study, a lipidomic analysis revealed specific changes in the levels of ceramide species in ncr1Δ cells, including decreases in dihydroceramides and increases in phytoceramides. Moreover, the activation of Sit4, a ceramide-activated protein phosphatase, increased in ncr1Δ cells. Deletion of SIT4 or CDC55, its regulatory subunit, increased the chronological lifespan and hydrogen peroxide resistance of ncr1Δ cells and suppressed its mitochondrial defects. Notably, Sch9 and Pkh1-mediated phosphorylation of Sch9 decreased significantly in ncr1Δsit4Δ cells. These results suggest that phytoceramide accumulation and Sit4-dependent signaling mediate the mitochondrial dysfunction and shortened lifespan in the yeast model of Niemann-Pick type C1, in part through modulation of the Pkh1-Sch9 pathway.


Quantitative estimation of salicylic acid and its metabolites by thin-layer densitometry.
A rapid thin-layer densitometric method for the quantitative determination of salicylic acid and its metabolites in urine or plasma is described. The method is specific and very sensitive. Nanogram quantities of salicylic acid and its metabolites, both free and conjugated, may be estimated. Known metabolites, as well as the newly described gentisuric acid, were estimated quantitatively in urine from a patient treated with aspirin.


A simple procedure for immunocytochemical- and silver-staining of endocrine cells in the same section.
A new simple staining and restaining procedure for peptide hormone/biogenic amine containing endocrine cells is described. With this, immunocytochemistry and restaining with a silver technique can be applied to the same histological section for the identification and characterization of endocrine cells.


High seroprevalence of HCV in the Abruzzo Region, Italy: results on a large sample from opt-out pre-surgical screening.
Available estimates of the prevalence of chronic HCV infection in Italy are quite conflicting, varying from 1.5 to 22.5%, with an apparent north to south gradient. As Direct Acting Antivirals are expensive, both National and local governmental Agencies are in urgent need of detailed and reliable estimates of HCV patients to be treated, nationwide and in each district. We investigated the prevalence of anti-HCV antibodies in a large unselected sample of surgical patients providing consent to in-hospital opt-out pre-surgical HCV screening, at two hospitals from the Abruzzo Region, Italy. Data were retrieved for 55,533 screened patients (4.1% of the total population in the Abruzzo Region), admitted in the Orthopedic and General Surgery wards of Pescara and Teramo Hospitals from 1999 to 2014. The prevalence of anti-HCV antibodies was 4.4% in the total sample. HCV-positive patients had a mean age of 63.8 ± 19.9 years; 49.2% were males. From 1999 to 2014, the prevalence of HCV antibodies decreased from 5.4% to 4.1%; at both sites, however, two age-related-peaks were evidenced, the first among patients aged 30-49 years, the second among those older than 70 years. Statistical analyses confirmed a significant trend to decrease over time and a higher prevalence in Pescara and among males (all p < 0.01). Data retrieved from opt-out pre-surgical screening programs may allow inexpensive and easy-to-perform estimates of HCV seroprevalence from large samples of unselected patients with a well-defined provenience, which may turn useful for future treatment resource allocation.


Protective effect of vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes.
Organophosphate (OP) pesticides such as dimethoate and malathion intoxication has been shown to produce oxidative stress due to the generation of free radicals and alter the antioxidant defense system in erythrocytes. It is possible that vitamin E being present at the cell membrane site may prevent OP-induced oxidative damage. In the present study, rats were pretreated orally with vitamin E (250 mg/kg body wt, twice a week for 6 weeks) prior to oral administration of a single low dose of dimethoate and/or malathion (0.01% LD(50)). The result showed that treatment with OP increased lipid peroxidation (LPO) in erythrocytes, however, vitamin E pretreated rats administered OP's showed decreased LPO in erythrocytes. The increase in the activities of superoxide dismutase (SOD) and catalase (CAT) and total-SH content in erythrocytes from dimethoate and/or malathion treated rats as compared to control appears to be a response towards increased oxidative stress. Vitamin E pretreated animals administered OP's showed a lowering in these parameters as compared to OP treated rats which indicates that vitamin E provide protection against OP-induced oxidative stress. The glutathione-S-transferase (GST) activity in erythrocytes was inhibited in OP intoxicated rats which partially recovered in vitamin E pretreated animals administered OP's. Inhibition in erythrocyte and serum acetylcholinesterase (AChE) activity was not relieved in vitamin E pretreated rats administered OP's probably due to the competitive nature of enzyme inhibition by OP's. The results show that vitamin E may amelierate OP-induced oxidative stress by decreasing LPO and altering antioxidant defense system in erthrocytes.