Management of Connective Tissue Disease-Associated Interstitial Lung Disease.
The presence of interstitial lung disease (ILD) negatively affects prognosis among patients with an underlying connective tissue disease (CTD). The initial approach to care should determine whether the CTD-ILD needs pharmacologic treatment or not. There is little direct evidence to guide who and how to treat. At present, any severe, active, and/or progressive ILD should be pharmacologically treated. Immunosuppressants and/or corticosteroids are the mainstay of pharmacologic therapy for all CTD-ILDs, whereas antifibrotics may be beneficial in some scenarios. A comprehensive and multidisciplinary approach to management is also an important aspect of patient care.


Fine-Tuning Mitochondrial Dysfunction and Reductive Carboxylation.
Metabolic processes within cells are dynamically interconnected. If mitochondria become defective, cells must rewire their metabolism to survive. Here we highlight recent work by Gaude et al. that used a tunable model of mitochondrial dysfunction combined with metabolic tracing and in silico analyses to define these compensatory pathways.


Privacy-preserving storage of sequenced genomic data.
The current and future applications of genomic data may raise ethical and privacy concerns. Processing and storing of this data introduce a risk of abuse by potential offenders since the human genome contains sensitive personal information. For this reason, we have developed a privacy-preserving method, named Varlock providing secure storage of sequenced genomic data. We used a public set of population allele frequencies to mask the personal alleles detected in genomic reads. Each personal allele described by the public set is masked by a randomly selected population allele with respect to its frequency. Masked alleles are preserved in an encrypted confidential file that can be shared in whole or in part using public-key cryptography. Our method masked the personal variants and introduced new variants detected in a personal masked genome. Alternative alleles with lower population frequency were masked and introduced more often. We performed a joint PCA analysis of personal and masked VCFs, showing that the VCFs between the two groups cannot be trivially mapped. Moreover, the method is reversible and personal alleles in specific genomic regions can be unmasked on demand. Our method masks personal alleles within genomic reads while preserving valuable non-sensitive properties of sequenced DNA fragments for further research. Personal alleles in the desired genomic regions may be restored and shared with patients, clinics, and researchers. We suggest that the method can provide an additional security layer for storing and sharing of the raw aligned reads.


Biosynthesis of ferritin in rat hepatoma cells and rat livers. I. Synthesis and assembly of protein subunits of ferritin.
Cell fractions were prepared from ACI rat livers and from rat hepatoma cell clone M-5123-C1. Radioimmunoassays of ferritin and of its protein subunits in various cell fractions after biosynthetic labeling with [14C]leucine were done by means of ferritin-specific and subunit-specific rabbit antibody. In both ACI rat livers and M-5123-C1 hepatoma cells free polyribosomes synthesized approximately 81% of the protein subunits of ferritin, and membrane-bound polyribosomes synthesized the rest. In both polyribosomal fractions, [14C]leucine-labeled subunits were detected earlier than [14C]leucine-labeled ferritin and apoferritin (5 min as against 30 min after initiation of a pulse). Time sequence studies of the shifts of biosynthetically labeled subunits and ferritin through different cell compartments provided evidence for vectorial transport of subunits and of ferritin, the direction of transport being from the two polyribosomal systems to the smooth membrane compartment and to the cytosol.


Closed medial total subtalar joint dislocation without ankle fracture: a case report.
Total subtalar dislocation without fracture of the ankle is a rare clinical entity; it is usually due to a traumatic high-energy mechanism. Standard treatment is successful closed reduction under general anesthesia followed by non-weight bearing and ankle immobilization with a below-knee cast for 6 weeks. We present the case of a 30-year-old Moroccan woman who was involved in a road traffic accident. She subsequently received a radiological assessment that objectified a total subtalar dislocation without fracture of her ankle. She was immediately admitted to the operating theater where an immediate reduction was performed under sedation, and immobilization in a plaster boot was adopted for 8 weeks. The management of this traumatic lesion is discussed in the light of the literature. Medial subtalar dislocation is a rare dislocation and is not commonly seen as a sports injury because it requires transfer of a large amount of kinetic energy. The weaker talocalcaneal and talonavicular ligaments often bear the brunt of the energy and are more commonly disrupted, compared to the relatively stronger calcaneonavicular ligament. Urgent reduction is important, and closed reduction under general anesthesia is usually successful, often facilitated by keeping the knee in flexion to relax the gastrocnemius muscle. Long-term sequelae include talar avascular necrosis and osteochondral fracture, as well as chronic instability and pain.


Fetal manifestations and poor outcomes of congenital cytomegalovirus infections: possible candidates for intrauterine antiviral treatments.
This retrospective study was performed to reveal the natural history of cytomegalovirus (CMV) infected fetuses during the perinatal period and to find prenatal findings associated with poor outcomes. 33 neonates with CMV infection, born after 30 weeks of gestation, were registered from a total of 12 414 infants between 1995 and 2003. Maternal and neonatal medical records were reviewed regarding fetal growth; abdominal signs including ascites and hepatosplenomegaly; cerebral signs including ventriculomegaly, microcephaly, and calcification; and fetal heart rate monitoring, for signs which may have been detected by the standard obstetric ultrasonography. Univariate and multivariate analyses were performed to test for any associations between these manifestations and poor outcomes such as death and neurological damages. Among the 33 infants, 6 died, 10 developed neurological damage including cerebral palsy (n = 8), epilepsy (n = 5), and hearing difficulties (n = 5), and the remaining 17 were normal. After adjusting for ganciclovir treatment and gender, death was 40-fold more likely associated with infants having abdominal signs (OR 40, 95%CI 4.6-930) than those without abdominal signs. Similarly, poor outcomes (death or neurological damage) were more likely associated with infants having either abdominal or cerebral signs (OR 39, 95%CI 3.8-1323). Fetal growth restriction and non-reassuring fetal heart rate patterns were not significantly associated with poor outcomes. The absence of abdominal signs guarantees the infant's survival. The presence of abdominal or cerebral signs is associated with poor outcomes, suggesting that these fetuses are possible candidates to receive in-utero therapy of congenital CMV infection.


Fever and intracranial pressures.
The effect of fever upon intracranial pressures was determined in the rabbit and cat. In the unanesthetized rabbit and cat, cerebrospinal fluid (CSF) pressure was measured via direct cannulation of the lateral cerebral ventricle. Intracranial pressure (ICP) was measured in the rabbit by a subarachnoid screw technique. In all cases, intravenous administration of bacterial pyrogen extracted from Salmonella abortus equi resulted in significant differences from controls in physiological variables measured during the initial "chill" phase of the fever. There was an increase in body temperature, a fall in CSF or ICP pulse rate, an increase in pulse pressure amplitude, and a small increase in mean CSF or ICP. In addition, venous and arterial blood pressures increased significantly and, consistent with heat conservation, there was a fall in respiratory rate as well as cutaneous vasoconstriction in the ears. The arterial carbon dioxide tension was unchanged during the prodrome but fell significantly during the chill and flush phases and rose again during defervescence. The results suggest that in these animals there is a slight increase in pressures within the cranium during the "chill" phase of a pyrogen induced fever, resulting from changes occurring in many body systems during this phase of the fever.


Magnetic nanoparticle incorporated oleogel as iontophoretic drug delivery system.
In this article, we validated the use of electric current as an external stimulus to induce an enhancement of drug release from magnetic nanoparticle (MNP) incorporated organogels (magnetogel) under iontophoretic conditions. For this purpose, we adopted a simple, two-step synthesis route to incorporate magnetic (Fe3O4) nanoparticles (MNP) and ciprofloxacin hydrochloride within the network of a soybean oil-based oleogel using stearic acid as gelator. We fabricated a series of MNP incorporated oleogels by varying the wt% of MNPs while keeping a constant weight ratio of soybean oil:stearic acid. The microstructures of the magnetogels were analyzed in MNP concentration-dependent manner by optical microscopy, powder X-ray diffraction, FTIR, mechanical, and electrical studies. Detailed analysis of the electrical properties revealed that the gel sample with a maximum proportion of MNP (S4) allowed the maximum passage of current through it among all the compositions. Under the iontophoretic environment of the active condition, we observed nearly 2.5 fold increase in cumulative drug release in case of sample S4 compared to the corresponding passive condition. These observations suggested that in future, our magnetogel formulation can be further developed as AC field induced 'remote controlled' agent for therapeutic application.


Guided Web-Based Cognitive Behavior Therapy for Perfectionism: Results From Two Different Randomized Controlled Trials.
Perfectionism can become a debilitating condition that may negatively affect functioning in multiple areas, including mental health. Prior research has indicated that internet-based cognitive behavioral therapy can be beneficial, but few studies have included follow-up data. The objective of this study was to explore the outcomes at follow-up of internet-based cognitive behavioral therapy with guided self-help, delivered as 2 separate randomized controlled trials conducted in Sweden and the United Kingdom. In total, 120 participants randomly assigned to internet-based cognitive behavioral therapy were included in both intention-to-treat and completer analyses: 78 in the Swedish trial and 62 in the UK trial. The primary outcome measure was the Frost Multidimensional Perfectionism Scale, Concern over Mistakes subscale (FMPS CM). Secondary outcome measures varied between the trials and consisted of the Clinical Perfectionism Questionnaire (CPQ; both trials), the 9-item Patient Health Questionnaire (PHQ-9; Swedish trial), the 7-item Generalized Anxiety Disorder scale (GAD-7; Swedish trial), and the 21-item Depression Anxiety Stress Scale (DASS-21; UK trial). Follow-up occurred after 6 months for the UK trial and after 12 months for the Swedish trial. Analysis of covariance revealed a significant difference between pretreatment and follow-up in both studies. Intention-to-treat within-group Cohen <italic>d</italic> effect sizes were 1.21 (Swedish trial; 95% CI 0.86-1.54) and 1.24 (UK trial; 95% CI 0.85-1.62) for the FMPS CM. Furthermore, 29 (59%; Swedish trial) and 15 (43%; UK trial) of the participants met the criteria for recovery on the FMPS CM. Improvements were also significant for the CPQ, with effect sizes of 1.32 (Swedish trial; 95% CI 0.97-1.66) and 1.49 (UK trial; 95% CI 1.09-1.88); the PHQ-9, effect size 0.60 (95% CI 0.28-0.92); the GAD-7, effect size 0.67 (95% CI 0.34-0.99); and the DASS-21, effect size 0.50 (95% CI 0.13-0.85). The results are promising for the use of internet-based cognitive behavioral therapy as a way of targeting perfectionism, but the findings need to be replicated and include a comparison condition.


An unusual cause of intractable secondary dysmenorrhea.
Severe dysmenorrhea following sterilization may be due to the presence of a noncommunicating horn of a bicornuate uterus. Ultrasound examination should be made before a functional basis is considered for such pain.


The finite element method as a research and teaching tool in the analysis of local skin flaps.
The finite element analysis (FEA) is a recently introduced method in biomechanics that permits modeling of complex structures considering them as an aggregate of small elements. Skin flaps are highly suggested to be amenable to the continuum mechanic laws that underly the development of FEA. A combination of "large deformation analysis," based on FEA with the criteria for skin flap selection, was attempted. Serial defects were experimentally created on piglet skin stripes, which were consequently covered through designing appropriate flaps. Skin samples were modeled after the development of a computer FEA program and they were scanned by incorporating their photographs. On the graphic interfaces the flap movement, the closure of the defect, and the whole deformation were found to match with the skin stripe postincisional alterations. This work permits the prediction and offers planning guides for different skin reconstructions.


Optimization of replacing pork back fat with grape seed oil and rice bran fiber for reduced-fat meat emulsion systems.
The effects of reducing pork fat levels from 30% to 20% and partially substituting the pork fat with a mix of grape seed oil (0%, 5%, 10% and 15%) and 2% rice bran fiber were investigated based on chemical composition, cooking characteristics, physicochemical and textural properties, and viscosity of reduced-fat meat batters. For reduced-fat meat batters containing grape seed oil and rice bran fiber the moisture and ash contents, uncooked and cooked pH values, yellowness, cohesiveness, gumminess, chewiness, and sarcoplasmic protein solubility were higher than in the control samples. The reduced-fat samples with increasing grape seed oil concentrations had lower cooking loss, emulsion stability, and apparent viscosity. The incorporation of grape seed oil and rice bran fiber successfully reduced the animal fat content in the final products while improving other characteristics.


High-throughput sensor microtiter plate of new terbium complexes for the determination of anthracene in seawater.
A new fluorescent sensing microtiter plate (MTP) was developed for high sensitivity monitoring of anthracene in seawater samples. For this purpose, two ternary complexes of Tb(III) ions with dibenzoylmethane and neocuproine [Tb(DBM)2(MePhen)] or with dibenzoylmethane and bathocuproine [Tb(DBM)2(PhMePhen)] were synthesized. Elemental analysis, energy dispersive X-ray analysis, X-ray diffraction, infrared and ultraviolet-visible emission, and thermal analysis were conducted on the Tb(III) complexes. The limits of detection (DL) were 0.14 and 1.05 μmol L-1 for [Tb(DBM)2(MePhen)] and [Tb(DBM)2(PhMePhen)], respectively. [Tb(DBM)2(PhMePhen)] MTP is embedded in a membrane made of cellulose acetate. The first high-throughput anthracene sensor MTP, based on [Tb(DBM)2(PhMePhen)] sensor showed a linear range, from 0.2 to 20 μmol L-1. [Tb(DBM)2(PhMePhen)] MTP was validated for accurate and precise monitoring of anthracene using gas chromatography. The selectivity of the [Tb(DBM)2(PhMePhen)] MTP toward anthracene was examined. The data indicated that [Tb(DBM)2(PhMePhen)] MTP is suitable for rapid and direct detection of anthracene.


Selection Maintains Protein Interactome Resilience in the Long-Term Evolution Experiment with Escherichia coli.
Most cellular functions are carried out by a dynamic network of interacting proteins. An open question is whether the network properties of protein interactomes represent phenotypes under natural selection. One proposal is that protein interactomes have evolved to be resilient, such that they tend to maintain connectivity when proteins are removed from the network. This hypothesis predicts that interactome resilience should be maintained by natural selection during long-term experimental evolution. I tested this prediction by modeling the evolution of protein-protein interaction (PPI) networks in Lenski's long-term evolution experiment with Escherichia coli (LTEE). In this test, I removed proteins affected by nonsense, insertion, deletion, and transposon mutations in evolved LTEE strains, and measured the resilience of the resulting networks. I compared the rate of change of network resilience in each LTEE population to the rate of change of network resilience for corresponding randomized networks. The evolved PPI networks are significantly more resilient than networks in which random proteins have been deleted. Moreover, the evolved networks are generally more resilient than networks in which the random deletion of proteins was restricted to those disrupted in LTEE. These results suggest that evolution in the LTEE has favored PPI networks that are, on average, more resilient than expected from the genetic variation across the evolved strains. My findings therefore support the hypothesis that selection maintains protein interactome resilience over evolutionary time.


Differential effects of proinflammatory cytokines on cell death and ER stress in insulin-secreting INS1E cells and the involvement of nitric oxide.
Proinflammatory cytokines produced by immune cells destroy pancreatic beta cells in type 1 diabetes. The aim of this study was to investigate the cytokine network and its effects in insulin-secreting cells. INS1E cells were exposed to different combinations of proinflammatory cytokines. Cytokine toxicity was estimated by MTT assay and caspase activation measurements. The NFκB-iNOS pathway was analyzed by a SEAP reporter gene assay, Western-blotting and nitrite measurements. Gene expression analyses of ER stress markers, Chop and Bip, were performed by real-time RT-PCR. Cytokines tested in this study, namely IL-1β, TNFα and IFNγ, had deleterious effects on beta cell viability. The most potent toxicity exhibited IL-1β and its combinations with other cytokines. The toxic effects of IL-1β towards cell viability, caspase activation and iNOS activity were dependent on nitric oxide and abolished by an iNOS blocker. IL-1β was the strongest inducer of the NFκB activation. An iNOS blocker inhibited IL-1β-mediated NFκB activation in the first, initial phase of cytokine action, but did not affect significantly NFκB activation after prolonged incubation. Interestingly iNOS protein expression was induced predominantly by IL-1β and decreased in the presence of an iNOS blocker in the case of a short time exposure. The changes in the expression of ER stress markers were also almost exclusively dependent on the IL-1β presence and counteracted by iNOS blockade. Thus cytokine-induced beta cell death is primarily IL-1β mediated with a NO-independent enhancement by TNFα and IFNγ. The deleterious effects on cell viability and function are crucially dependent on IL-1β-induced nitric oxide formation.


Following Trypanosoma cruzi RPA-DNA Interaction Using Fluorescent In Situ Hybridization Coupled with Immunofluorescence (FISH/IF).
Fluorescent in situ hybridization coupled with immunofluorescence (FISH/IF) is an assay that has been widely used to study DNA-protein interactions. The technique is based on the use of a fluorescent nucleic acid probe and fluorescent antibodies to reveal the localization of a DNA sequence and a specific protein in the cell. The interaction can be inferred by the quantification of the co-localization between the protein and the DNA. Here, we describe a detailed FISH/IF methodology that our group used to study RPA-telomere interaction in the pathogenic protozoa parasite Trypanosoma cruzi.


Effect of thoracotomy upon the carotid sinus control of regional hemodynamics in the dog.
The effects of bilateral thoracotomy upon the operating characteristics of the carotid sinus reflex were examined in 11 vagotomized dogs. Pressure-flow relationships in the aorta, celiac, mesenteric, renal, and femoral arteries were studied as a function of pressure in the isolated carotid sinuses. These relationships were analyzed to give the carotid sinus reflex operating point values and sensitivities of pressure, flow, impedance, and power for each regional vascular bed. Following thoracotomy there was a significant fall in the closed-loop operating point pressure (CLOP) ( p less than 0.005). The operating point values and sensitivities for each regional flow and resistance together with the corresponding mean and oscillatory kinetic and pressure power were not significantly altered by thoracotomy. Renal characteristic impedance at the operating point significantly increased following thoracotomy (p less than 0.05), as did the aortic characteristic impedance for carotid sinus pressures (CSP) above the closed-loop operating point. The operating sensitivity of renal characteristic impedance was decreased (p less than 0.05). Possible mechanisms underlying these changes are discussed.


Biological properties and molecular characterization of beet chlorosis virus (BChV).
Two distinct viruses belonging to the Polerovirus genus, in the family Luteoviridae, have been described as being able to induce mild yellowing on sugar beet: Beet mild yellowing virus (BMYV) and more recently, beet chlorosis virus (BChV). We have analysed biological properties and molecular organisation of two strains of BChV, one collected in England and the second from California. The biological data suggested that BChV displayed a narrower host range compared to BMYV and Beet western yellows virus lettuce isolate (BWYV). The complete genomic RNA sequence of the American isolate BChV-California and the European isolate BChV-2a showed a genetic organisation and expression typical of other Polerovirus members including 6 open reading frames (ORFs). Interspecific and intraspecific phylogenetic studies suggested that BChV arose by recombination events between a Polerovirus-like ancestor donating P0 and the replicase complex and either a BMYV or a BWYV progenitor providing the 3' ORFs [3, 4 and 5]. The 5'- and 3'-parts of the BChV genome have evolved differently in the two continents, possibly due to different selection pressures to allow adaptation to the various environments, hosts and vectors. BChV is a distinct species of the Polerovirus genus.


Microencapsulation of bioactive nanoparticles.
Supported or modified enzymes in the form of mobile nanoparticles are designed for enhanced activities and stabilities; however, their practical operations are dwarfed due to their tiny size which always makes recycling an arduous task and a potential risk to the environment. To overcome such drawbacks, this chapter describes a method for the preparation of a new form of microcapsules, possessing single-cavity compartments and nano-pores in the shell, to encage nanoparticle-based biocatalysts and form cell-like microreactors (CLMRs). The encaged nanoscale catalysts are maintained their high activities as in a bulk-phase solution, while they could be handled as materials of sizes hundreds-fold larger.


Mutagens and carcinogens in size-classified air particulates of a northern Italian town.
This research was designed to examine the presence of mutagenic/carcinogenic compounds in urban airborne particulate matter in relation to particles aerodynamic size. Inhalable (< 10 microns) airborne particulate (PM-10) was collected at a low traffic site in an industrialized Northern Italian town, using a high volume sampler equipped with a cascade impactor for particles fractionation. The organic extracts of different fractions were examined for mutagenicity in Salmonella typhimurium strains TA98 and TA98/1,8-DNP6 using the microsuspension procedure, and for polycyclic aromatic hydrocarbons (PAHs) content by gas chromatography. Size fractionated particles were also analysed for heavy metals (Fe, Mn, Zn, Pb, Cu, Cd, Cr, Ni, V) using plasma spectrophotometry. The results of mutagenicity and chemical analyses indicate that, at the site investigated, inhalable particulate was largely made of fine (< 0.5 micron) particles, which accounted for most of PAHs and mutagenicity. A similar pattern of distribution was found for heavy metals, which were relatively more abundant in small (< 1.5 microns) particles compared to coarser ones.


Assessment of alkaline stabilization processes in industrial waste streams using a model-based approach.
Chemical conditioning prior to disposal is a common practice in biotech companies to stabilize the biological waste generated during production. Nevertheless, the state of the art models used to analyze management strategies in water treatment systems (WTS) do not include the effect of high alkaline conditions during bio-solids processing. In this paper, the prediction capabilities of a novel model-based approach describing the effect of quicklime addition (CaO) on the waste streams of an industrial WTS is assessed. Two measuring campaigns were carried out taking samples of TSS, VSS and total/soluble COD, N, P, S and multiple metals before and after chemical stabilization, and dewatering under and overflow. Mass balances were set up and Sankey diagrams were generated to represent the occurrence, transformation and fate of the major compounds within the studied facility. A simulation model was used to predict plant at different locations. Next, a scenario analysis was carried out in order to assess potential alternatives to the current operational practice. The resulting mass balances show a mismatch between the system's input and output up to 17%. It was also possible to identify different types of compound-behavior depending on the effect that high pH induced on the soluble and particulate fractions: hydrolysis, precipitation and unaltered. Model predictions and measurements differed 9.6% (steady state) and 12.4% (dynamic state) respectively. Finally, in the scenario analysis, the model suggested that the change from quicklime to sodium hydroxide (NaOH) would increase the quantity of organics in the dewatered cake (+23%), but with a considerable increase in chemical consumption (+50%). The selective stabilization of the incoming streams has the lowest use of chemicals (-30%) and reduces the load of CODsol (-13%) and TNsol (-14%) recirculated to the water line of the WWTP.


Structural determinants of glutathione transferases with azathioprine activity identified by DNA shuffling of alpha class members.
A library of alpha class glutathione transferases (GSTs), composed of chimeric enzymes derived from human (A1-1, A2-2 and A3-3), bovine (A1-1) and rat (A2-2 and A3-3) cDNA sequences was constructed by the method of DNA shuffling. The GST variants were screened in bacterial lysates for activity with the immunosuppressive agent azathioprine, a prodrug that is transformed into its active form, 6-mercaptopurine, by reaction with the tripeptide glutathione catalyzed by GSTs. Important structural determinants for activity with azathioprine were recognized by means of primary structure analysis and activities of purified enzymes chosen from the screening. The amino acid sequences could be divided into 23 exchangeable segments on the basis of the primary structures of 45 chosen clones. Segments 2, 20, 21, and 22 were identified as primary determinants of the azathioprine activity representing two of the regions forming the substrate-binding H-site. Segments 21 and 22 are situated in the C-terminal helix characterizing alpha class GSTs, which is instrumental in their catalytic function. The study demonstrates the power of DNA shuffling in identifying segments of primary structure that are important for catalytic activity with a targeted substrate. GSTs in combination with azathioprine have potential as selectable markers for use in gene therapy. Knowledge of activity-determining segments in the structure is valuable in the protein engineering of glutathione transferase for enhanced or suppressed activity.


Objective assessment of resistance of female urethra to stress. A scale to establish degree of urethral incompetence.
During urethral pressure profile (UPP) studies under gradually increasing stress, the maximum urethral closure pressure decreases. This observation led to a simple urodynamic test which allows the estimation of the maximum stress (intensity of cough) tolerated by the urethra before leakage begins. The determination of this critical pressure forms the basis of the urethral incompetence scale. The urethra of a female patient can be characterized by the amplitude of this maximum stress (in cm H2O) the urethra can handle and still remain continent. The cough intensity scale has been divided into 5 segments, defining 5 degrees of urethral incompetence. The critical pressure, which determines the degree of urethral incompetence is a constant value for a given patient, provided the bladder volume is kept constant and the position of the patient during the examination is unchanged. This method establishes the functional status of the female urethra as far as its sphincteric capabilities are concerned under different stress conditions. It represents the degree of pressure transmission from the abdominal cavity to the proximal urethra during stress. It evaluates objectively the functional results of different surgical or medical therapeutic modalities to cure genuine stress incontinence. It allows a better selection of patients with unstable bladder and genuine stress incontinence who will more likely benefit from a retropubic cystourethropexy.


Risk of pelvic fractures in older women following pelvic irradiation.
Pelvic fractures, including hip fractures, are a major source of morbidity and mortality in older women. Although therapeutic pelvic irradiation could increase the risk of such fractures, this effect has not been studied. To determine if women who undergo pelvic irradiation for pelvic malignancies (anal, cervical, or rectal cancers) have a higher rate of pelvic fracture than women with pelvic malignancies who do not undergo irradiation. We conducted a retrospective cohort study using Surveillance, Epidemiology, and End Results (SEER) cancer registry data linked to Medicare claims data. A total of 6428 women aged 65 years and older diagnosed with pelvic malignancies from 1986 through 1999 were included. We compared results for women who did (n = 2855) vs did not (n = 3573) undergo radiation therapy. To assess the influence of selection bias, we also evaluated the effect of irradiation on osteoporotic fractures in nonirradiated sites (arm and spine). We evaluated the effect of irradiation on the incidence of pelvic fractures over time, and adjusted for potential confounders using a proportional hazards model. Women who underwent radiation therapy were more likely to have a pelvic fracture than women who did not undergo radiation therapy (cumulative 5-year fracture rate, 14.0% vs 7.5% in women with anal cancer, 8.2% vs 5.9% in women with cervical cancer, and 11.2% vs 8.7% in women with rectal cancer); the difference was statistically significant and most fractures (90%) were hip fractures. We controlled for potential confounders including age, race, cancer stage, and geographic location. The impact of irradiation varied by cancer site: treatment for anal cancer was associated with a higher risk of pelvic fractures (hazard ratio, 3.16; 95% confidence interval, 1.48-6.73); than for cervical cancer (hazard ratio, 1.66; 95% confidence interval, 1.06-2.59); or rectal cancer (hazard ratio, 1.65; 95% confidence interval, 1.33-2.05). No statistically significant difference was found in the rate of arm or spine fractures between the irradiated and nonirradiated groups (hazard ratio, 1.15; 95% confidence interval, 0.89-1.48). Pelvic irradiation substantially increases the risk of pelvic fractures in older women. Given the high baseline risk of pelvic fracture, this finding is of particular concern.


Oral and gastrointestinal sensing of dietary fat and appetite regulation in humans: modification by diet and obesity.
Dietary fat interacts with receptors in both the oral cavity and the gastrointestinal (GI) tract to regulate fat and energy intake. This review discusses recent developments in our understanding of the mechanisms underlying the effects of fat, through its digestive products, fatty acids (FAs), on GI function and energy intake, the role of oral and intestinal FA receptors, and the implications that changes in oral and small intestinal sensitivity in response to ingested fat may have for the development of obesity.


Cross-reactivity of hypervariable region 1 chimera of hepatitis C virus.
To analyze the amino acid sequences of hypervariable region 1 (HVR1) of HCV isolates in China and to construct a combinatorial chimeric HVR1 protein having a very broad high cross-reactivity. All of the published HVR1 sequences from China were collected and processed with a computer program. Several representative HVR1's sequences were formulated based on a consensus profile and homology within certain subdivision. A few reported HVR1 mimotope sequences were also included for a broader representation. All of them were cloned and expressed in E.coli. The cross-reactivity of the purified recombinant HVR1 antigens was tested by ELISA with a panel of sera from HCV infected patients in China. Some of them were further ligated together to form a combinatorial HVR1 chimera. Altogether 12 HVR1(s) were selected and expressed in E.coli and purified to homogeneity. All of these purified antigens showed some cross-reactivity with sera in a 27 HCV positive panel. Recombinant HVR1s of No. 1, 2, 4, and 8# showing broad cross-reactivities and complementarity with each other, were selected for the ligation elements. The chimera containing these 4 HVR1s was highly expressed in E.coli. The purified chimeric antigen could react not only with all the HCV antibody positive sera in the panel but also with 90/91 sera of HCV -infected patients. The chimeric antigen was shown to have a broad cross-reactivity. It may be helpful for solving the problem caused by high variability of HCV, and in the efforts for a novel vaccine against the virus.


A manual carotid compression technique to overcome difficult filter protection device retrieval during carotid artery stenting.
We investigated the incidence of embolic protection device retrieval difficulties at carotid artery stenting (CAS) with a closed-cell stent and demonstrated the usefulness of a manual carotid compression assist technique. Between July 2010 and October 2013, we performed 156 CAS procedures using self-expandable closed-cell stents. All procedures were performed with the aid of a filter design embolic protection device. We used FilterWire EZ in 118 procedures and SpiderFX in 38 procedures. The embolic protection device was usually retrieved by the accessory retrieval sheath after CAS. We applied a manual carotid compression technique when it was difficult to navigate the retrieval sheath through the deployed stent. We compared clinical outcomes in patients where simple retrieval was possible with patients where the manual carotid compression assisted technique was used for retrieval. Among the 156 CAS procedures, we encountered 12 (7.7%) where embolic protection device retrieval was hampered at the proximal stent terminus. Our manual carotid compression technique overcame this difficulty without eliciting neurologic events, artery dissection, or stent deformity. In patients undergoing closed-cell stent placement, embolic protection device retrieval difficulties may be encountered at the proximal stent terminus. Manual carotid compression assisted retrieval is an easy, readily available solution to overcome these difficulties.


Fatal Post COVID mRNA-Vaccine Associated Cerebral Ischemia.
Venous thromboses have been linked to several COVID-19 vaccines, but there is limited information on the Moderna vaccine's effect on the risk of arterial thrombosis. Here we describe a case of post-Moderna COVID-19 vaccination arterial infarct with vaccine-associated diffuse cortical edema that was complicated by refractory intracranial hypertension. 24 hrs after receiving her first dose of the Moderna COVID-19 vaccine, a 30-year-old female developed severe headache. Three weeks later she was admitted with subacute headache and confusion. Imaging initially showed scattered cortical thrombosis with an elevated opening pressure on lumbar puncture. An external ventricular drain was placed, but she continued to have elevated intracranial pressure. Ultimately, she required a hemicraniectomy, but intractable cerebral edema resulted in her death. Pathology was consistent with thrombosis and associated inflammatory response. Though correlational, her medical team surmised that the mRNA vaccine may have contributed to this presentation. The side effects of COVID-19 infection and vaccination are still incompletely understood. Though complications are rare, clinicians should be aware of presentations like this one.


Protective effect of a novel thromboxane antagonist, BAY-U3405, on canine myocardial damage after coronary artery occlusion and reperfusion.
The effects of a novel thromboxane antagonist, (3R)-3-(4-fluorophenylsulfonmido)-1,2,3,4-tetrahydro-9-carbazol epropanoic acid (BAY-U3405), on myocardial damage due to ischemia and reperfusion (added to accelerate the initiated injury) were studied in anesthetized dogs. The left anterior descending (LAD) coronary artery was occluded for 6 hours and reperfused for 30 minutes. BAY-U3405, 1 mg/kg, was injected intravenously 15 minutes after LAD occlusion, followed by continuous infusion of 10 mg/kg/hour starting at 30 minutes after occlusion. The drug had no hemodynamic effects. During the experiments 6 of 14 animals died of ventricular fibrillation (VF) in the placebo-vehicle controls (3 during occlusion and 3 during reperfusion); in the drug-treated group 5 of 13 dogs died of VF (all during occlusion none during reperfusion). This difference in total mortality and cause was not statistically significant. Reperfusion arrhythmias were largely suppressed by BAY-U3405: 201 +/- 43 versus 689 +/- 98 irregular beats during 30 minutes (p less than 0.001) in the experimental and control groups, respectively. Coronary collateral flow, obtained from a load-line analysis by measurement of retrograde coronary flow, and collateral index were similar in both groups. Therefore, BAY-U3405 did not alter collateral blood supply to the ischemic myocardium. Infarct size, determined with tetrazolium staining, was reduced by 65% (p less than 0.01) after its administration. These results suggest that thromboxane antagonism by BAY-U3405 may delay infarct expansion and reduce the frequency of ventricular arrhythmias during reperfusion of previously ischemic myocardium.


Balance of Gata3 and Ramp2 in hepatocytes regulates hepatic vascular reconstitution in postoperative liver regeneration.
Post-hepatectomy liver failure (PHLF) leads to poor prognosis in patients undergoing hepatectomy, with hepatic vascular reconstitution playing a critical role. However, the regulators of hepatic vascular reconstitution remain unclear. This study aimed to investigate the regulatory mechanisms of hepatic vascular reconstitution and identify biomarkers predicting PHLF in patients undergoing hepatectomy. Candidate genes that were associated with hepatic vascular reconstitution were screened using adeno-associated virus vectors in Alb-Cre-CRISPR/Cas9 mice subjected to partial hepatectomy. The biological activities of candidate genes were estimated using endothelial precursor transfusion and an associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) models. The level of candidates was detected in biopsies from patients undergoing ALPPS. Risk factors for PHLF were also screened using retrospective data. Downregulation of Gata3 and upregulation of Ramp2 in hepatocytes promoted the proliferation of liver sinusoidal endothelial cells (LSECs) and hepatic revascularization. Pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor A (VEGFA) played opposite roles in regulating the migration of endothelial precursors from bone marrow and the formation of new sinusoids after hepatectomy. Gata3 restricted endothelial cell function in patient-derived hepatic organoids, which was abrogated by a Gata3 inhibitor. Moreover, overexpression of Gata3 led to higher mortality in ALPPS mice, which was improved by a PEDF-neutralizing antibody. The expression of Gata3/RAMP and PEDF/VEGFA tended to have a negative correlation in patients undergoing ALPPS. A nomogram incorporating multiple factors, such as serum PEDF/VEGF index, was constructed and could efficiently predict the risk of PHLF. The balance of Gata3 and Ramp2 in hepatocytes regulates the proliferation of LSECs and hepatic revascularization via changes in the expression of PEDF and VEGFA, revealing potential targets for the prevention and treatment of PHLF. In this study, we show that the balance of Gata3 and Ramp2 in hepatocytes regulates hepatic vascular reconstitution by promoting a shift from pigment epithelium-derived factor (PEDF) to vascular endothelial growth factor A (VEGFA) expression during hepatectomy- or ALLPS (associating liver partition and portal vein ligation for staged hepatectomy)-induced liver regeneration. We also identified serum PEDF/VEGFA index as a potential predictor of post-hepatectomy liver failure in patients who underwent hepatectomy. This study improves our understanding of how hepatocytes contribute to liver regeneration and provides new targets for the prevention and treatment of post-hepatectomy liver failure.


High-performance liquid chromatography of coenzyme Q-related compounds and its application to biological materials.
A convenient and precise method for the separation and determination of coenzyme Q (CoQ)-related compounds (CoQ homologues, plastoquinone-9, ubichromenol-9, etc.) was developed using high-performance liquid chromatography (HPLC). All compounds tested were separated using a reverse-phase column with a suitable mobile phase and detected at a wavelength of 275 nm. CoQ extracts in plasma and erythrocytes were purified by thin-layer chromatography prior to HPLC analysis, but such purification was not necessary when determining CoQ in urine and tissues. Hydroquinone forms of CoQ existing in animal tissues were oxidized to the corresponding quinone forms with potassium hexacyanoferrate(III). This HPLC method was applied satisfactorily to the determination of the contents of CoQ homologues in human and animal samples. CoQ10 was the only homologue detected in human samples, and CoQ8, CoQ9 and CoQ10 were native homologues of CoQ in rat tissues. Ubichromenol-9 and plastoquinone-9 were not detected in these samples.


Microcanonical analysis of the kinetic method. the meaning of the "apparent entropy".
A rigorous analysis of the kinetic method is carried out using Rice-Ramsperger-Kassel-Marcus (RRKM) theory of microcanonical statistical unimolecular dissociation rates. The model employs a kinetics treatment appropriate for metastable ion dissociation. Proton-bound alkoxide dimer anions are used as model systems, with realistic vibrational and rotational parameters calculated by ab initio methods for the cluster ion and transition states leading to the competitive dissociation channels. The numerical simulations show that the kinetic method plots of ln(I2/I1) versus AAH are nearly linear but can exhibit significant curvature. The apparent entropy obtained in the extended kinetic method is not approximately equal to the thermodynamic entropy difference for dissociation, AAS(T), or for activation, deltadeltaS++(T), either at the effective temperature or at any fixed equilibrium temperature. Instead, the apparent entropy term can be related to the ratio of the microcanonical sum of states of the dissociation transition states for the kinetically selected internal energy of the dissociating ions.


Distal protection improved reperfusion and reduced left ventricular dysfunction in patients with acute myocardial infarction who had angioscopically defined ruptured plaque.
Distal protection, in the Saphenous Vein Graft Angioplasty Free of Emboli (SAFER) trial, is demonstrated to prevent distal embolism in the percutaneous coronary intervention of saphenous vein graft. However, in the Enhanced Myocardial Efficacy and Recovery by Aspiration of Liberated Debris (EMERALD) trial, it was not effective in the percutaneous coronary intervention of native coronary arteries in patients with acute myocardial infarction (AMI). We hypothesized that its effectiveness would be determined by lesion characteristics. Therefore, we classified the type of culprit lesion by angioscopy and examined its influence on the effectiveness of distal protection, comparing patients with AMI treated with and without distal protection. Consecutive patients with AMI treated without distal protection (n=110) from July 2000 to July 2002 and those treated with distal protection (n=81) from July 2002 to July 2004 were included. Patients in each group were subdivided according to whether or not they had angioscopically defined ruptured plaque at culprit lesion. Among those groups, incidence of no-reflow phenomenon, ST-segment resolution, myocardial blush grade, and left ventricular ejection fraction at 6 months were compared. Aspirated samples by distal protection were semiquantitatively and histologically analyzed and compared between patients with and without ruptured plaque. No-reflow phenomenon was most frequently (P<0.05) observed in patients with ruptured plaque treated without distal protection. ST-segment resolution (68+/-15% versus 40+/-21%, P<0.001), myocardial blush grade (2.6+/-0.5 versus 1.8+/-0.3, P<0.001), and left ventricular ejection fraction (47.2+/-6.7% versus 41.0+/-9.7%, P<0.01) were improved by distal protection among patients with ruptured plaque but not among patients without ruptured plaque. Aspirated samples >1 mm were detected more frequently (97.3% versus 78.5%, P<0.05) in patients with ruptured plaque than those without ruptured plaque. Histologically, aspirated samples contained plaque debris (95.3% versus 31.1%, P<0.05) more frequently in patients with ruptured plaque than in those without ruptured plaque. Distal protection reduced microcirculation damage and left ventricular dysfunction in patients with AMI who had angioscopically defined ruptured plaque. Distal embolization of plaque debris was detected more frequently in patients with ruptured plaque. These results suggest that microcirculation damage and left ventricular dysfunction are increased mainly by distal embolization of plaque debris rather than of thrombus.


Eye injuries in northern Sweden.
An analysis of 927 eye injuries treated at the University Hospital of Umeå, Northern Sweden, during a one-year period has been made. The estimated incidence was 8.1 per 1000 population. 86% of the injured were men. An overrepresentation of severe injuries were found in children and young adults, aged 10-19 years. 94.5% of the injuries were minor. The most common diagnosis was foreign body adherent to cornea or conjunctiva (40%). All chemical injuries were superficial. One-third of the sports injuries were caused by hockey-bockey and indoor bandy. Most of these were major injuries. The total cost of the injuries was assessed at SEK 1300,000. More information to the public concerning use of safety glasses and potential risks of eye trauma could reduce eye injury incidence.


RAPID3 (Routine Assessment of Patient Index Data 3) severity categories and response criteria: Similar results to DAS28 (Disease Activity Score) and CDAI (Clinical Disease Activity Index) in the RAPID 1 (Rheumatoid Arthritis Prevention of Structural Damage) clinical trial of certolizumab pegol.
To compare categories for activity/severity according to the Disease Activity Score 28-joint count (DAS28), the Clinical Disease Activity Index (CDAI), and the Routine Assessment of Patient Index Data 3 (RAPID3), an index without formal joint counts calculated in 5 versus >100 seconds, as well as the European League Against Rheumatism (EULAR)- DAS28 and the RAPID3 response criteria, in the Rheumatoid Arthritis Prevention of Structural Damage (RAPID 1) clinical trial of certolizumab pegol (CZP). Post hoc analyses were performed using correlations, cross-tabulations, and kappa statistics. Patients (treated with CZP plus methotrexate [MTX] or placebo plus MTX) were classified at baseline and at 52 weeks as high, moderate, low activity/severity or remission, according to the DAS28 (>5.1, >3.2 to ≤5.1, 2.6 to ≤3.2, <2.6 [total range 0-10]), the CDAI (>22, >10 to ≤22, >2.8 to ≤10, ≤2.8 [total range 0-76]), and RAPID3 (>12, >6 to ≤12, >3 to ≤6, ≤3 [total range 0-30]), as well as for good, moderate, and poor EULAR-DAS28 and proposed RAPID3 response criteria. All measures were correlated significantly: RAPID3 with DAS28 and CDAI (rho > 0.7), higher than erythrocyte sedimentation rate with C-reactive protein level (rho = 0.47). At 52 weeks, DAS28, CDAI, and RAPID3 low activity/remission was seen in 30%, 44%, and 42% of CZP-treated patients versus 3%, 7%, and 10% of control patients. Good, moderate, and poor EULAR-DAS28 responses were seen in 30%, 51%, and 19% of CZP-treated patients versus 3%, 28%, and 70% of control patients, and for RAPID3 in 39%, 30%, and 32% of CZP-treated patients versus 8%, 16%, and 76% of control patients. Kappa and weighted kappa values ranged from 0.36-0.53, indicating fair to moderate agreement. RAPID3, DAS28, and CDAI give similar results to distinguish CZP patients from controls in the RAPID 1 clinical trial. DAS28 is specific for clinical trials; RAPID3 appears pragmatically useful for usual care.


Antidepressant effects of sertraline associated with volume increases in dorsolateral prefrontal cortex.
Structural brain imaging has revealed differences in gray matter volume between depressed individuals and control subjects in key structures related to emotion. The aim of the present study was to assess within subjects whether regional changes in gray matter volume were observed over time in depressed patients treated with sertraline. Thirteen depressed patients were assigned to selective serotonin reuptake inhibitor (SSRI) treatment with sertraline. Structural magnetic resonance imaging (MRI) was performed at four time points over 12 weeks of treatment. Depressed individuals were compared to a control group of 10 subjects scanned at the same time points using voxel based morphometry and a statistical growth modeling technique. SSRI-treated patients showed a region of significantly increasing gray matter volume over time within the left dorsolateral prefrontal cortex (DLPFC) that specifically correlated with decreases in self-reported depression levels. No statistically significant changes were observed related to time in controls. This study included a small sample size and the method of analysis was capable of detecting only linear changes in volume. Effective antidepressant treatment with sertraline is associated with left DLPFC volume increases. These volume increases may reflect cortical architectural changes associated with top-down neuronal modulation of emotion.


Treatment processes for microplastics and nanoplastics in waters: State-of-the-art review.
In this work, established treatment processes for microplastics (MPs) and nanoplastics (NPs) in water as well as developed analytical techniques for evaluation of the operation of these processes were reviewed. In this regard, the strengths and limitations of different qualitative and quantitative techniques for the analysis of MPs and NPs in water treatment processes were first discussed. Afterward, the MPs and NPs treatment processes were categorized into the separation and degradation processes and the challenges and opportunities in their performance were analyzed. The evaluation of these processes revealed that the MPs or NPs removal efficiency of the separation and degradation processes could reach up to 99% and 90%, respectively. It can be concluded from this work that the combination of separation and degradation processes could be a promising approach to mineralize MPs and NPs in water with high efficiency.


Evidence for species differences in 'peripheral' benzodiazepine receptors: an autoradiographic study.
The presence, density and distribution of 'peripheral' benzodiazepine (BZ)-binding sites was investigated by autoradiography in the brains of rats, mice, guinea pigs and cats and in some areas of the dog and monkey brains, using [3H]Ro 5-4864 as a ligand. Marked interspecies differences were found in the distribution and densities of these sites. Rats and mice presented a low density of binding uniformly distributed throughout the brain with high densities concentrated in the ependyma, choroid plexus and olfactory nerve layer of the olfactory bulb. In contrast, guinea pig and cat brains presented relatively high concentrations of peripheral BZ binding sites throughout the grey matter ependyma and choroid plexus, but low in the olfactory bulb. Monkey and dog brains presented low densities of peripheral BZ binding sites, including the choroid plexus.


Nutmeg Essential Oil, Red Clover, and Liquorice Extracts Microencapsulation Method Selection for the Release of Active Compounds from Gel Tablets of Different Bases.
The current study presents the most suitable method for encapsulating nutmeg essential oil with liquorice and red clover. Two widely used methods, spray-drying and freeze-drying, were employed to find the most suitable for essential oil volatile compounds' protection. Results showed that freeze-dried capsules (LM) had a higher yield (85.34%) compared to the exact formulation of spray-dried microcapsules (SDM)-45.12%. All the antioxidant and total phenolic compounds' results obtained with the LM sample were significantly higher compared with SDM. LM microcapsules were incorporated in two different bases with no additional sugar (gelatin and pectin) for targeted release. Pectin tablets had firmer and harder texture properties, while gelatin tablets had a more elastic texture. There was a significant impact on texture changes caused by microcapsules. Microencapsulated essential oil with extracts can be used alone or in a gel base (pectin or gelatin, depending on user preferences). It could be an effective product to protect the active volatile compounds and regulate the release of active compounds and give a pleasant taste.


Melatonin protects the heart and pancreas by improving glucose homeostasis, oxidative stress, inflammation and apoptosis in T2DM-induced rats.
Cardiomyopathy and pancreatic injury are health issues associated with type 2 diabetes mellitus (T2DM) and are characterized by elevated oxidative stress, inflammation and apoptosis. Melatonin (MLT) is a hormone with multifunctional antioxidant activity. The protective effects of MLT on the heart and pancreas during the early development of diabetic cardiomyopathy and pancreatic injury were investigated in male Wistar rats with T2DM. MLT (10 mg/kg) was administered daily by gavage for 15 days after diabetic induction. Treatment of diabetic rats with MLT significantly normalized the levels of serum glucose, HbA1-c, and the lipid profile and improved the insulin levels and insulin resistance compared with diabetic rats, affirming its antidiabetic effect. MLT significantly prevented the development of oxidative stress and sustained the levels of glutathione and glutathione peroxidase activity in the heart and pancreas of diabetic animals, indicating its antioxidant capacity. Additionally, MLT prevented the increase in proinflammatory cytokines and expression of Bax, caspase-3 and P53. Furthermore, MLT enhanced the anti-inflammatory cytokine IL-10 and antiapoptotic protein Bcl-2. MLT controlled the levels of troponin T and creatine kinase-MB and lactate dehydrogenase activity, indicating its anti-inflammatory and antiapoptotic effects. Histological examinations confirmed the protective effects of MLT on T2DM-induced injury in the myocardium, pancreas and islets of Langerhans. In conclusion, the protective effects of melatonin on the heart and pancreas during the early development of T2DM are attributed to its antihyperglycemic, antilipidemic and antioxidant influences as well as its remarkable anti-inflammatory and antiapoptotic properties.


Neurodevelopment of the incentive network facilitates motivated behaviour from adolescence to adulthood.
The ability to enhance motivated performance through incentives is crucial to guide and ultimately optimise the outcome of goal-directed behaviour. It remains largely unclear how motivated behaviour and performance develops particularly across adolescence. Here, we used computational fMRI to assess how response speed and its underlying neural circuitry are modulated by reward and loss in a monetary incentive delay paradigm. We demonstrate that maturational fine-tuning of functional coupling within the cortico-striatal incentive circuitry from adolescence to adulthood facilitates the ability to enhance performance selectively for higher subjective values. Additionally, during feedback, we found developmental sex differences of striatal representations of reward prediction errors in an exploratory analysis. Our findings suggest that a reduced capacity to utilise subjective value for motivated behaviour in adolescence is rooted in immature information processing in the incentive system. This indicates that the neurocircuitry for coordination of incentivised, motivated cognitive control acts as a bottleneck for behavioural adjustments in adolescence.


High-resolution mapping reveals topologically distinct cellular pools of phosphatidylserine.
Phosphatidylserine (PS) plays a central role in cell signaling and in the biosynthesis of other lipids. To date, however, the subcellular distribution and transmembrane topology of this crucial phospholipid remain ill-defined. We transfected cells with a GFP-tagged C2 domain of lactadherin to detect by light and electron microscopy PS exposed on the cytosolic leaflet of the plasmalemma and organellar membranes. Cytoplasmically exposed PS was found to be clustered on the plasma membrane, and to be associated with caveolae, the trans-Golgi network, and endocytic organelles including intraluminal vesicles of multivesicular endosomes. This labeling pattern was compared with the total cellular distribution of PS as visualized using a novel on-section technique. These complementary methods revealed PS in the interior of the ER, Golgi complex, and mitochondria. These results indicate that PS in the lumenal monolayer of the ER and Golgi complex becomes exposed cytosolically at the trans-Golgi network. Transmembrane flipping of PS may contribute to the exit of cargo from the Golgi complex.


Involvement of ischemia-driven 5-lipoxygenase-resolvin-E1-chemokine like receptor-1 axis in the resolution of post-coronary artery bypass graft inflammation in coronary arteries.
Expression status of pro-resolvin lipid mediators (PLM) and receptors in the post-Coronary artery bypass grafting (CABG) coronary arteries are largely unknown. Here, we aim to investigate the expression of the enzymes involved in PLM synthesis and their receptors in the atherosclerotic post-CABG swine (AS) left anterior descending (LAD) compared to without CABG (LAD-AS), and in isolated coronary artery smooth muscle cells (CASMCs) cultured under ischemia. The arteries of interest were harvested from post-CABG atherosclerotic swine and the histomorphology and the expression status of key PLM mediators were quantified using immunostaining. Smooth muscle cells (SMCs) were cultured under ischemia and confirmed the expression on PLM mediators at transcript and protein level. The histomorphometric analysis revealed considerable alterations in the tissue architecture in LAD-CABG and LAD-AS arteries compared to control. PLM synthetic enzyme 5-lipoxygenases (5LO) was significantly upregulated in LAD-CABG and LAD-AS whereas the other enzymes including 12LO, 15LO, and cyclooxygenase-2, and the receptors including Chemokine like receptor 1 (ChemR23), 7-transmembrane G-protein coupled receptor-18 (GPCR18), GPCR120 were decreased in LAD-CABG than control. LO enzymes and PLM receptors were upregulated in ischemic CASMCs with respect to control. Western blot showed the upregulation of 5LO, and ChemR23. Additionally, higher level of resolvin-E1 (RvE1) was observed in ischemic control CASMCs which was decreased following reperfusion. These findings suggest that the CASMCs withstand the ischemia-triggered proinflammatory episodes by increasing the secretion of RvE1 mediated through 5LO and ChemR23 signaling.


Quantitation of free cyanide using ion exchange chromatography in Neisseria meningitidis serogroups A, C, W, Y and X conjugates used in vaccine manufacture.
Polysaccharide vaccines essentially used in the prevention of bacterial infections are known to be good immunogens when conjugated to an immunogenic protein using various cyanylating agents. Analysis of residual cyanide in polysaccharide conjugate vaccines is an ardent task due to the complexity of the sample matrices and the lack of suitable methods. We report a selective ion chromatography method with electrochemical detection using IonPac AS7 column for estimation of residual cyanide in meningococcal serogroups A, C, W, Y and X bulk conjugates in presence of other interfering ions. Gold electrode and Ag/AgCl reference electrode ensures sensitivity and reproducibility of cyanide quantitation. The calibration curve of the method is linear having r2 ≥0.990 over the concentration range 1.45 ng/mL to 93.10 ng/mL. The recovery of cyanide in bulk conjugates ranged between 96.0% and 108.9%. The limits of detection and quantitation were 0.50 ng/mL and 1.45 ng/mL which corresponds to 0.31 ng/μg and 0.91 ng/μg of polysaccharide respectively. The method validation and feasibility study were performed using Men W and Men X bulk conjugates respectively with in house residual cyanide specification due to unavailability of pharmacopeia guidelines. The method is reproducible and can accurately quantify residual cyanide in purified meningococcal bulk conjugates.


Mathematical Basis of Predicting Dominant Function in Protein Sequences by a Generic HMM-ANN Algorithm.
The accurate annotation of an unknown protein sequence depends on extant data of template sequences. This could be empirical or sets of reference sequences, and provides an exhaustive pool of probable functions. Individual methods of predicting dominant function possess shortcomings such as varying degrees of inter-sequence redundancy, arbitrary domain inclusion thresholds, heterogeneous parameterization protocols, and ill-conditioned input channels. Here, I present a rigorous theoretical derivation of various steps of a generic algorithm that integrates and utilizes several statistical methods to predict the dominant function in unknown protein sequences. The accompanying mathematical proofs, interval definitions, analysis, and numerical computations presented are meant to offer insights not only into the specificity and accuracy of predictions, but also provide details of the operatic mechanisms involved in the integration and its ensuing rigor. The algorithm uses numerically modified raw hidden markov model scores of well defined sets of training sequences and clusters them on the basis of known function. The results are then fed into an artificial neural network, the predictions of which can be refined using the available data. This pipeline is trained recursively and can be used to discern the dominant principal function, and thereby, annotate an unknown protein sequence. Whilst, the approach is complex, the specificity of the final predictions can benefit laboratory workers design their experiments with greater confidence.


Isothiocyanates stimulating oviposition by the diamondback moth, Plutella xylostella.
Recognition of cabbage as a host plant for the diamondback moth (DBM) has previously been shown to depend on compounds that are extracted by soaking intact foliage in chloroform. Analysis of such chloroform extracts by open column chromatography has now resulted in the isolation of highly active fractions that elicit oviposition on treated filter papers. Further separation of these fractions by high-performance liquid chromatography revealed the presence of two distinct groups of active compounds that may be classified as volatile and non-volatile. The two prominent volatile components were separated and identified by mass spectrometry as the isothiocyanates, iberin (3-methylsulfinylpropyl isothiocyanate) and sulforaphane (4-methylsulfinyl-3-butenyl isothiocyanate). Subsequent bioassays of a range of isothiocyanates showed that iberin and sulforaphane were the most active of those tested. Other isothiocyanates with sulfur in the side chain were also active, whereas alkyl and phenyl isothiocyanates had only limited activity. In electrophysiological experiments, electroantennograms (EAGs) indicated positive responses of moth antennae to the isothiocyanates that were most active in behavioral assays. Since sulforaphane has been identified as a major inducer of anticarcinogenic activity in mouse tissue, a synthetic analog (exo-2-acetyl-5-isothiocyanatonorbornane) that shows similar inducer activity was tested on DBM. This bicyclic analog was highly active in both behavioral and EAG assays, suggesting similarity in receptor sites for the two types of biological activity.


Chromatographic determination of N-acetyl-DL-tryptophan and octanoic acid in human albumin solutions.
Chromatographic procedures have been developed for determination of the stabilizers N-acetyl-DL-tryptophan and octanoic acid in human albumin solutions. N-Acetyl-DL-tryptophan and the internal standard, N-formyl-DL-tryptophan, were separated by liquid chromatography on a reversed-phase column with UV detection at 280 nm. Deproteinization and extraction were carried out with methanol. The extraction recovery at the level of 4.9 mM was 92.5 +/- 2.5% (S.D.) (n = 10), and the average coefficient of variation (C.V.) for replicate analyses of albumin solutions (mean = 2.57, 10.44 and 17.10 mM) was 1.10% (n = 27). Octanoic acid was determined gas chromatographically as its methyl ester, with nonanoic acid as the internal standard. The sample pretreatment included acidification, extraction with hexane and derivatization with methanol-sulphuric acid. The relative recovery from albumin solutions was 89.7 +/- 5.8% (S.D.) (n = 6), and replicate determinations of the compound yielded a C.V. of 5.5% (mean = 14.82 mM, n = 9).


Quantitative Morphometric Analysis of Anatomical Landmarks Involved in Bilateral Sagittal Split Osteotomy in Different Skeletal Malocclusions Using Cone-Beam Computed Tomography.
To assess and compare the anatomical landmarks involved in bilateral sagittal split osteotomy (BSSO) in different skeletal malocclusions quantitatively using cone-beam computed tomography (CBCT). This retrospective study evaluated 63 CBCT scans of the three malocclusion classes (n = 21). Twenty-one quantitative variables were measured and compared between different malocclusion classes. One-way ANOVA, Welch test, Kruskal Wallis test and the Mann-Whitney test with Bonferroni adjustment were used for data analysis. The level of significance was set at ≤0.05. The patients had a mean age of 26.35 years (range 18-58 years). The width of the lateral cortex in the posterior third (T1-P) of ramus in the left side, and the mean value in the right and left sides were significantly different between different classes of malocclusion (P = 0.049) such that class II patients had significantly thinner lateral cortex. No other significant differences were noted between different malocclusion classes in other variables. The quantitative morphological assessment of the mandible in BSSO of different malocclusion classes did not reveal significant differences. There is no superiority in the dimensions of discussed landmarks involved in BSSO among different malocclusion classes. So the risk or the safety of this kind of surgery is similar for these classes. Thus, diagnostic preoperative workup by CBCT is imperative for selection of a surgical approach with minimal postoperative complications.


Evaluation of rhizoremediation and methane emission in diesel-contaminated soil cultivated with tall fescue (Festuca arundinacea).
Rhizoremediation, CH4 emission, and bacterial community dynamics were evaluated in diesel-contaminated soil cultivated with tall fescue via a pot experiment. At the beginning of the experiment, total petroleum hydrocarbons (TPHs) removal efficiency was 30.2% in tall fescue-cultivated soil, which was significantly higher than that of unplanted soil (19.4%). However, when compost was added as a soil amendment, TPHs removal efficiency increased to 39.2% in tall fescue-cultivated soil. Interestingly, potential CH4 emissions were more affected by the initial diesel concentration than by compost addition or tall fescue planting. Specifically, the potential CH4 emission was approximately 3.8 times higher in the treatment with the highest initial diesel concentration (T-WC38) than that of the treatment with the lowest initial diesel concentration (T-WC5). Functional gene analysis revealed that TPHs removal had a linear correlation with the alkB/16S gene ratio, whereas potential CH4 emission had a linear correlation with pmoA gene copy numbers. Initial diesel concentrations in soil also affected bacterial community structures and the genera Rhizobium, Halothiobacillus, and Geobacter were found to be positively linked to diesel-contaminated soil rhizoremediation. Therefore, this study provides useful insights into the development of strategies to enhance rhizoremediation efficiency and CH4 emission mitigation in diesel-contaminated soils.


Actirhodobacter atriluteus gen. nov., sp. nov., isolated from the surface water of the Yellow Sea.
A Gram-stain-negative, aerobic, orange-pigmented bacterial strain, designated HHU K3-1 T, was isolated from the surface water of the Yellow Sea. The strain was observed to grow on 2216E agar medium, and growth occurred at pH 6.0-8.0 (optimum 7.0), 28-37 °C (optimum 28 °C), and in the presence of 0.5-5% (w/v) NaCl (optimum 1-3%). The major fatty acids (> 10%) were summed feature 3 (C16:1ω6c/C16:1ω7c), C17:1ω6c and summed feature 8 (C18:1ω6c/C18:1ω7c). Strain HHU K3-1 T was found to contain ubiquinone-10 as the predominant quinone and the major polar lipids were diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and sphingoglycolipid (SGL). The 16S rRNA gene sequence analysis indicated that strain HHU K3-1 T shared highest similarities with Pelagerythrobacter marensis KCTC 22370 T (97.7%) and Qipengyuania oceanensis MCCC 1A09965T (96.9%). However, a phylogenetic tree based on 288 orthologous clusters (OCs) indicated that HHU K3-1 T was close related to Parapontixanthobacter aurantiacus MCCC 1A09962T. The pairwise AAI and evolutionary distance between HHU K3-1 T and Parapontixanthobacter aurantiacus MCCC 1A09962T are 67.1% and 0.43, respectively, which meet the recently proposed standard to differentiate genera in the family Erythrobacteraceae. On the basis of the result obtained by the polyphasic taxonomic study, strain HHU K3-1 T can be considered to represent a novel genus in the family Erythrobacteraceae, for which the name Actirhodobacter atriluteus gen. nov., sp. nov. is proposed. The type strain is HHU K3-1 T (= MCCC 1K04225T = KCTC 72834 T = CGMCC 1.17395 T).


Mapping pollution exposure and chemistry during an extreme air quality event (the 2018 Kīlauea eruption) using a low-cost sensor network.
Extreme air quality episodes represent a major threat to human health worldwide but are highly dynamic and exceedingly challenging to monitor. The 2018 Kīlauea Lower East Rift Zone eruption (May to August 2018) blanketed much of Hawai'i Island in "vog" (volcanic smog), a mixture of primary volcanic sulfur dioxide (SO2) gas and secondary particulate matter (PM). This episode was captured by several monitoring platforms, including a low-cost sensor (LCS) network consisting of 30 nodes designed and deployed specifically to monitor PM and SO2 during the event. Downwind of the eruption, network stations measured peak hourly PM2.5 and SO2 concentrations that exceeded 75 μg m-3 and 1,200 parts per billion (ppb), respectively. The LCS network's high spatial density enabled highly granular estimates of human exposure to both pollutants during the eruption, which was not possible using preexisting air quality measurements. Because of overlaps in population distribution and plume dynamics, a much larger proportion of the island's population was exposed to elevated levels of fine PM than to SO2 Additionally, the spatially distributed network was able to resolve the volcanic plume's chemical evolution downwind of the eruption. Measurements find a mean SO2 conversion time of ∼36 h, demonstrating the ability of distributed LCS networks to observe reaction kinetics and quantify chemical transformations of air pollutants in a real-world setting. This work also highlights the utility of LCS networks for emergency response during extreme episodes to complement existing air quality monitoring approaches.


Tumour budding and CD8+ T cells: 'attackers' and 'defenders' in rectal cancer with and without neoadjuvant chemoradiotherapy.
Tumour budding ('attacker') and CD8+ T cells ('defender') are recognised as important parameters for risk stratification in colon cancers and, combined, may have an even stronger clinical impact. Here, we determine the value of tumour budding and CD8+ in rectal cancer patients treated with/without neoadjuvant therapy. Using digital scans of all tumour slides/case, we analysed CD8+ T cell counts in two patient cohorts: 45 neoadjuvantly treated and 47 primarily surgically treated (totalling n = 543 slides) after double-staining of the surgical resection specimen for pan-cytokeratin and CD8+ . Tumour buds in hot-spots were manually counted (area = 0.785 mm2 ) and CD8+ T cell counts were analysed separately both in tumour budding hot-spots and the densest CD8+ regions throughout the tumour. In neoadjuvantly treated patients, only tumour budding and not CD8+ T cells was associated with tumour features, including more advanced ypT (P = 0.0062), venous invasion (P = 0.002), lymphatic invasion (P = 0.0003) and perineural invasion (P = 0.0017), as well as higher American Joint Committee on Cancer (AJCC) tumour regression score (P = 0.0035), indicating less tumour response. Overall survival was also worse in patients with high-grade budding in univariate analysis only. In contrast, all three variables, namely tumour budding (P = 0.0347), CD8+ T cells in budding hot-spots (P = 0.0382) and CD8+ T cells in the densest areas (P = 0.0117) were also associated with worse (budding) and better (CD8) survival time in the multivariate setting. In rectal cancer, tumour budding has clinical relevance in both primarily surgically treated patients and in those with neoadjuvantly treated patients, where it characterises highly aggressive residual disease. CD8+ T cell counts appear not to have prognostic relevance in the neoadjuvant context.


Antimalarial drugs and human neutrophil oxidative metabolism.
The effect of several commonly used antimalarial drugs on human peripheral blood neutrophil oxidative metabolism was studied. The following drugs were tested: chloroquine diphosphate, quinine HCl, mefloquine, proguanil HCl, cycloguanil, pyrimethamine, sulphadoxine, and tetracycline HCl. It was found that none of the antimalarial drugs examined, at clinically obtainable concentrations, had any inhibitory effect on neutrophil oxygen consumption, glucose oxidation, superoxide production, NBT reduction, and chemiluminescence. However, at higher concentrations chloroquine, quinine, mefloquine, and proguanil inhibited neutrophil oxidative burst. There was a slight enhancing effect on neutrophil oxidative metabolism by pyrimethamine, combination of pyrimethamine-sulphadoxine, cycloguanil and tetracycline at concentrations lower than the clinical levels.


Identification of mitochondrial respiratory chain signature for predicting prognosis and immunotherapy response in stomach adenocarcinoma.
Stomach adenocarcinoma (STAD) is the third leading cause of cancer-related deaths and the fifth most prevalent malignancy worldwide. Mitochondrial respiratory chain complexes play a crucial role in STAD pathogenesis. However, how mitochondrial respiratory chain complex genes (MRCCGs) affect the prognosis and tumor microenvironment in STAD remains unclear. In this study, we systematically analyzed genetic alterations and copy number variations of different expression densities of MRCCGs, based on 806 samples from two independent STAD cohorts. Then we employed the unsupervised clustering method to classify the samples into three expression patterns based on the prognostic MRCCG expressions, and found that they were involved in different biological pathways and correlated with the clinicopathological characteristics, immune cell infiltration, and prognosis of STAD. Subsequently, we conducted a univariate Cox regression analysis to identify the prognostic value of 1175 subtype-related differentially expressed genes (DEGs) and screened out 555 prognostic-related genes. Principal component analysis was performed and developed the MG score system to quantify MRCCG patterns of STAD. The prognostic significance of MG Score was validated in three cohorts. The low MG score group, characterized by increased microsatellite instability-high (MSI-H), tumor mutation burden (TMB), PD-L1 expression, had a better prognosis. Interestingly, we demonstrated MRCCG patterns score could predict the sensitivity to ferroptosis inducing therapy. Our comprehensive analysis of MRCCGs in STAD demonstrated their potential roles in the tumor-immune-stromal microenvironment, clinicopathological features, and prognosis. Our findings highlight that MRCCGs may provide a new understanding of immunotherapy strategies for gastric cancer and provide a new perspective on the development of personalized immune therapeutic strategies for patients with STAD.


Bone reactions to longstanding functional load at implants: an experimental study in dogs.
The aims of the present investigation were (i) to study marginal bone level alterations following implant installation, abutment connection and functional loading and (ii) to analyse bone tissue reactions to functional load. Six beagle dogs, about 1-year old, were used. All mandibular pre-molars were extracted. Three months later four implants of the Astra Tech Implants Dental System were installed in one side of the mandible and four standard fixtures of the Brånemark System were placed in the contralateral side of the mandible. Abutment connection was performed 3 months later and a plaque control programme was initiated. Three months after abutment connection fixed partial dentures (FPDs) made in gold were cemented to the maxillary canines and pre-molars. FPDs were also connected to the three posterior implants in each side of the mandible, while the mesial implant in each side was used as an unloaded control. Radiographs were obtained from all implant sites following implant installation, abutment connection and FPD placement. Ten months after the FPD placement the radiographic examination was repeated. The animals were sacrificed and biopsies from all implant sites were obtained and prepared for histological analysis. The radiographic analysis revealed that largest amount of bone loss occurred following implant installation and abutment connection and that this loss was more pronounced at Brånemark than at Astra implants. The bone level alterations that were observed at implants exposed to 10 months of functional load in both implant systems were small and did not differ from control sites. The histological analysis revealed that implants exposed to functional load exhibited a higher degree of bone-to-implant contact than control implants in both implant systems. It is suggested that functional load at implants may enhance osseointegration and does not result in marginal bone loss.


Simple, all-optical, noncontact, depth-selective, narrowband surface acoustic wave measurement system for evaluating the Rayleigh velocity of small samples or areas.
In this paper a new ultrasonic testing system is described that utilizes noncontact optical methods to generate and detect surface acoustic waves (SAWs) and has significant applications in the nondestructive evaluation of surface material. A narrowband SAW is generated with a new and straightforward grating mask image projection method that provides fast switching and a controllable frequency band, and hence control of the penetration depth of the ultrasonic wave. A narrowband SAW with center frequency above 30 MHz, and hence better depth resolution, is generated. The detection of the SAW is performed with a simplified design of an optical fiber interferometer that has good sensitivity and manoeuvrability without requiring additional auxiliary components. The novel combination of these two optical techniques permits the measurement of small samples that are otherwise difficult to measure, especially nondestructively. A model was constructed to simulate the temporal characteristics of the generated narrowband SAW and showed good agreement with experiment. Measurements on an aluminum sample and an extracted human incisor demonstrate the system's performance.


Enzymatic production of N-acetyl-d-glucosamine from crayfish shell wastes pretreated via high pressure homogenization.
This study presents an efficient pretreatment of crayfish shell using high pressure homogenization that enables N-acetyl-d-glucosamine (GlcNAc) production by chitinase. Firstly, the chitinase from Serratia proteamaculans NJ303 was screened for its ability to degrade crayfish shell and produce GlcNAc as the sole product. Secondly, high pressure homogenization, which caused the crayfish shell to adopt a fluffy netted structure that was characterized by Scanning electron microscope (SEM), Fourier transform infrared spectrometer (FT-IR), X-ray diffraction (XRD), was evaluated as the best pretreatment method. In addition, the optimal conditions of high pressure homogenization of crayfish shell were determined to be five cycles at a pressure of 400bar, which achieved a yield of 3.9g/L of GlcNAc from 25g/L of crayfish shell in a batch enzymatic reaction over 1.5h. The results showed high pressure homogenization might be an efficient method for direct utilization of crayfish shell for enzymatic production of GlcNAc.


Recommendations for mental health care of older adults: Part 2--an overview of dementia, delirium, and substance abuse.
Expansion and diversification of the older adult demographic group is a key indication that nurses need to be aware of current recommended practice related to the most common mental health diagnoses in this age group. Such conditions include depression, anxiety, dementia, delirium, and substance abuse/dependence. This article, the second in a two-part series, focuses on key assessment components and suggested interventions for dementia, delirium, and substance abuse/dependence.


High glucose mediates apoptosis and osteogenesis of MSCs via downregulation of AKT-Sirt1-TWIST.
Mesenchymal stem cells have been widely used in the treatment of diabetes mellitus. However, hyperglycemia associated with DM promotes cell apoptosis and affects osteogenic differentiation of MSCs in varying degrees, leading to osteoporosis in DM patients. Therefore, in this paper, the effect of high glucose on apoptosis and osteogenesis of MSCs was investigated and underlying mechanism was further determined. Intracellular ROS levels were determined using probe DCFH-DA. MMP was detected using JC-1 staining. Cell apoptosis was detected using Annexin V-FITC/PI and Flow Cytometer. The expression of genes and protein was detected by qRT-PCR and Western blot respectively. The results showed high glucose induced MSC apoptosis but promoted its osteogenesis. Western blot analysis revealed that high glucose downregulated AKT-Sirt1-TWIST pathway. Activation of Sirt1 via SRT1720 increased TWIST expression, alleviated MSC apoptosis and promoted osteogenesis of MSCs. TWIST knockdown studies demonstrated that inhibition of TWIST intensified high glucose-induced apoptosis but promoted osteogenesis differentiation of MSCs. TWIST is likely to be a new regulator for cross talk between Sirt1 and its downstream targets. Our data demonstrates that high glucose induces MSC apoptosis and enhances osteogenesis differentiation via downregulation of AKT-Sirt1-TWIST.


Significance of estimated glomerular filtration rate in predicting brain or heart attacks in obese and non-obese populations.
The Japanese Specific Health Checkup mainly focuses on metabolic syndrome for preventing cardiovascular events. Subjects are stratified by measuring waist circumference, body mass index, blood pressure, triglycerides, and fasting plasma glucose. However, estimated glomerular filtration rate (eGFR) is not considered essential. A longitudinal cohort study assessed the association of eGFR with new-onset brain or heart attacks in a large Japanese nationwide Specific Health Checkup database. A total of 109,349 Japanese subjects (mean age 63.2 years, 39.5 % men) were examined for the events 2 years later. The odds ratios were calculated for new events in the total and subgroup populations divided by BMI < or ≥25 kg/m(2), obese and non-obese, respectively. Obese subjects were more often male and had proteinuria (dipstick test ≥1+), lower eGFR, and higher systolic and diastolic BP, fasting plasma glucose, hemoglobin A1c, and triglycerides (TG). Rates of new-onset brain or heart attacks were 3.1 and 4.0 % in the groups of non-obese and obese subjects, respectively. In the total population, eGFR as well as higher BMI (≥25 kg/m(2)), higher BP (high-normal hypertension or greater), higher TG (≥150 mg/dl), and proteinuria were significant risk factors for developing brain or heart attacks. The eGFR was significant in non-obese subjects, but not in the obese. As the ultimate aim of 'Specific Health Checkup' is to prevent cardiovascular events, our study suggests that eGFR should be evaluated in non-obese subjects.


Intensive care unit versus high dependency care unit admission after emergency surgery: a nationwide in-patient registry study.
The appropriate level of postoperative critical care for patients undergoing emergency surgery is unknown. We aimed to assess the outcomes of postoperative patients treated in the intensive care unit (ICU) and high dependency care unit (HDU) after emergency surgery. Analysis of national in-patient registry data in Japan from July 2010 to March 2018, including patients undergoing one of 10 emergency surgeries on the day of hospital admission. The exposures were ICU or HDU admission on the day of surgery. The primary outcome was in-hospital mortality. We performed multivariable logistic regression analysis adjusted for patient and hospital characteristics. Results are presented as n (%) and odds ratios (ORs) with 95% confidence intervals. We included 158 149 patients from 646 hospitals. Crude in-hospital mortality for each procedure ranged from 168/8583 (2.0%) for cholecystectomy to 2842/12 958 (21.9%) for patients undergoing surgery for traumatic brain injury. Compared with HDU admission, ICU admission was associated with lower in-hospital mortality among the cohorts for medium-mortality risk procedures (procedure-specific mortality 5-15%) (ICU: 8834/73 616 [12.0%] vs HDU: 2586/25 262 [10.2%]; OR=0.90 [0.85-0.96]; P=0.001), and high-mortality risk procedures (procedures-specific mortality >15%) (ICU: 3445/16 334 [21.1%] vs HDU: 996/4613 [21.6%]; OR=0.86 [0.78-0.96]; P=0.005). There were no differences in mortality for low-mortality risk procedures with procedure-specific mortality <5%. In this national registry study, postoperative critical care in ICU was associated with lower in-hospital mortality than in HDU for patients undergoing medium-risk and high-risk emergency surgery. Further research is needed to understand the role of critical care for surgical patients.


Determinants of cesarean delivery: a classification tree analysis.
Cesarean delivery (CD) rates are rising in many parts of the world. To define strategies to reduce them, it is important to identify their clinical and organizational determinants. The objective of this cross-sectional study is to identify sub-types of women at higher risk of CD using demographic, clinical and organizational variables. All hospital discharge records of women who delivered between 2005 and mid-2010 in the Emilia-Romagna Region of Italy were retrieved and linked with birth certificates. Sociodemographic and clinical information was retrieved from the two data sources. Organizational variables included activity volume (number of births per year), hospital type, and hour and day of delivery. A classification tree analysis was used to identify the variables and the combinations of variables that best discriminated cesarean from vaginal delivery. The classification tree analysis indicated that the most important variables discriminating the sub-groups of women at different risk of cesarean section were: previous cesarean, mal-position/mal-presentation, fetal distress, and abruptio placentae or placenta previa or ante-partum hemorrhage. These variables account for more than 60% of all cesarean deliveries. A sensitivity analysis identified multiparity and fetal weight as additional discriminatory variables. Clinical variables are important predictors of CD. To reduce the CD rate, audit activities should examine in more detail the clinical conditions for which the need of CD is questionable or inappropriate.


Drug recirculation model with multiple cycles occurring at unequal time intervals.
A pharmacokinetic model for enterohepatic recycling has been developed to take into account multiple recirculations likely to occur at various times after intravenous or subcutaneous injection, after infusion, or after a single oral administration of a drug. The times when the gall bladder empties, the duration of infusion and the number of recirculations may be arbitrarily chosen (for simulations) or computed (for optimization) to express the concentration in the central compartment at any time. Without a new theoretical calculation, the area under the concentration curve may be obtained as a function of the model parameters. As an example, the model is applied to an experimental case of four recirculations after oral administration and to a new drug data fitting.


Magnetic circular dichroism analysis of the IHP effect on spin equilibria in human ferric hemoglobins.
Magnetic circular dichroism (MCD) spectroscopy has been to explore the connection between optical spectra and the high spin population of several hemoglobins under various conditions. It is found that the effectiveness of IHP in inducing spectral changes can be markedly affected by solvent. For example, the IHP-induced spectral changes in the visible region for nitritomethemoglobin-A in mixed buffer solvent systems (glycerol or polyethylene glycol (PEG), mw 190-210) are more than double those observed in aqueous buffers. We estimate that IHP induces a mix of R/T forms in bis-tris phosphate buffers, for NO-2metHb that is only about 50%. While PEG and glycerol both lead to enhanced IHP-induced spectral differences, they behave differently in two aspects. PEG shifts the visible MCD and absorption spectra of F-metHb-A, supposedly already biased towards the T form by ligand, in the same direction that IHP does. PEG also maximizes the spin state changes with IHP for three R from hemoglobins and N-3metHb-A, and so appears to stabilize the T form in all cases. Glycerol does not. In addition, the apparent binding constant for NO-2 to H2OmetHb-A differs between these two solvents. Comparison of the data from several hemoglobins leads to the conclusion that the changes in spin state distributions induced by IHP correlate well with quaternary structure for a given hemoglobin. An analogous correlation amongst various proteins between initial spin state distribution (IHP) absent) and quaternary structure is not found.


Mucosal-associated invariant T cell responses differ by sex in COVID-19.
Sexual dimorphisms in immune responses contribute to coronavirus disease 2019 (COVID-19) outcomes, but the mechanisms governing this disparity remain incompletely understood. We carried out sex-balanced sampling of peripheral blood mononuclear cells from hospitalized and non-hospitalized individuals with confirmed COVID-19, uninfected close contacts, and healthy control individuals for 36-color flow cytometry and single-cell RNA sequencing. Our results revealed a pronounced reduction of circulating mucosal-associated invariant T (MAIT) cells in infected females. Integration of published COVID-19 airway tissue datasets suggests that this reduction represented a major wave of MAIT cell extravasation during early infection in females. Moreover, MAIT cells from females possessed an immunologically active gene signature, whereas cells from males were pro-apoptotic. Our findings uncover a female-specific protective MAIT cell profile, potentially shedding light on reduced COVID-19 susceptibility in females. This work was supported by NIH/NIAID (U01AI066569 and UM1AI104681), the Defense Advanced Projects Agency (DARPA; N66001-09-C-2082 and HR0011-17-2-0069), the Veterans Affairs Health System, and Virology Quality Assurance (VQA; 75N93019C00015). The content is solely the responsibility of the authors and does not necessarily represent the official view of the National Institutes of Health. COVID-19 samples were processed under Biosafety level 2 (BSL-2) with aerosol management enhancement or BSL-3 in the Duke Regional Biocontainment Laboratory, which received partial support for construction from NIH/NIAID (UC6AI058607).


[Autoradiographic study of the early stages of human female meiosis in organ culture].
Incorporation of H3-thymidin in developing germinative cells and the movement of the label in successive stages of preprophase and prophase of meiosis were studied in the organic culture of the ovaries of 7-12 and 23 week human embryos by the method of autoradiography. H3-thymidin was incorporated by oognia and oocytes of the last stages of preleptotene with a thin uneven network of chromatin in the nucleus and by oocytes of the stage of leptotene and, rarely, zygotene. In oocytes with prochromosomes the label appeared within 12 hours. The cells labeled in the premeiotic DNA synthesis were changed into the zygotene stage within 4 days, into the pachytene stage- within 14 days. The development took 7-14 days from the last premitosis S-period to zygotene. Oogoniums divided two times for 7 days. Gz-period of oognia -1/2 of mitosis lasts an average of 10 hours. The S-period of oogonia lasts for about 9 hours.


Estimation of divergence time between two sibling species of the Anopheles (Kerteszia) cruzii complex using a multilocus approach.
Anopheles cruzii is the primary human Plasmodium vector in southern and southeastern Brazil. The distribution of this mosquito follows the coast of the Brazilian Atlantic Forest. Previous studies indicated that An. cruzii is a complex of cryptic species. A multilocus approach using six loci, three circadian clock genes and three encoding ribosomal proteins, was implemented to investigate in more detail the genetic differentiation between the An. cruzii populations from Santa Catarina (southern Brazil) and Bahia States (northeastern Brazil) that represent two sibling species. The analysis revealed very high FST values and fixed differences between the two An. cruzii sibling species in all loci, irrespective of their function. An Isolation with Migration model was fit to the data using the IM program. The results reveal no migration in either direction and allowed a rough estimate of the divergence time between the two sibling species. Population genetics analysis of An. cruzii samples from two Brazilian localities using a multilocus approach confirmed that they represent two different sibling species in this complex. The results suggest that the two species have not exchanged migrants since their separation and that they possibly diverged between 1.1 and 3.6 million years ago, a period of intense climatic changes.


Spectroscopic and theoretical evidences for the surface binding of voglibose drug with DNA.
Binding of voglibose (VOG), an alpha glucosidase inhibitor, with CT-DNA has been investigated using various spectroscopic techniques including UV-Vis, fluorescence and circular dichroism (CD) coupled with relative viscosity. Isothermal titration calorimetric studies have been used to calculate the thermodynamic parameters such as ΔH (0.0188 cal/mol), ΔS (63.3 cal/mol/K) and ΔG (-18.8 kcal/mol), which reveal that the binding is a spontaneous process and hydrophobic and H-bonding interactions play major roles in the binding process. Effect of ionic strength confirms the existence of hydrophobic interaction between VOG and CT-DNA. Competitive displacement assays with ethidium bromide (EB) and Hoechst 33258 suggest that VOG possibly binds on the surface of CT-DNA. Viscosity measurements also disclose that the binding could be mainly surface binding. Corroborating the experimental observations, metadynamics molecular simulation studies confirm that VOG binding on the surface of the DNA molecule through hydrophobic interactions and direct and water molecule mediated H-bonding.