Programmed cell death: the battlefield between the host and alpha-herpesviruses and a potential avenue for cancer treatment.
Programed cell death is an antiviral mechanism by which the host limits viral replication and protects uninfected cells. Many viruses encode proteins resistant to programed cell death to escape the host immune defenses, which indicates that programed cell death is more favorable for the host immune defense. Alpha-herpesviruses are pathogens that widely affect the health of humans and animals in different communities worldwide. Alpha-herpesviruses can induce apoptosis, autophagy and necroptosis through different molecular mechanisms. This review concisely illustrates the different pathways of apoptosis, autophagy, and necroptosis induced by alpha-herpesviruses. These pathways influence viral infection and replication and are a potential avenue for cancer treatment. This review will increase our understanding of the role of programed cell death in the host immune defense and provides new possibilities for cancer treatment.


A conceptualisation framework for building consensus on environmental sensitivity.
Examination of the intrinsic attributes of a system that render it more or less sensitive to potential stressors provides further insight into the baseline environment. In impact assessment, sensitivity of environmental receptors can be conceptualised on the basis of their: a) quality status according to statutory indicators and associated thresholds or targets; b) statutory protection; or c) inherent risk. Where none of these considerations are pertinent, subjective value judgments can be applied to determine sensitivity. This pragmatic conceptual framework formed the basis of a stakeholder consultation process for harmonising degrees of sensitivity of a number of environmental criteria. Harmonisation was sought to facilitate their comparative and combined analysis. Overall, full or wide agreement was reached on relative sensitivity values for the large majority of the reviewed criteria. Consensus was easier to reach on some themes (e.g. biodiversity, water and cultural heritage) than others (e.g. population and soils). As anticipated, existing statutory measures shaped the outcomes but, ultimately, knowledge-based values prevailed. The agreed relative sensitivities warrant extensive consultation but the conceptual framework provides a basis for increasing stakeholder consensus and objectivity of baseline assessments. This, in turn, can contribute to improving the evidence-base for characterising the significance of potential impacts.


Paraneoplastic syndromes.
As the lengthy but nonetheless incomplete review suggests, paraneoplastic syndromes are protean in their manifestations and, for the most part, poorly understood. Indeed, some of the more common abnormalities in cancer patients that might be considered paraneoplastic--such as anorexia-cachexia syndrome or unexplained fever--have not been discussed because they are so poorly understood. Most of the syndromes reviewed are either clearly paraneoplastic or strongly associated with cancer. Their clinical importance does not lie in the number of patients affected; it is a small minority. Instead, the syndromes may occasionally be helpful in the diagnosis of cancer or in monitoring response to cancer therapy. They may also be confused with the effects of metastatic disease. In some patients, amelioration of the syndromes can reverse the patient's dominant symptoms and thus provide significant clinical palliation. In a more general context, studies of etiologic mechanisms in paraneoplastic syndromes may offer insights into a variety of unexplained abnormalities in cancer patients. The best-understood syndromes result from tumor production of biologically active substances or, to a lesser extent, from autoimmune phenomena. These would appear to be probable mechanisms in many recognized paraneoplastic syndromes of uncertain etiology and perhaps in some heretofore unrecognized paraneoplastic syndromes. Finally, paraneoplastic syndromes could also hold clues to the neoplastic process. Better understanding of the ways in which tumors regulate remote effects--such as release of TGFs--may ultimately enhance our knowledge of tumor growth itself.


Percutaneous ethanol injection for benign cystic thyroid nodules: is aspiration of ethanol-mixed fluid advantageous?
We evaluated the differences between percutaneous ethanol injection with and without aspiration of ethanol-mixed fluid for treatment of benign cystic thyroid nodules. We examined 60 patients with benign cystic thyroid nodules confirmed by fine-needle aspiration biopsy and divided them into 2 groups according to nonaspiration (group A, n = 30) or aspiration (group B, n = 30) of ethanol-mixed fluid after intracystic ethanol injection. We evaluated in both groups the complete disappearance of the cystic portion of the thyroid nodule on follow-up ultrasonography (first follow-up ultrasonography; mean, 4.6 months in group A; mean, 4.4 months in group B) (chi-square test), side effects or complications during and after the procedure (chi-square test), and the total procedure time (Student t test). Most patients showed complete disappearance of the cystic portion of the thyroid nodule (group A, n = 29; group B, n = 28), and they revealed no recurrence on follow-up ultrasonography. There was no statistical difference in the success rates between group A and group B (P > .05). Pain, the most common side effect, and other mild side effects or complications occurred in small numbers of patients in each group, but there was no significant difference in side effects or complications between the 2 groups (P > .05), except for intracystic hemorrhage (P < .05) and the complaint of all group B patients due to a double puncture (P < .001). The total procedure time was nearly double in group B than in group A because of the additional procedures, such as complete evacuation of the ethanol-mixed fluid and the 10-minute compression. Percutaneous ethanol injection without aspiration of ethanol-mixed fluid seems to be the preferable method of treatment of benign cystic thyroid nodules from the perspective of both the physician and the patient.


Serotonin neuron abnormalities in the BTBR mouse model of autism.
The inbred mouse strain BTBR T+ Itpr3tf /J (BTBR) is studied as a model of idiopathic autism because they are less social and more resistant to change than other strains. Forebrain serotonin receptors and the response to serotonin drugs are altered in BTBR mice, yet it remains unknown if serotonin neurons themselves are abnormal. In this study, we found that serotonin tissue content and the density of serotonin axons is reduced in the hippocampus of BTBR mice in comparison to C57BL/6J (C57) mice. This was accompanied by possible compensatory changes in serotonin neurons that were most pronounced in regions known to provide innervation to the hippocampus: the caudal dorsal raphe (B6) and the median raphe. These changes included increased numbers of serotonin neurons and hyperactivation of Fos expression. Metrics of serotonin neurons in the rostral 2/3 of the dorsal raphe and serotonin content of the prefrontal cortex were less impacted. Thus, serotonin neurons exhibit region-dependent abnormalities in the BTBR mouse that may contribute to their altered behavioral profile. Autism Res 2017, 10: 66-77. © 2016 International Society for Autism Research, Wiley Periodicals, Inc.


[Determination of isothiocyanates and related compounds in mustard extract and horseradish extract used as natural food additives].
Amounts of isothiocyanates and related compounds in a mustard extract and a horseradish extract for food additive use were determined by GC, after confirmation of the identity of GC peaks by GC/MS. Amounts of allyl isothiocyanate, which included that of allyl thiocyanate, because most of the allyl thiocyanate detected in the sample was assumed to have been formed from allyl isothiocyanate during GC analysis, were 97.6% and 85.4%, in the mustard extract and the horseradish extract, respectively. Total amounts of the identified isothiocyanates in the mustard extract and the horseradish extract were 98.5% and 95.4%, respectively. Allyl cyanide, a degradation product of allyl isothiocyanate, was found in the mustard extract and the horseradish extract at the levels of 0.57% and 1.73%, respectively. beta-Phenylethyl cyanide, a possible degradation product of beta-phenylethyl isothiocyanate, and allyl sulfides were found in the horseradish extract, at the levels of 0.13% and 0.46%, respectively. Allylamine, which is another degradation product of allyl isothiocyanate, was determined after acetylation, and was found in the mustard extract and the horseradish extract at the levels of 8 micrograms/g and 67 micrograms/g, respectively.


The cultural change narrative as a core component of therapeutic change.
Research indicates a similar effect of Mentalization-based treatment (MBT) and Dialectical behavior therapy (DBT) for borderline personality disorder (BPD). However, there is a paucity in studies investigating the change narrative received from and developed in these treatments. The aim of the present study is to investigate similarities and differences in the change narratives provided by MBT and DBT, and how these narratives reflect the rationale, explanations, and procedures of the provided treatment. The study is a qualitative analysis of seven interviews conducted by the authors. Three of the participants had received MBT, and four of the participants had received DBT. This study presents an Interpretative Phenomenological Analysis (IPA) of the change narratives received in two specialized treatments for BPD. The main findings from the IPA were that the change narratives described by the participants reflected the treatment they received. The DBT participants highlighted explicit learning of tools and techniques, with predictable and safe therapists. In contrast, the MBT participants emphasized a long-lasting process of exploring to create procedural learning with therapists who followed their lead. The participants' stories of change shed light on how a change narrative was developed, and therefore how the rationale, explanations and procedures were conveyed differently by MBT and DBT.


Metabolism of benzo[a]pyrene and DNA adduct formation in cultured human epidermal keratinocytes.
Cultured human epidermal cells which require no feeder layer were used to study metabolism of benzo[a]pyrene (BP) and DNA adduct formation. The cultures were prepared from a single cell suspension and maintained at a pH of 5.9--6.2. At 2 microM BP some cell toxicity was observed, and substantial cell death occurred at 4 microM BP. The metabolism and DNA binding of BP were followed from 6 to 48 h of incubation. High pressure liquid chromatography (h.p.l.c.) revealed that BP was metabolized into 9,10-diol, 7,8-diol, quinones, phenols and tetraols of BP. The DNA binding levels increased linearly up to 28 h of incubation. At 18 h, the level of DNA binding at 0.4 microM BP was 1.5 x 10(-6) mol BP/mol DNA and increased to 6.0 x 10(-6) mol BP/mol DNA at a dose of 4 microM BP. Analysis of the DNA adducts by h.p.l.c. indicates that the 2'-deoxy-N2-(7,8,9,10-tetrahydro-7 beta-,8 alpha, 9 alpha-trihydroxybenzo[a]pyrene-10-yl) guanosine was the predominant adduct formed in cells exposed to BP. The prevalence of the minor DNA adducts varies as a function of the source of the primary skin cells. These results confirm that human cells with no feeder layer metabolize BP and the resultant DNA damage is similar to that found in other mammalian systems.


Pharmaco-dynamics of human menopausal gonadotrophin (HMG) and follicle-stimulating hormone (FSH). The importance of the FSH concentration in initiating follicular growth in polycystic ovary-like disease.
Using a randomized double-blind cross-over design, the pharmaco-dynamic and pharmaco-kinetic properties of 'pure' follicle-stimulating hormone (FSH) (Metrodin) and human menopausal gonadotrophin (HMG) (Pergonal) were studied in 24 women with polycystic ovary-like disease (PCOD) during induction of ovulation. Fifty-six cycles were stimulated with FSH and 60 cycles with HMG, according to a standard protocol. Gonadotrophins were administered i.v. in a pulsatile fashion using pulse frequencies of either 30 or 120 min. The cycles stimulated with either 30 or 120 min pulse intervals showed no differences among themselves. During the stimulation phase, the FSH and HMG stimulated cycles showed equal and dose dependent FSH concentrations (mean +/- SD). The luteinizing hormone (LH) concentrations (mean +/- SD) were also equal but unchanged compared to the mean basal concentration. The LH, FSH, total urinary oestrogen excretion, and testosterone profiles (mean +/- SD) obtained from cycle days -10 to 0 as well as the pregnanediol profiles obtained from cycle days 0 to +14 showed no differences either. The occurrence of an endogenous preovulatory LH surge was significantly more frequent in the cycles stimulated with a pulse interval of 30 min compared to the cycles stimulated with a pulse interval of 120 min. The addition of LH as provided in HMG did not influence the FSH threshold concentration above which initiation of follicular growth occurred, since no differences were found in the FSH 'stable' concentrations between FSH and HMG stimulated cycles. However, intra- and inter-individual variation in the FSH 'stable' concentration at which follicular growth was initiated became obvious.(ABSTRACT TRUNCATED AT 250 WORDS)


Impact of donepezil hydrochloride on the care burden of family caregivers of patients with Alzheimer's disease.
To evaluate the impact of donepezil hydrochloride on the care burden on family members of patients with Alzheimer's disease (AD). At present, donepezil is the only drug approved for the treatment of AD in Japan. Although the care burden on primary caregivers of AD patients comprises both physical and psychological burdens and donepezil is recognized to improve cognitive dysfunction and associated symptoms, there are few data on the effects of the drug on the care burden. Of the uninstitutionalized AD patients who visited a dementia clinic between June 2008 and May 2009 with their primary family caregivers, 416 subjects who satisfied the enrollment criteria were registered for the study. All participants provided informed consent. Assessment included changes in scores on the Japanese version of the Zarit Caregiver Burden Interview (J-ZBI) and the Mini-Mental State Examination (MMSE), as well as the presence of behavioral and psychological symptoms of dementia (BPSD). Caregivers answered the questionnaires at baseline and after 12 weeks treatment with donepezil (starting dose 3 mg, p.o., once daily, followed by 5 mg after 1 or 2 weeks). There were significant changes in mean scores on the J-ZBI (-1.9 +/- 9.5; P < 0.01) and MMSE (+0.9 +/- 2.9; P < 0.01) from baseline to Week 12, without significant correlation between these two scores. In patients with BPSD, there was a significant decrease in J-ZBI scores over the 12 weeks (P = 0.013); in contrast, in patients without BPSD, the decrease in the J-ZBI score did not reach statistical significance (P = 0.418). The results indicate that donepezil improves cognitive function and some of the BPSD. As a possible consequence of improvements in BPSD, donepezil may also reduce caregivers' burden.


Improved detection of beta-lactamase activity in isolates of Staphylococcus saprophyticus with the use of a modified Cefinase disk procedure.
The Cefinase disk (BBL Microbiology Systems, Cockeysville, MD) technique has been reported to be relatively insensitive for detection of beta-lactamase production by Staphylococcus saprophyticus. The effects of time (up to 24 hours) and temperature (25 degrees C vs. 35 degrees C) of test incubation as well as enzyme induction on Cefinase detection of beta-lactamase activity in clinical isolates of S. saprophyticus was investigated. Results were compared to beta-lactamase detection with the use of conventional nitrocefin and cloverleaf methods and with penicillin minimal inhibitory concentrations (MICs). Of 85 S. saprophyticus isolates, 84 (98.8%) had penicillin MIC end points of greater than or equal to 0.25 microgram/mL. Beta-lactamase was detected by at least one method in 73 (85.9%) of the isolates: 63 (74.1%) by nitrocefin, 55 (64.7%) by cloverleaf, and 63 (74.1%) by Cefinase. Of Cefinase-positive isolates, 56 (88.9%) and 63 (100%) were detected at 25 degrees C and 35 degrees C, respectively, and none required enzyme induction. At 35 degrees C, 4 (6.3%) and 19 (30.2%) of the Cefinase-positive isolates were detected at 1 hour and 24 hours, respectively. Incubation of Cefinase tests at 35 degrees C for 24 hours provides a satisfactory alternative to the conventional nitrocefin assay for detection of beta-lactamase activity in S. saprophyticus.


Unique clinical features of cryptococcal meningitis among Chinese patients without predisposing diseases against patients with predisposing diseases.
The clinical features of cryptococcal meningitis (CM) in patients without predisposing diseases (PD) remain unclear. In sum, 162 of the 167 patients without PD and 162 of the 309 patients with PD were enrolled after propensity score matching. Demographic characteristics, symptoms, blood, and cerebrospinal fluid (CSF) characteristics were compared between the two groups. Kaplan-Meier curves and a Cox proportional hazards model were used to assess the factors associated with 10-week mortality. In total, approximately 35.1% of CM patients were without PD. CM patients without PD had blood profiles of higher white blood cells (WBC) [8.9(6.7-11.0) × 109/l], hemoglobin (128.4 ± 20.9 g/l), platelets [(226.2 ± 64.1) × 109/l], and serum albumin (41.2 ± 5.8 g/l) (all P ≤ .001) and CSF profiles of lower glucose (2.0 ± 1.2 mmol/l), pleocytosis [65.0 (18.0-160.0) × 106/l] and higher total protein [0.9 (0.7-1.4)g/l] (all P < .05). CM patients without PD had lower Cryptococcus culture positivity in CSF (62.5% vs. 74.1%, P = .039) but higher 2-week of CSF culture sterilization rates (69.4% vs. 51.3%, P = .031). The overall 10-week survival rate was 84.7% in patients without PD and 81.1% in patients with PD (Log-rank P = .439). CSF glucose <1.5 mmol/l, CSF fungal burden >20 cells/high power field and treatment lacking amphotericin B had a 3-4 times higher risk of death in patients without PD, whereas serum albumin <35 g/l, CSF glucose < 1.5 mmol/l, and CSF WBC <55 × 106 cell/l were risk factors for patients with PD. CM patients without PD had unique blood and CSF profiles, especially, had lower Cryptococcus culture positivity in CSF, and higher 2-week CSF culture sterilization. Low CSF glucose levels, higher fungal burden, and treatment without amphotericin B were risk factors for 10-week mortality.


Effects of reclaimed waters on spectral properties and leaf traits of citrus orchards.
Effects resulting from the use of reclaimed waters on mandarins and grapefruits are evaluated by measuring the spectral responses of their canopies and the anatomy and the chlorophyll content of their leaves against control trees irrigated with waters provided by an interbasin transfer. Spectral responses from the red (R) and near-infrared (NIR) wavelength bands, and its normalized ratio (NDVI), were acquired from a hyperspatial flight conducted after a low-moderate exposition to reclaimed waters. Chlorophyll and leaf and palisade/spongy ratio thicknesses were analyzed after a moderate-high exposition. Significant differences between controls and treatments were detected in mandarins in R and leaf chlorophyll, but not in grapefruits, likely because of their higher tolerance to saline waters. Reused waters did not affect either NIR-NDVI or anatomy traits. Hyperspatial sensing techniques are suitable for detecting chlorophyll dynamics, but NIR information and related vegetation indices may mask the detection of periods of saline stress in citrus orchards.


Modulating the M-M distance in dinuclear complexes. New ligand with a 2,2'-Biphenol fragment as key unit: synthesis, coordination behavior, and crystal structures of Cu(II) and Zn(II) dinuclear complexes.
The synthesis and characterization of the new polyamino-phenolic ligand 3,3'-bis[N,N-bis(2-aminoethyl)aminomethyl]-2,2'-dihydroxybiphenyl (L) are reported. L contains two diethylenetriamine units linked by a 1,1'-bis(2-phenol) group (BPH) on the central nitrogen atom which allows two separate binding amino subunits in a noncyclic ligand. The basicity and binding properties of L toward Cu(II) and Zn(II) were determined by means of potentiometric measurements in aqueous solution (298.1 +/- 0.1 K, I = 0.15 mol dm-3). L behaves as a pentaprotic base and as a monoprotic acid under the experimental conditions used, yielding the H5L5+ or H-1L- species, respectively. L forms both mono- and dinuclear species with both metal ions investigated; the dinuclear species are largely prevalent in aqueous solution with a L/M(II) molar ratio of 1:2 at pH higher than 7. L shows different behavior in Cu(II) and Zn(II) binding, affecting the dinuclear species formed and the distance between the two coordinated metal ions, which is greater in the Zn(II) than in the Cu(II) dinuclear species. This difference can be attributed to the different degree of protonation of BPH which influences the angle between the phenyl rings in the two systems. In this way, it is possible to modulate the M(II)-M(II) distance by the choice M(II) and to space the two M(II) farther away than was possible with the previously synthesized ligands. L does not saturate the coordination sphere of the coordinated M(II) ions in the dinuclear species, and thus, these latter species are prone to add guests. 1H and 13C NMR experiments carried out in aqueous solution, as well as the crystal structures of the dinuclear Cu(II) and Zn(II) species formed in aqueous solution, aided in elucidating the involvement of L and BPH in Zn(II) and Cu(II) stabilization.


Production of itaconic acid using metabolically engineered Escherichia coli.
An Escherichia coli system was engineered for the heterologous production of itaconic acid via the expression of cis-aconitate decarboxylase gene (cad), and then maximal itaconic acid levels produced by engineered E. coli were evaluated. Expression of cad in E. coli grown in Luria-Bertani (LB) medium without glucose in a test tube resulted in 0.07 g/L itaconic acid production after 78 h at 20°C. To increase itaconic acid production, E. coli recombinants were constructed by inactivating the isocitrate dehydrogenase gene (icd) and/or the isocitrate lyase gene (aceA). Expression of cad and inactivation of icd resulted in 0.35 g/L itaconic acid production after 78 h, whereas aceA inactivation had no effect on itaconic acid production. The intracellular itaconate concentration in the Δicd strain was higher than that in the cad-expressing strain without icd inactivation, which suggests that the extracellular secretion of itaconate in E. coli is the rate-determining step during itaconic acid production. pH-stat cultivation using the cad-expressing Δicd strain in LB medium with 3% glucose in a jar fermenter resulted in 1.71 g/L itaconic acid production after 97 h at 28°C. To further increase itaconic acid production, the aconitase B gene (acnB) was overexpressed in the cad-expressing Δicd strain. Simultaneous overexpression of acnB with the expression of cad in the Δicd strain led to 4.34 g/L itaconic acid production after 105 h. Our findings indicate that icd inactivation and acnB overexpression considerably enhance itaconic acid production in cad-expressing E. coli.


Neuropsychological state of the population living in the Aral Sea region (zone of ecological crisis).
Background The Aral Sea crisis has led to harmful effects on human habitat. In recent years, mild cognitive impairment is a growing problem. Objectives This article provides the results of studying the neuropsychological state of residents living in the crisis zone of the Aral Sea region in the case of Shalkar city. We have provided an assessment of the neuropsychological state of examined population and determined the leading pathology in this region. Methods The survey sample included 344 persons of reproductive age from 21 to 45 years. We have obtained results in biochemical studies, indicating perturbations of proteometabolism and lipid metabolism. Results A correlation analysis showed dependence between a decrease of albumin and high-density lipoproteins, an increase of low-density lipoproteins and parameters of cognitive function. Conclusions The research suggests a high prevalence of cerebrovascular pathology among the population, changes in cognitive function parameters, long-term and short-term memory problems and high levels of depression.


Hair Cortisol Concentrations Are Associated with Hair Growth Rate.
There is a growing interest in hair cortisol concentrations as a valuable biomarker for the assessment of metabolic diseases and chronic psychological stress. Fifty-three volunteers were recruited, and hair segments proximal to the scalp were collected from each individual. A cost-effective ball mill was used for the preparation of hair samples, and ELISA was performed to analyze cortisol concentrations. Results indicate that the frequency of hair washing affects the hair cortisol concentration. The group that washed their hair every day had significantly lower cortisol concentrations than the group that washed it less often. However, no significant differences were detected between cosmetic-treated and nontreated hair samples. The study also shows that hair cortisol concentrations in the first 3 cm of hair segments proximal to the scalp corresponded to average hair growth rate based on 1 cm/month. Thus, hair cortisol concentrations of segments 3 cm proximal to the scalp may represent cumulative stress exposure over the previous 3 months. These findings will allow more widespread research to validate the utility of hair cortisol as a potential biomarker to assess chronic stress.


Update in vitamin D and multiple sclerosis.
Multiple sclerosis (MS) is a common neurological disease, and its etiology remains unknown. In recent years, increasing attention has been paid to the possible association between MS and vitamin D deficiency. Here, we review the current literature between MS and vitamin D, showing clear evidence that vitamin D deficiency is a risk factor for MS despite the lack of direct evidence for the effects of vitamin D in MS progression.


Phaeolschidins A-E, five hispidin derivatives with antioxidant activity from the fruiting body of Phaeolus schweinitzii collected in the Tibetan Plateau.
Five new hispidin derivatives, phaeolschidins A-E (1-5), as well as two known natural products, pinillidine (6) and hispidin (7), were isolated from the fruiting bodies of Phaeolus schweinitzii collected in the Tibetan Plateau. The structures of the new compounds were elucidated by spectroscopic methods. Phaeolschidins A-D (1-4) are new bishispidins. Phaeolschidin E (5) is a new class of hispidin derivative in which one pyrrolidin-2-one moiety was linked to C-3 of hispidin. The antioxidant activity of 1-7 was evaluated using three methods: the DPPH scavenging assay, the total antioxidant capacity assay, and the lipid peroxidation assay. Hispidin showed the strongest antioxidant activity of all tested compounds. This is the first report of secondary metabolites from the fungus P. schweinitzii.


Tumor distribution of bromodeoxyuridine-labeled cells is strongly dose dependent.
Bromodeoxyuridine (BrdUrd) is used extensively to measure the fraction of proliferating cells in tumors. Unlike endogenous markers of proliferation such as proliferating cell nuclear antigen (PCNA) and Ki-67, BrdUrd is exogenously administered and reaches the tumor via vasculature where it must then diffuse throughout the tissue to label S-phase cells. In this study, we examine the dose dependence of BrdUrd on the tumor distribution of labeled cells in histological sections. Analysis of the distribution of labeled cells in SiHa tumor xenografts showed that a dose between 400 and 1000 mg/kg was required to label cells 150 micro m from blood vessels, approaching the border of necrosis. Lower doses resulted in only the cells close to blood vessels being labeled. Interestingly, cells residing furthest from blood vessels still labeled albeit at half the level of cells situated proximal to the tumor vasculature. Results were compared with the penetration of BrdUrd seen in vitro using multilayered cell culture (MCC), a three-dimensional tissue culture model of solid tumors. Using MCC, an exposure of 100 micro M BrdUrd for 1 h was required for labeling of S-phase cells 150 micro m into the tissue, whereas cells adjacent to the edge of the tissue could be adequately labeled with just 5 micro M BrdUrd for 1 h. The area under the curve for a 100 mg/kg BrdUrd dose in mice was found to be approximately 30 micro M x h.


AICRF: ancestry inference of admixed population with deep conditional random field.
Ancestry inference of admixed populations is an important issue in anthropology and studies of gene discovery, and characterization. Usually, local ancestor inference (LAI) methods use fixed-length windows to divide chromosomes into smaller blocks. The accuracy of LAI algorithms will decrease if a window with an inappropriate length is used to infer the ancestry of admixed individuals. In this study, we first present a heuristic function to determine a proper window length for LAI methods. This heuristic is based on the distance between the ancestral populations of admixed individuals. Then we introduce a method for ancestry inference of admixed population with deep conditional random field (AICRF). AICRF uses a conditional random field (CRF) parameterized by probable extreme learning machines (PELMs) trained on reference panels where PELM is a novel probabilistic ELM classifier. This method does not require many statistical or biological parameters. We evaluate the performance of AICRF in comparison with RFMix. Experimental results show that AICRF is more accurate than RFMix with increasing admixture times.


Transcript profiling of a novel plant meristem, the monocot cambium.
While monocots lack the ability to produce a vascular cambium or woody growth, some monocot lineages evolved a novel lateral meristem, the monocot cambium, which supports secondary radial growth of stems. In contrast to the vascular cambium found in woody angiosperm and gymnosperm species, the monocot cambium produces secondary vascular bundles, which have an amphivasal organization of tracheids encircling a central strand of phloem. Currently there is no information concerning the molecular genetic basis of the development or evolution of the monocot cambium. Here we report high-quality transcriptomes for monocot cambium and early derivative tissues in two monocot genera, Yucca and Cordyline. Monocot cambium transcript profiles were compared to those of vascular cambia and secondary xylem tissues of two forest tree species, Populus trichocarpa and Eucalyptus grandis. Monocot cambium transcript levels showed that there are extensive overlaps between the regulation of monocot cambia and vascular cambia. Candidate regulatory genes that vary between the monocot and vascular cambia were also identified, and included members of the KANADI and CLE families involved in polarity and cell-cell signaling, respectively. We suggest that the monocot cambium may have evolved in part through reactivation of genetic mechanisms involved in vascular cambium regulation.


Identification of the mgc1 mutation which affects mating-pheromone-induced morphogenesis in the yeast Saccharomyces cerevisiae.
Cells of the yeast Saccharomyces cerevisiae undergo morphogenesis in response to the mating pheromones a- and alpha-factors. The 'shmoo' morphology involves localized cell surface projection formation and cytoskeleton protein synthesis. This polarization is presumed to be a prelude to mating between mating partners with opposite mating types, a and alpha. To identify genes involved in pheromone-induced morphogenesis, a system in which a gpa1 fus3 double mutant was used as a parent strain was developed, and mutants which showed altered morphogenesis in response to mating pheromone were identified. The mutation was designated mgc1 for morphogenesis control by mating pheromones. The mgc1 mutant arrested cell division in response to alpha-factor and mated with cells of the opposite mating type, but did not form a typical projection in response to pheromones.


[Cerebral MRI and evoked potentials in Wilson disease. Comparison of findings in patients with neurological follow-up].
Wilson's disease is caused by toxic copper accumulation, which leads predominantly to hepatic and basal ganglia damage. Characteristic findings in MRI and electrophysiologic examinations are described according to the occurrence of neurological symptoms. In the present study, 28 patients suffering from Wilson's disease (neurological type) were investigated. The results of MRI are compared with abnormalities of evoked potentials (BAEP, MSEP, T-VEP, MEP). All patients show hypodensities in the basal ganglial area (putamen and GI. pallidus) regularly combined with atrophy of the cerebrum and cerebellum in MRI. Signal abnormalities in the mesencephalic region (46% occurrence) and Nc. dentatus (36% occurrence) are combined with the other findings in variable patterns. Only slight changes are found in the pontine region. BAEP are disturbed in 71% of all cases and MSEP in 46%. Combined abnormalities of BAEP and MSEP were found in 39%. Pathological values occurred with a lower frequency in T-VEP (36%) and MEP (39%). The comparison of MRI findings with electrophysiological data done separately for each patient reveals no strong correlation between both methods. Individual MRI findings do not correspond with the patterns of disturbed evoked potentials and vice versa. Therefore we conclude that these methods, MRI and electrophysiological evaluation, supplement each other. Magnetic resonance imaging and electrophysiological evaluation should be performed simultaneously for therapy monitoring.


Identification, evaluation, therapy, and follow-up for children with vocal nodules in a public school setting.
This paper describes procedures used by speech and otorhinolaryngology specialists in the Independent School District of Carrollton and Farmers Branch, Texas, to identify, evaluate, treat, and follow up children with voice disorders and presents some initial findings resulting from the use of these procedures. In the identification process, 34 children were found to have vocal nodules, of which 31 children received treatment and underwent appropriate follow-up. After two months of voice therapy, 21 (68%) of the children exhibited reduced nodule size and seven (23%) exhibited normal larynges. Following six months of therapy, 26 (84%) children exhibited reduced nodule size, and 20 (65%) had normal larynges.


Reflection on Molecular Approaches Influencing State-of-the-Art Bioremediation Design: Culturing to Microbial Community Fingerprinting to Omics.
Bioremediation is generally viewed as a cost effective and sustainable technology because it relies on microbes to transform pollutants into benign compounds. Advances in molecular biological analyses allow unprecedented microbial detection and are increasingly incorporated into bioremediation. Throughout history, state-of-the-art techniques have informed bioremediation strategies. However, the insights those techniques provided were not as in depth as those provided by recently developed omics tools. Advances in next generation sequencing (NGS) have now placed metagenomics and metatranscriptomics within reach of environmental engineers. As NGS costs decrease, metagenomics and metatranscriptomics have become increasingly feasible options to rapidly scan sites for specific degradative functions and identify microorganisms important in pollutant degradation. These omic techniques are capable of revolutionizing biological treatment in environmental engineering by allowing highly sensitive characterization of previously uncultured microorganisms. Omics enables the discovery of novel microorganisms for use in bioaugmentation and supports systematic optimization of biostimulation strategies. This review describes the omics journey from roots in biology and medicine to its current status in environmental engineering including potential future directions in commercial application.


Strain-tuned optoelectronic properties of hollow gallium sulphide microspheres.
Sulfide semiconductors have attracted considerable attention. The main challenge is to prepare materials with a designable morphology, a controllable band structure and optoelectronic properties. Herein, we report a facile chemical transportation reaction for the synthesis of Ga2S3 microspheres with novel hollow morphologies and partially filled volumes. Even without any extrinsic dopant, photoluminescence (PL) emission wavelength could be facilely tuned from 635 to 665 nm, depending on its intrinsic inhomogeneous strain distribution. Geometric phase analysis (GPA) based on high-resolution transmission electron microscopy (HRTEM) imaging reveals that the strain distribution and the associated PL properties can be accurately controlled by changing the growth temperature gradient, which depends on the distance between the boats used for raw material evaporation and microsphere deposition. The stacking-fault density, lattice distortion degree and strain distribution at the shell interfacial region of the Ga2S3 microspheres could be readily adjusted. Ab initio first-principles calculations confirm that the lowest conductive band (LCB) is dominated by S-3s and Ga-4p states, which shift to the low-energy band as a result of the introduction of tensile strain, well in accordance with the observed PL evolution. Therefore, based on our strain driving strategy, novel guidelines toward the reasonable design of sulfide semiconductors with tunable photoluminescence properties are proposed.


Absence of beta-exotoxin in Thuricide preparations.
The biological insecticide Thuricide is produced from B. thuringiensis, Berliner, var. kurstaki (serotype 3a, 3b), a bacterial strain which does not synthesize exotoxin. Thus, our product is devoid of any C-mitotic or mutagenic potentiality such as is to be found in exotoxin.


Autism Spectrum Disorder in Children with an Early History of Paediatric Acquired Brain Injury.
Autism spectrum disorder (ASD) is a neurodevelopmental condition that arises from a combination of both genetic and environmental risk factors. There is a lack of research investigating whether early acquired brain injury (ABI) may be a risk factor for ASD. The current study comprehensively reviewed all hospital records at The Brain Injury Service, Kids Rehab at the Children's Hospital at Westmead (Australia) from January 2000 to January 2020. Of the approximately 528 cases, 14 children with paediatric ABI were subsequently given an ASD diagnosis (2.7%). For this ASD sample, the mean age at the time of the ABI was 1.55 years, indicating a high prevalence of early ABI in this diagnostic group. The mean age of ASD diagnosis was, on average, 5 years later than the average ASD diagnosis in the general population. Furthermore, 100% of children had at least one medical comorbidity and 73% had three or more co-occurring DSM-5 diagnoses. Although based on a small data set, results highlight early paediatric ABI as a potential risk factor for ASD and the potential for a delayed ASD diagnosis following early ABI, with comorbidities possibly masking symptoms. This study was limited by its exploratory case series design and small sample size. Nonetheless, this study highlights the need for longitudinal investigation into the efficacy of early screening for ASD symptomatology in children who have sustained an early ABI to maximise potential intervention.


Enhancing antioxidant availability in wheat grains from plants grown under seawater stress in response to microalgae extract treatments.
The present study was designed to investigate the enhanced antioxidant capacity of whole grains from wheat plants grown under seawater stress in response to microalgae extract treatment. The total carotenoid (TCAR), tocopherol (TOC), phenolic (TPC), and protein (PC) contents in whole grains of wheat plants irrigated with 10% and 20% (v/v) seawater (SW) in response to water extracts of microalgae Spirulina maxima (SME) and Chlorella ellipsoida (CEE) as well as exogenous plant growth enhancers of ascorbic acid and benzyladenine treatments were measured. The results showed that the levels of enhanced TCAR (range 0.08-0.14 g kg(-1)), TOC (range 0.05-0.12 g kg(-1)), TPC (range 0.80-2.96 g kg(-1)) and PC (range 93.4-137.9 g kg(-1)) in wheat grains of plants irrigated with 10% and 20% SW were significantly increased in response to SME and CEE treatments. Evaluation of antioxidant activity of ethanolic extracts of grains of SW-stressed plants indicated that DPPH and TBAS radical scavenging activity was significantly increased in response to SME and CEE treatments and coincided with the increase in levels of antioxidant compounds present in each extract. The electrophoretic profiles of the grains of proteins of treated samples exhibited quite different patterns from those in control samples. The results suggest that the application of algal extracts to wheat plants irrigated with SW is useful for improvement of salinity tolerance. This effect can be triggered by the stimulation of antioxidant components and protein content.


A High-Resolution Spatial Model to Predict Exposure to Pharmaceuticals in European Surface Waters: ePiE.
Environmental risk assessment of pharmaceuticals requires the determination of their environmental exposure concentrations. Existing exposure modeling approaches are often computationally demanding, require extensive data collection and processing efforts, have a limited spatial resolution, and have undergone limited evaluation against monitoring data. Here, we present ePiE (exposure to Pharmaceuticals in the Environment), a spatially explicit model calculating concentrations of active pharmaceutical ingredients (APIs) in surface waters across Europe at ∼1 km resolution. ePiE strikes a balance between generating data on exposure at high spatial resolution while having limited computational and data requirements. Comparison of model predictions with measured concentrations of a diverse set of 35 APIs in the river Ouse (UK) and Rhine basins (North West Europe), showed around 95% were within an order of magnitude. Improved predictions were obtained for the river Ouse basin (95% within a factor of 6; 55% within a factor of 2), where reliable consumption data were available and the monitoring study design was coherent with the model outputs. Application of ePiE in a prioritisation exercise for the Ouse basin identified metformin, gabapentin, and acetaminophen as priority when based on predicted exposure concentrations. After incorporation of toxic potency, this changed to desvenlafaxine, loratadine, and hydrocodone.


[The choice of antibiotic according the results of standard bacteriological analysis and express-method without isolation of pure cultures.]
The efficiency of choosing antibiotics using two methods was analyzed. The first method is based on standard laboratory analysis including isolation of pure cultures of prospective agent and detection of their individual sensitivity to antibiotics. The second method is based on a new express-technique permitting to simultaneously evaluate effect of pharmaceuticals on cultivated and not cultivated yet bacteria without isolation of pure cultures. At that, the second method permits to determine efficiency of antibiotics namely in those concentrations that can be made in the given nidus of infection. The study demonstrated existence of discrepancy between results obtained by using the given methods. The cause of discrepancy can be presence of not cultivated yet bacteria in pathologic sample. In favor of this assumption testify the received data concerning samples from patients where were presented a significant number of spore-former bacteria having additional mechanisms of resistance to antibiotics. Furthermore, the implemented study established antibiotics characterized by the greatest discrepancy of results obtained by applied methods that indicates the necessity of revision of schemes of implementation of start or empiric therapy.


Activation and immunoregulation of antigen-specific human b lymphocyte responses: multifaceted role of the monocyte.
The multifaceted role of the monocyte in the induction and modulation of antigen-specific antibody responses by human B cells was delineated. Monocytes were absolutely required for the induction of specific antibody responses to both TT and KLH in an antigen-induced in vitro assay. Monocytes were also required for the PWM induction of specific antibody in immunized subjects. Pulsing monocytes with specific antigen or with PWM consistently stimulated proliferation of T cells in absence of added antigen and could also stimulate specific antibody synthesis although less consistently. Stimulation of specific antibody responses with antigen required fewer numbers of monocytes than did stimulation of specific antibody responses with PWM. Polyclonal antibody synthesis induced by PWM was also dependent on monocytes. However, polyclonal antibody synthesis induced by supraoptimal concentrations of antigen was usually optimal in the absence of monocytes and was actually suppressed when increased numbers of monocytes were added to monocyte-depleted cultures. Monocyte supernatants could not replace the absolute requirements for monocytes in the induction of specific antibody synthesis. However, monocyte supernatants could profoundly modulate the antigen-specific as well as the polyclonal Ig response of lymphocytes to either antigen or PWM stimulation in a manner closely resembling monocytes themselves. Thus, we demonstrated that monocytes and their products play a critical role in the activation and immunoregulation of antigen-specific antibody responses of human B cells.


Emerging burdens of adolescent psychosocial health problems: a population-based study of 202 040 adolescents from 68 countries.
Psychosocial health problems are major public health burdens for adolescents. Identifying risk factors is essential to containing negative health implications. This study aimed to estimate the burden of psychosocial health problems among adolescents, and identify potential risk and protective factors. We used the Global School-based Student Health Survey data from 68 countries across six World Health Organization regions. We computed the overall, national- and regional-level weighted estimates of the mean number of psychosocial health problems. Adjusted Poisson regression models examined the factors associated with psychosocial health problems. Our sample totalled 202 040 adolescents aged 11-17 years (mean age 14.6 [s.d. 1.18] years), composed of 95 589 (47.31%) boys and 104 191 (51.57%) girls (2260 (1.12%) missing answers). Samoa had the highest mean number of psychosocial health problems (mean 2.64 [s.d. 1.41]), and Niue had the lowest (mean 0.33 [s.d. 0.72]). In the pooled analysis, the following factors were associated with higher rates of psychosocial health problems in adolescents globally: ≥13 years of age, being female, experiencing food insecurity, experiencing physical violence, having been bullied, engagement in physical altercation, experiencing serious injury, missing school without parental permission, lack of parental support or monitoring, parents who were not understanding of their child's problems and high sedentary behaviour. Being female, food insecurity, bullying, physical attacks or serious injury were all significantly associated with higher rates of psychosocial health problems in each of the six regions separately. The prevalence of psychosocial health problems is high among adolescents, although there are country-level variations. Health promotion strategies should consider the identified factors to promote adolescents' health and well-being.


Mutations in the rho transcription termination factor that affect RNA tracking.
Model studies have identified 16 conserved positively charged amino acids that form a positive strip pointing toward the center hole of Rho. Fourteen residues were individually changed to either an alanine or a glycine and one to a glutamate. Residues Arg(269), Arg(272), Lys(283), Arg(296), Lys(298), and Arg(299) form a subdomain (locus) located N-terminal to (above) the ATP hydrolysis domain (P-loop) and mutations in these residues led to either inactive Rho or to proteins displaying decreased k(cat) for poly(C)-dependent ATP hydrolysis, increased K(m) for ribo(C)(10) activation, and decreased transcription termination efficiencies (57-77%) compared with wild-type Rho. Residues Arg(347), Lys(348), Lys(352), and Arg(353) form a subdomain (locus) C-terminal to (below) the ATP hydrolysis domain, and mutations in these residues also show a decreased k(cat) for poly(C)-dependent ATP hydrolysis, an increased K(m) for ribo(C)(10) activation, and a 50-70% decrease in transcription termination, compared with wild-type Rho. Residues Arg(212) and Lys(336) surround the ATP hydrolysis domain, and mutations in these residues also altered the kinetic properties of Rho. We conclude that the secondary RNA-tracking site consists of amino acids whose putative orientation faces the central hole in Rho and in part reside in two clusters of positively charged residues located above and below the ATP hydrolysis domain.


Plasmapheresis Treatment of Hypertriglyceridemia-Induced Acute Pancreatitis: A Case Report.
Hypertriglyceridemia is the third most common etiology for acute pancreatitis (AP), after alcohol and gallstones. Clinical evidence is relatively weak in its support of plasmapheresis for the treatment of hypertriglyceridemia-induced acute pancreatitis (HTG-AP). We report a case of severe HTG-AP in a young man who was successfully treated with plasmapheresis. The patient achieved full resolution of symptoms within 48 hours from presentation and was discharged two days later. To our knowledge, no other report in literatures shows such dramatic response to plasmapheresis.


The Effect of Swelling Ratio on the Coulter Underestimation of Hydrogel Microsphere Diameters.
It has been demonstrated that the diameters of porous particles are underestimated by Coulter measurements. This phenomenon has also been observed in hydrogel particles, but not characterized. Since the Coulter principle uses the displacement of electrolyte to determine particle size, electrolyte contained within the swelled hydrogel microparticles results in an underestimate of actual particle diameters. The increased use of hydrogel microspheres in biomedical applications has led to the increased application of the Coulter principle to evaluate the size distribution of microparticles. A relationship between the swelling ratio of the particles and their reported Coulter diameters will permit calculation of the actual diameters of these particles. Using polyethylene glycol diacrylate hydrogel microspheres, we determined a correction factor that relates the polymer swelling ratio and the reported Coulter diameters to their actual size.


Oviductal extracellular vesicles (oviductosomes, OVS) are conserved in humans: murine OVS play a pivotal role in sperm capacitation and fertility.
Are extracellular vesicles (EVs) in the murine oviduct (oviductosomes, OVS) conserved in humans and do they play a role in the fertility of Pmca4-/- females? OVS and their fertility-modulating proteins are conserved in humans, arise via the apocrine pathway, and mediate a compensatory upregulation of PMCA1 (plasma membrane Ca2+-ATPase 1) in Pmca4-/- female mice during proestrus/estrus, to account for their fertility. Recently murine OVS were identified and shown during proestrus/estrus to express elevated levels of PMCA4 which they can deliver to sperm. PMCA4 is the major Ca2+ efflux pump in murine sperm and Pmca4 deletion leads to loss of sperm motility and male infertility as there is no compensatory upregulation of the remaining Ca2+ pump, PMCA1. Of the four family members of PMCAs (PMCA1-4), PMCA1 and PMCA4 are ubiquitous, and to date there have been no reports of one isoform being upregulated to compensate for another in any organ/tissue. Since Pmca4-/- females are fertile, despite the abundant expression of PMCA4 in wild-type (WT) OVS, we propose that OVS serve a role of packaging and delivering to sperm elevated levels of PMCA1 in Pmca4-/- during proestrus/estrus to compensate for PMCA4's absence. Fallopian tubes from pre-menopausal women undergoing hysterectomy were used to study EVs in the luminal fluid. Oviducts from sexually mature WT mice were sectioned after perfusion fixation to detect EVs in situ. Oviducts were recovered from WT and Pmca4-/- after hormonally induced estrus and sectioned for PMCA1 immunofluorescence (IF) (detected with confocal microscopy) and hematoxylin and eosin staining. Reproductive tissues, luminal fluids and EVs were recovered after induced estrus and after natural cycling for western blot analysis of PMCA1 and qRT-PCR of Pmca1 to compare expression levels in WT and Pmca4-/-. OVS, uterosomes, and epididymal luminal fluid were included in the comparisons. WT and Pmca4-/- OVS were analyzed for the presence of known PMCA4 partners in sperm and their ability to interact with PMCA1, via co-immunoprecipitation. In vitro uptake of PMCA1 from OVS was analyzed in capacitated and uncapacitated sperm via quantitative western blot analysis, IF localization and flow cytometry. Caudal sperm were also assayed for uptake of tyrosine-phosphorylated proteins which were shown to be present in OVS. Finally, PMCA1 and PMCA4 in OVS and that delivered to sperm were assayed for enzymatic activity. Human fallopian tubes were flushed to recover luminal fluid which was processed for OVS via ultracentrifugation. Human OVS were negatively stained for transmission electron microscopy (TEM) and subjected to immunogold labeling, to detect PMCA4. Western analysis was used to detect HSC70 (an EV biomarker), PMCA1 and endothelial nitric oxide synthase (eNOS) which is a fertility-modulating protein delivered to human sperm by prostasomes. Oviducts of sexually mature female mice were sectioned after perfusion fixation for TEM tomography to obtain 3D information and to distinguish cross-sections of EVs from those of microvilli and cilia. Murine tissues, luminal fluids and EVs were assayed for PMCA1 (IF and western blot) or qRT-PCR. PMCA1 levels from western blots were quantified, using band densities and compared in WT and Pmca4-/- after induced estrus and in proestrus/estrus and metestrus/diestrus in cycling females. In vitro uptake of PMCA1 and tyrosine-phosphorylated proteins was quantified with flow cytometry and/or quantitative western blot. Ca2+-ATPase activity in OVS and sperm before and after PMCA1 and PMCA4 uptake was assayed, via the enzymatic hydrolysis rate of ATP. TEM revealed that human oviducts contain EVs (exosomal and microvesicular). These EVs contain PMCA4 (immunolabeling), eNOS and PMCA1 (western blot) in their cargo. TEM tomography showed the murine oviduct with EV-containing blebs which typify the apocrine pathway for EV biogenesis. Western blots revealed that during proestrus/estrus PMCA1 was significantly elevated in the oviductal luminal fluid (OLF) (P = 0.02) and in OVS (P = 0.03) of Pmca4-/-, compared to WT. Further, while PMCA1 levels did not fluctuate in OLF during the cycle in WT, they were significantly (P = 0.02) higher in proestrus/estrus than at metestrus/diestrus in Pmca4-/-. The elevated levels of PMCA1 in proestrus/estrus, which mimics PMCA4 in WT, is OLF/OVS-specific, and is not seen in oviductal tissues, uterosomes or epididymal luminal fluid of Pmca4-/-. However, qRT-PCR revealed significantly elevated levels of Pmca1 transcript in Pmca4-/- oviductal tissues, compared to WT. PMCA1 could be transferred from OVS to sperm and the levels were significantly higher for capacitated vs uncapacitated sperm, as assessed by flow cytometry (P = 0.001) after 3 h co-incubation, quantitative western blot (P < 0.05) and the frequency of immuno-labeled sperm (P < 0.001) after 30 min co-incubation. Tyrosine phosphorylated proteins were discovered in murine OVS and could be delivered to sperm after their co-incubation with OVS, as detected by western, immunofluorescence localization, and flow cytometry. PMCA1 and PMCA4 in OVS were shown to be enzymatically active and this activity increased in sperm after OVS interaction. None. Although oviductal tissues of WT and Pmca4-/- showed no significant difference in PMCA1 levels, Pmca4-/- levels of OVS/OLF during proestrus/estrus were significantly higher than in WT. We have attributed this enrichment or upregulation of PMCA1 in Pmca4-/- partly to selective packaging in OVS to compensate for the lack of PMCA4. However, in the absence of a difference between WT and Pmca4-/- in the PMCA1 levels in oviductal tissues as a whole, we cannot rule out significantly higher PMCA1 expression in the oviductal epithelium that gives rise to the OVS as significantly higher Pmca1 transcripts were detected in Pmca4-/-. Since OVS and fertility-modulating cargo components are conserved in humans, it suggests that murine OVS role in regulating the expression of proteins required for capacitation and fertility is also conserved. Secondly, OVS may explain some of the differences in in vivo and in vitro fertilization for mouse mutants, as seen in mice lacking the gene for FER which is the enzyme required for sperm protein tyrosine phosphorylation. Our observation that murine OVS carry and can modulate sperm protein tyrosine phosphorylation by delivering them to sperm provides an explanation for the in vivo fertility of Fer mutants, not seen in vitro. Finally, our findings have implications for infertility treatment and exosome therapeutics. The work was supported by National Institute of Health (RO3HD073523 and 5P20RR015588) grants to P.A.M.-D. There are no conflicts of interests.


Reflex epilepsies: experience in Sri Lanka.
Reflex epilepsy (RE) is characterised by seizures that are regularly elicited by some specific stimulus or event mediated by neural pathways. In a prospective study of 1287 epileptic patients seen at Peradeniya, 223 (17.3%) were found to have RE, eating being the commonest stimulus (191 patients, 85.7%). Photosensitive epilepsy (PSE) was relatively rare. Intermittent photic stimulation on 874 unselected epileptic patients produced a positive photoconvulsive response in 60 (6.9%). None had photosensitive seizures, but 3 had a higher frequency of seizures while watching television. Eating epilepsy (EE) had the highest prevalence at Peradeniya (148/1000 epileptic patients). This group was male predominant, and the onset of epilepsy in most cases was in the second decade. The majority experienced partial complex seizures. Repetitive and chronic stimulation of the amygdala during eating is suggested as the mechanism underlying EE. Twenty-one patients had seizures evoked by calculation, problem solving or spatial tasks. Juvenile myoclonic epilepsy was the commonest form of seizure disorder in them. Although PSE itself is rare, self-induced epilepsy (SIE) was common. There were 8 patients who self-induced seizures. The majority were photosensitive and they induced seizures by gazing at the sun and waving a hand in front of the eyes. In the management of REs, clobazam produced impressive results. As for possible seizure-inhibitory mechanisms, our studies on a "Sathi" mediator showed definite EEG changes during mediation. Can mediation increase the seizure-threshold and abort or prevent the propagation of the epileptic discharge? The answer, apart from its possible therapeutic applications, may provide insight into the mechanisms of seizure generation.


PTH1R Actions on Bone Using the cAMP/Protein Kinase A Pathway.
After the initial signaling action of parathyroid hormone (PTH) on bone was shown to be activation of adenylyl cyclase, its target was found to be cells of the osteoblast lineage, to the exclusion of osteoclasts and their precursors. This led to the view that the osteoblast lineage regulated osteoclast formation, a proposal that was established when the molecular mechanisms of osteoclast formation were discovered. This is in addition to the effect of PTH1Rv signaling throughout the osteoblast differentiation process to favour the formation of bone-forming osteoblasts. Initial signaling in the PTH target cells through cAMP and protein kinase A (PKA) activation is extremely rapid, and marked by an amplification process in which the later event, PKA activation, precedes cAMP accumulation in time and is achieved at lower concentrations. All of this is consistent with the existence of "spare receptors", as is the case with several other peptide hormones. PTH-related protein (PTHrP), that was discovered as a cancer product, shares structural similarity with PTH in the amino-terminal domain that allows the hormone, PTH, and the autocrine/paracrine agent, PTHrP, to share actions upon a common G protein coupled receptor, PTH1R, through which they activate adenylyl cyclase with equivalent potencies. Studies of ligand-receptor kinetics have revealed that the PTH/PTH1R ligand-receptor complex, after initial binding and adenylyl cyclase activation at the plasma membrane, is translocated to the endosome, where adenylyl cyclase activation persists for a further short period. This behavior of the PTH1R resembles that of a number of hormones and other agonists that undergo such endosomal translocation. It remains to be determined whether and to what extent the cellular effects through the PTH1R might be influenced when endosomal is added to plasma membrane activation.


Morphology control and photocatalysis enhancement by the one-pot synthesis of carbon nitride from preorganized hydrogen-bonded supramolecular precursors.
We present an efficient synthesis of a modified carbon nitride photocatalyst by using supramolecular complexes of cyanuric acid, melamine, and 2,4-diamino-6-phenyl-1,3,5-triazine as precursors. We combined a self-templating approach for morphology control with the modification of photophysical properties by altering the chemical structure of the material. The resulting carbon nitrides exhibit high surface areas, defined morphologies, and a strong enhancement of light absorption in the visible-light region. A detailed analysis shows that the ratio changes of the three raw monomers resulted in different carbon nitride morphologies, absorption, and emission properties, along with the incorporation of different numbers of phenyl groups in the resulting carbon nitride structures. The modified carbon nitrides exhibit superior activity in the photodegradation of rhodamine B, up to 16 times that of bulk carbon nitride. The pyrolysis of rationally chosen supramolecular hydrogen-bonded precursors constitutes a synthetic pathway for the simple one-pot preparation of efficient, metal-free carbon nitride photocatalysts.


Treatment of acne scars with hyaluronic acid: an improved approach.
Acne scarring is a prevalent and challenging cosmetic issue, which is often addressed by multiple modalities. A low-viscosity non-animal stabilized hyaluronic acid (NASHA) dermal filler, injected in microdoses into the mid-to-superficial dermis, may provide a useful new approach to improving the appearance of depressed acne scars Twelve consecutive patients with moderate to severe acne scarring, who had completed a series of fractional laser resurfacing, underwent microinjections of 20 mg/mL hyaluronic acid (HA) gel into discrete depressed acne scars on the face Immediate visual improvement was observed in all lesions. The procedure was well tolerated. Adverse events were limited to transient pinpoint bleeding at the injection site Microinjection of low viscosity HA offers a valuable technique for the treatment of discrete depressed acne scars


PD-L1-directed PlGF/VEGF blockade synergizes with chemotherapy by targeting CD141+ cancer-associated fibroblasts in pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year overall survival rate. Patients with PDAC display limited benefits after undergoing chemotherapy or immunotherapy modalities. Herein, we reveal that chemotherapy upregulates placental growth factor (PlGF), which directly activates cancer-associated fibroblasts (CAFs) to induce fibrosis-associated collagen deposition in PDAC. Patients with poor prognosis have high PIGF/VEGF expression and an increased number of PIGF/VEGF receptor-expressing CAFs, associated with enhanced collagen deposition. We also develop a multi-paratopic VEGF decoy receptor (Ate-Grab) by fusing the single-chain Fv of atezolizumab (anti-PD-L1) to VEGF-Grab to target PD-L1-expressing CAFs. Ate-Grab exerts anti-tumor and anti-fibrotic effects in PDAC models via the PD-L1-directed PlGF/VEGF blockade. Furthermore, Ate-Grab synergizes with gemcitabine by relieving desmoplasia. Single-cell RNA sequencing identifies that a CD141+ CAF population is reduced upon Ate-Grab and gemcitabine combination treatment. Overall, our results elucidate the mechanism underlying chemotherapy-induced fibrosis in PDAC and highlight a combinatorial therapeutic strategy for desmoplastic cancers.


Role of peroxynitrite in secondary oxidative damage after spinal cord injury.
Peroxynitrite (PON, ONOO(-)), formed by nitric oxide synthase-generated nitric oxide radical ( NO) and superoxide radical (O(2) (-)), is a crucial player in post-traumatic oxidative damage. In the present study, we determined the spatial and temporal characteristics of PON-derived oxidative damage after a moderate contusion injury in rats. Our results showed that 3-nitrotyrosine (3-NT), a specific marker for PON, rapidly accumulated at early time points (1 and 3 h) and a significant increase compared with sham rats was sustained to 1 week after injury. Additionally, there was a coincident and maintained increase in the levels of protein oxidation-related protein carbonyl and lipid peroxidation-derived 4-hydroxynonenal (4-HNE). The peak increases of 3-NT and 4-HNE were observed at 24 h post-injury. In our immunohistochemical results, the co-localization of 3-NT and 4-HNE results indicates that PON is involved in lipid peroxidative as well as protein nitrative damage. One of the consequences of oxidative damage is an exacerbation of intracellular calcium overload, which activates the cysteine protease calpain leading to the degradation of several cellular targets including cytoskeletal protein (alpha-spectrin). Western blot analysis of alpha-spectrin breakdown products showed that the 145-kDa fragments of alpha-spectrin, which are specifically generated by calpain, were significantly increased as soon as 1 h following injury although the peak increase did not occur until 72 h post-injury. The later activation of calpain is most likely linked to PON-mediated secondary oxidative impairment of calcium homeostasis. Scavengers of PON, or its derived free radical species, may provide an improved antioxidant neuroprotective approach for the treatment of post-traumatic oxidative damage in the injured spinal cord.


Nucleotide sequence of the putative recognition site for coat protein in the RNAs of alfalfa mosaic virus and tobacco streak virus.
The sequence of the 3'-terminal 180 and 140 nucleotides of RNAs 2 and 3, respectively, of tobacco streak virus (TSV) was deduced by reverse transcription in the presence of a specific primer and chain terminators. Homology between the two RNAs was found to be restricted to a 3-terminal region of about 45 nucleotides. The data were compared with the sequence of the homologous region of 145 nucleotides occurring at the 3'-termini of the alfalfa mosaic virus (A1MV) RNAs, which contains the specific binding site for coat protein (Koper-Zwarthoff et al., Nucleic Acids Res. 7, 1887-1900 (1979); Houwing and Jaspars, Biochemistry 17, 2927-2933 (1978)). This was done because of the evidence that the RNAs of A1MV and TSV contain specific binding sites for their own as well as each others coat protein, and that binding of coat protein to these sites is required to initiate infection (Van Vloten-Doting, Virology 65, 215-225 (1975)). The 3'-terminal homologous regions of A1MV and TSV have two features in common: the presence of several stable hairpins and the multiple occurrence of the tetranucleotide sequence AUGC. The hairpins cause the linear array of tandemly repeated AUGC-boxes. It is postulated that the primary interaction of coat protein molecules with the RNAs of AlMV and TSV is a cooperative process involving several binding sites each being composed of a hairpin flanked at its 3'-side by an AUGC-sequence.


Bacterial community dynamics in a swine wastewater anaerobic reactor revealed by 16S rDNA sequence analysis.
Anaerobic digestion is a microbiological process of converting organic wastes into digestate and biogas in the absence of oxygen. In practice, disturbance to the system (e.g., organic shock loading) may cause imbalance of the microbial community and lead to digester failure. To examine the bacterial community dynamics after a disturbance, this study simulated an organic shock loading that doubled the chemical oxygen demand (COD) loading using a 4.5L swine wastewater anaerobic completely stirred tank reactor (CSTR). Before the shock (loading rate=0.65gCOD/L/day), biogas production rate was about 1-2L/L/day. After the shock, three periods representing increased biogas production rates were observed during days 1-7 (∼4.0L/L/day), 13 (3.3L/L/day), and 21-23 (∼6.1L/L/day). For culture-independent assessments of the bacterial community composition, the 454 pyrosequencing results indicated that the community contained >2500 operational taxonomic units (OTUs) and was dominated by three phyla: Bacteroidetes, Firmicutes, and Proteobacteria. The shock induced dynamic changes in the community composition, which was re-stabilized after approximately threefold hydraulic retention time (HRT). Intriguingly, upon restabilization, the community composition became similar to that observed before the shock, rather than reaching a new equilibrium.


Aggregative properties of different cell types of the fresh-water spongeEphydatia fluviatilis isolated on ficoll gradients.
Cell suspensions of the fresh-watersponge Ephydatia fluviatilis have been fractionated by means ofFicoll gradient centrifugation. Three fractions were isolated. The densest contains archeocyte-like cells only; the intermediate fraction is very rich in choanocytes, and the lightest is a mixture of cell types. Earch fraction shows specificaggregative properties and potentialities to reconstitute functional sponges.It appears that the sequence of reconstitution events can be selectively altered by certain disequilibria in the cell populationThese preliminary results constitute a first approach to the analysis ofcell type specificity in sponges.


The Fear of Positive Evaluation Scale: assessing a proposed cognitive component of social anxiety.
Cognitive-behavioral models propose that fear of negative evaluation is the core feature of social anxiety disorder. However, it may be that fear of evaluation in general is important in social anxiety, including fears of positive as well as negative evaluation. To test this hypothesis, we developed the Fear of Positive Evaluation Scale (FPES) and conducted analyses to examine the psychometric properties of the FPES, as well as test hypotheses regarding the construct of fear of positive evaluation (FPE). Responses from a large (n = 1711) undergraduate sample were utilized. The reliability, construct validity, and factorial validity of the FPES were examined; the distinction of FPE from fear of negative evaluation was evaluated utilizing confirmatory factor analysis; and the ability of FPE to predict social interaction anxiety above and beyond fear of negative evaluation was assessed. Results provide preliminary support for the psychometric properties of the FPES and the validity of the construct of FPE. The implications of FPE with respect to the study and treatment of social anxiety disorder are discussed.


Which nicotine products are gateways to regular use? First-tried tobacco and current use in college students.
The potential for emerging tobacco products (ETPs) to be gateway products for further tobacco use among youth is of significant concern. To examine use of various nicotine-containing products on a tobacco-free college campus and whether the first product tried predicts subsequent tobacco use. Undergraduate students (N=1,304) at a large university completed an online survey of past/current use of cigarettes; smokeless tobacco (SLT); hookah; ETPs (dissolvables, snus, and electronic cigarettes); and nicotine replacement therapy (NRT). Data were collected from September 2012 to May 2013 and analyses were conducted from June to September 2013. Students were classified as single, dual, or poly tobacco users. The sample consisted of 79.5% non-users, 13.8% single, 4.4% dual, and 1.5% poly users. Overall, 49.4% of participants reported trying a tobacco product. Hookah was the most tried product (38%), but cigarettes were most often the first product ever tried (51%). First product tried did not predict current tobacco use and non-use, but individuals who first tried SLT or cigarettes (rather than hookah or ETPs) were more likely to be poly tobacco users. Current tobacco users who first tried ETPs or hookah were largely non-daily users of hookah; current tobacco users who first tried cigarettes or SLT were largely non-daily or daily users of cigarettes/SLT. Hookah and ETPs are increasingly becoming the first tobacco product ever tried by youth; however, uptake of ETPs is poor, unlike cigarettes and SLT, and does not appear to lead to significant daily/non-daily use of cigarettes and SLT.


An overview of research advances in road traffic trauma in China.
With rapid development of social economies, road traffic accidents (RTAs) have continued to increase, and have become the "primary public hazard" to humans. Road traffic trauma (RTT) is a major cause of death in young people of all motorized countries. This article reviews the current advances in RTT research, in order to find some approaches to improving traffic administration and reducing RTAs and RTT. All available data were collected from government, literature, our own research, and conference proceedings. Statistical analysis from every country showed that human factors were still the main cause of RTAs, accounting for more than 90%. Vehicle and road factors caused 3-5% and less than 2% of the total RTAs, respectively. Approximately 85% of RTAs were caused by 21 to 45-year-olds. About 50% of deaths due to RTAs in the United States were related to drinking. In China, RTAs due to driver drinking accounted for 0.29-1.48%. About 6-8% of drivers were prone to RTA, causing 30-40% of RTAs. Seat belts are an effective way to prevent casualties, reducing mortality and morbidity by 13-50%. In China, about 70% of RTAs were related to bicycles. Prehospital emergency treatment is very important. About 35% of deaths may be avoided if the injured receive early and effective treatment. From 1983 to 1992 the mortality of RTT increased by 13% in 18 developing countries, while it decreased by 18% in 13 developed countries, indicating the importance of comprehensive treatment of traffic administration. In addition many advances have been made in basic scientific research of RTT, such as development of serial bioimpact machines and investigation of biomechanical and biochemical mechanisms of impact injuries. In this century, RTAs and RTT are predicted to continue to increase in many countries, especially in developing ones. Full cooperation and comprehensive treatment should be performed in order to improve traffic safety.


An investigation into four different sit-stand workstation use schedules.
Twelve office workers participated in a study investigating effects of four sit/stand schedules (90-min sit/30-min stand, 80/40, 105/15, and 60/60) via several objective and subjective measures (muscle fatigue, foot swelling, spinal shrinkage, and self-reported discomfort). Results showed that there were no significant differences in shoulder and low back static muscle activities between sitting and standing. Muscle fatigue was developed during workday under all schedules. The longest standing schedule seemed to have a tendency of reducing muscle fatigue. None of the schedules helped or worsened foot swelling and spinal shrinkage. More active break-time activities seemed reducing muscle fatigue and foot swelling. While the self-reported bodily discomfort levels were generally low, the preferred schedules among the participants were varied, although the least standing schedule was the least preferred. We may conclude that effects of using sit-stand workstation to improve musculoskeletal health may be limited but promoting more active break-time activities can help. Practitioner Summary: Sit-stand workstations are used to reduce work-related musculoskeletal disorders. This study shows that office workers prefer sit/stand durations in the range between 1:1 and 3:1. Longer standing may have the potential to reduce muscle fatigue. However, active break-time activities may be more effective in reducing muscle fatigue and foot swelling.


A synthetic resilin is largely unstructured.
Proresilin is the precursor protein for resilin, an extremely elastic, hydrated, cross-linked insoluble protein found in insects. We investigated the secondary-structure distribution in solution of a synthetic proresilin (AN16), based on 16 units of the consensus proresilin repeat from Anopheles gambiae. Raman spectroscopy was used to verify that the secondary-structure distributions in cross-linked AN16 resilin and in AN16 proresilin are similar, and hence that solution techniques (such as NMR and circular dichroism) may be used to gain information about the structure of the cross-linked solid. The synthetic proresilin AN16 is an intrinsically unstructured protein, displaying under native conditions many of the characteristics normally observed in denatured proteins. There are no apparent alpha-helical or beta-sheet features in the NMR spectra, and the majority of backbone protons and carbons exhibit chemical shifts characteristic of random-coil configurations. Relatively few peaks are observed in the nuclear Overhauser effect spectra, indicating that overall the protein is dynamic and unstructured. The radius of gyration of AN16 corresponds to the value expected for a denatured protein of similar chain length. This high degree of disorder is also consistent with observed circular dichroism and Raman spectra. The temperature dependences of the NH proton chemical shifts were also measured. Most values were indicative of protons exposed to water, although smaller dependences were observed for glycine and alanine within the Tyr-Gly-Ala-Pro sequence conserved in all resilins found to date, which is the site of dityrosine cross-link formation. This result implies that these residues are involved in hydrogen bonds, possibly to enable efficient self-association and subsequent cross-linking. The beta-spiral model for elastic proteins, where the protein is itself shaped like a spring, is not supported by the results for AN16. Both the random-network elastomer model and the sliding beta-turn model are consistent with the data. The results indicate a flat energy landscape for AN16, with very little energy required to switch between conformations. This ease of switching is likely to lead to the extremely low energy loss on deformation of resilin.


Surface properties of Indonesian-made narrow dynamic compression plates.
The enormous need of orthopaedic (surgical) implants such as osteosynthesis plates is difficult to be fulfilled in developing countries commonly rely on imported ones. One of the alternatives is utilization of local resources, but only after they have been proven safe to use, to overcome this problem. Surface properties are some of the determining factors of safety for those implants. We have succeeded in developing prototype of osteosynthesis plate and the results indicate that Indonesian-made plates need improvement with regards to the surface quality of physical characterization.


Demonstration of x-ray fluorescence imaging of a high-energy-density plasma.
Experiments at the Trident Laser Facility have successfully demonstrated the use of x-ray fluorescence imaging (XRFI) to diagnose shocked carbonized resorcinol formaldehyde (CRF) foams doped with Ti. One laser beam created a shock wave in the doped foam. A second laser beam produced a flux of vanadium He-α x-rays, which in turn induced Ti K-shell fluorescence within the foam. Spectrally resolved 1D imaging of the x-ray fluorescence provided shock location and compression measurements. Additionally, experiments using a collimator demonstrated that one can probe specific regions within a target. These results show that XRFI is a capable alternative to path-integrated measurements for diagnosing hydrodynamic experiments at high energy density.


Survey of 5 mm small-field output factor measurements in Australia.
The Australian Radiation Protection and Nuclear Safety Agency (ARPANSA) held a comparison exercise in April 2016 where participants came to ARPANSA and measured the output factor of a nominal 5 mm cone attached to the ARPANSA Elekta Synergy (Elekta, Crawley, UK) linear accelerator. The goal of the exercise was to compare the consistency and methods used by independent medical physicists in measuring small-field output factors. ARPANSA provided a three-dimensional scanning tank for detector setup and positioning, but the participants were required to measure the output factor with their own detectors. No information regarding output factors previously measured was supplied to participants to make each result as independent as possible. Fifteen groups travelled to ARPANSA bringing a wide range of detectors and methods. A total of 30 measurements of the output factor were made. The standard deviation of the measurements (excluding one expected outlier from an uncorrected ionization chamber measurement) was 3.6%. The results provide an insight into the consistency of small-field dosimetry being performed in Australia and New Zealand at the present time.


Helicobacter pylori-Negative Gastric Mucosa-Associated Lymphoid Tissue Lymphoma Presenting as Massive Gastrointestinal Bleed.
This case reports a patient that represents the minority of patients with gastric mucosa-associated lymphoid tissue (MALT) lymphoma who do not have underlying Helicobacter pylori gastritis. Gastric MALT lymphoma is a type of primary gastric lymphoma (PGL), which are extremely rare gastric malignancies characterized by proliferation of B-cells and infiltration of lymphoid tissue leading to destruction of gastric glands. Development of gastric MALT lymphoma is associated with H. pylori gastritis. Patients typically present with a wide range of symptoms including but not limited to epigastric pain, weight loss, gastrointestinal bleeding and gastric wall perforation. Gastric MALT lymphoma presenting as a massive gastrointestinal bleed is quite rare and only a few cases have been documented. Our case demonstrates that it is important to recognize that acute presentations of this disease may also occur.


Sensitivity and specificity of OraScan (R) toluidine blue mouthrinse in the detection of oral cancer and precancer.
The efficacy of 1% toluidine blue in the identification of oral malignancies and potentially malignant oral lesions was evaluated among a group of Asian patients (n = 102) with undiagnosed oral mucosal lesions and conditions (n = 145). The trial, utilising a ready-to-use kit, was controlled by histopathologic evaluation of a total of 87 dye-retained or dye-negative lesions. Eighteen oral carcinomas all retained the dye and there were no false negatives, yielding a test sensitivity of 100%. Eight of 39 oral epithelial dysplasias were toluidine blue-negative, giving a false negative rate of 20.5% and a sensitivity of 79.5% for oral epithelial dysplasias. The specificity of the technique was low (62%). Five dysplastic lesions were detected solely by the kit and this suggests that the method is valuable for surveillance of high-risk subjects in addition to its remarkable sensitivity in the detection of invasive carcinoma.


Engaging end-users in intervention research study design.
Coordinated Healthcare Interventions for Childhood Asthma Gaps in Outcomes (CHICAGO) Plan is a 3-arm multicenter pragmatic trial to evaluate asthma interventions in high-risk Chicago children presenting to emergency departments (ED) with asthma. A formative evaluation with end-users to provide input into the trial design and outcome instruments was conducted prior to trial initiation. A multi-level data gathering framework from the field of design and standard qualitative methods was employed. This included one focus group with asthma Community Health Workers (N = 8), two focus groups with caregivers of children with asthma (N = 9), in-home interviews with caregivers (N = 9), key informant interviews at six EDs and outpatient clinical sites (N = 19), and ED tours and observations (N = 6). Data were presented, discussed, and organized into themes. Data indicated that changes to the study design and discharge tool were warranted. A key insight was that ED discharge protocols typically place patient education at a single inopportune time, as families are preparing to leave the ED. At this point in time, families are less receptive to education due to fatigue and a desire to expedite the discharge process. The trial design was modified to reposition the discharge asthma plan to occur at earlier "teachable moments." Delivery of the asthma discharge plan was assigned to study-employed ED coordinators instead of ED providers and staff. Other potential challenges to study recruitment and implementation were raised and addressed. Engagement of end-users in the design phase of implementation research is critical to improve research feasibility and relevance.


Structural characterization of Au nano bipyramids: reshaping under thermal annealing, the capping agent effect and surface decoration with Pt.
Anisotropic gold nanoparticles offer potential applications due to their functionalities and shape-dependent properties. Reshaping noble metal nanoparticles is an interesting field with optical, surface-enhanced Raman spectroscopy, catalytic applications and potential application as a photothermic therapy. This work comprises a structural study on gold nano bipyramids (Au NBPs) and nanodumbbells, and the evolution of Au NBPs capped with cetyltrimethylammonium bromide and dodecanethiol through an in situ and ex situ heating process in high vacuum. Also, we study the reshaping of Au NBPs by the addition of Pt to study the surface modification and the strain generated on a single particle by geometric phase analysis.


Optimization of Dose Schedules for Chemotherapy of Early Colon Cancer Determined by High-Performance Computer Simulations.
Cancer chemotherapy dose schedules are conventionally applied intermittently, with dose duration of the order of hours, intervals between doses of days or weeks, and cycles repeated for weeks. The large number of possible combinations of values of duration, interval, and lethality has been an impediment to empirically determine the optimal set of treatment conditions. The purpose of this project was to determine the set of parameters for duration, interval, and lethality that would be most effective for treating early colon cancer. An agent-based computer model that simulated cell proliferation kinetics in normal human colon crypts was calibrated with measurements of human biopsy specimens. Mutant cells were simulated as proliferating and forming an adenoma, or dying if treated with cytotoxic chemotherapy. Using a high-performance computer, a total of 28 800 different parameter sets of duration, interval, and lethality were simulated. The effect of each parameter set on the stability of colon crypts, the time to cure a crypt of mutant cells, and the accumulated dose was determined. Of the 28 800 parameter sets, 434 parameter sets were effective in curing the crypts of mutant cells before they could form an adenoma and allowed the crypt normal cell dynamics to recover to pretreatment levels. A group of 14 similar parameter sets produced a minimal time to cure mutant cells. A different group of nine similar parameter sets produced the least accumulated dose. These parameter sets may be considered as candidate dose schedules to guide clinical trials for early colon cancer.


Sealing ability and adaptation of root-end filling materials in cavities prepared with different techniques.
The aim of this study was to evaluate the sealing ability and marginal adaptation of calcium silicate-based cements (CSCs) in root-end cavities prepared by ultrasonic and laser tips. A total of 72 extracted human maxillary incisor teeth were randomly divided as 60 teeth in experimental groups and 6 teeth each for positive and negative control groups. Specimens in experimental groups were obturated, their root-end resections were performed and randomly divided into six groups (n = 10) as follows: G1: Ultrasonic retrotip + MTA, G2: Ultrasonic retrotip + Calcium Enriched Mixture (CEM), G3: Ultrasonic retrotip + Biodentine, G4: Er:YAG laser tip + MTA, G5: Er:YAG laser tip + CEM, G6: Er:YAG laser tip + Biodentine. The sealing ability was measured by fluid transport method. Six specimens from each experimental group were randomly selected to analyze marginal adaptation and prepared for scanning electron microscopy (SEM) analysis. Micrographs were scored and also analyzed using Image J software. Data were analyzed with; two-way ANOVA, Bonferroni, Kruskall-Wallis, Mann-Whitney-U, Siegel & Castellan, and Spearman correlation coefficient tests. No significant difference was found between materials regarding the sealing ability and marginal adaptation (p > 0.05). Significantly greater fluid movement and poor marginal adaptation were seen for materials placed in cavities prepared by laser tips (p < 0.05). Positive correlation was found between the results of scoring and Image J analysis of SEM images (r = 0.596, p < 0.001). Fluid transport method and SEM analysis gave similar results suggesting the use of ultrasonic-retrotips for preparing root-end cavities which are going to be filled with one of these CSCs.


Beyond Structure: Defining the Function of the Gut Using Omic Approaches for Rational Design of Personalized Therapeutics.
Over the past 10 years, microbiome research has focused on defining the structures associated with different disease states in multiple systems, but has fallen short on showing causation. Prior omic studies have generated many new hypotheses, but moving forward we need to start dissecting the function of each bacterium alone and in concert with complex bacterial communities in well-characterized systems. Over the next 5 years, we need a merging of new omic technologies for exploratory studies with classical bacterial genetics, bacterial physiology, protein engineering, and biochemistry to further define the biochemical mechanisms of the gut microbiota. The future of the systems microbiology field will focus on targeted engineering and editing of the microbiome to alter function, which will be leveraged to prevent and/or treat human diseases. This perspective will focus on my contribution to the microbiome field, both past and present, and where I think research in the field is headed in the near future.


Maturation of human dendritic cells is accompanied by functional remodelling of the ubiquitin-proteasome system.
Dendritic cell maturation is the process by which immature dendritic cells differentiate into fully competent antigen-presenting cells that initiate T cell responses. Although some mechanistic aspects of DC maturation have begun to be characterised, very little is known about the genetic events regulating the ubiquitin-proteasome system which plays a key role at various levels of the immune response. Therefore, we here investigated the expression of more than 1000 genes related to the ubiquitin-proteasome system in maturing dendritic cells following various stimuli and identified a specific set of transcripts induced by lipopolysaccharide and/or Poly(I:C) which is largely distinct from that induced by CD40 ligand or pro-inflammatory cytokines. This group of genes was dependent on a type I interferon autocrine loop and included E1 and E2 enzymes, E3-ligases, de-ubiquitylating enzymes, proteasome components as well as the ubiquitin-like modifiers ISG15 and FAT10. We further demonstrate that the increased expression of the E2 enzyme UBE2L6 (UbcH8) is required for efficient antigen cross-presentation by dendritic cells. In summary, our data underline the importance of remodelling the ubiquitin-proteasome system for dendritic cell function.


Red cell ferritin and iron stores in chronic granulocytic leukemia.
Basic red cell ferritin was investigated in 28 patients with different phases of chronic granulocytic leukemia (CGL). Red cell ferritin was significantly decreased in remission after busulphan treatment and significantly elevated in the blast crisis as compared to healthy controls. Bone marrow stainable iron was decreased or absent in 86% of patients in the initial phase at the time of diagnosis and in 92% of those in remission. Red cell ferritin correlated with serum ferritin, however, serum ferritin level remained above normal range during all phases of the disease. A negative correlation between red cell ferritin and hemoglobin (Hb) (r = -0.605, p < 0.001) suggested that red cell ferritin level reflected the rate of iron utilization for heme synthesis. Decreased red cell iron stores observed in the remission may be explained by regression of dyserythropoiesis and by restoration of normal Hb synthesis after busulphan treatment. A progressive dyserythropoiesis in the blast crisis may lead to an increased red cell ferritin level.


A Comprehensive Review of Liver Allograft Fibrosis and Steatosis: From Cause to Diagnosis.
Despite advances in posttransplant care, long-term outcomes for liver transplant recipients remain unchanged. Approximately 25% of recipients will advance to graft cirrhosis and require retransplantation. Graft fibrosis progresses in the context of de novo or recurrent disease. Recurrent hepatitis C virus infection was previously the most important cause of graft failure but is now curable in the majority of patients. However, with an increasing prevalence of obesity and diabetes and nonalcoholic fatty liver disease as the most rapidly increasing indication for liver transplantation, metabolic dysfunction-associated liver injury is anticipated to become an important cause of graft fibrosis alongside alloimmune hepatitis and alcoholic liver disease. To better understand the landscape of the graft fibrosis literature, we summarize the associated epidemiology, cause, potential mechanisms, diagnosis, and complications. We additionally highlight the need for better noninvasive methods to ameliorate the management of graft fibrosis. Some examples include leveraging the microbiome, genetic, and machine learning methods to address these limitations. Overall, graft fibrosis is routinely seen by transplant clinicians, but it requires a better understanding of its underlying biology and contributors that can help inform diagnostic and therapeutic practices.


Shift of graft-versus-host-disease target organ tropism by dietary vitamin A.
Gut-homing of donor T cells is causative for the development of intestinal GvHD in recipients of allogeneic hematopoietic stem cell transplantation (HSCT). Expression of the gut-specific homing receptors integrin-α4β7 and chemokine receptor CCR9 on T cells is imprinted in gut-associated lymphoid tissues (GALT) under the influence of the vitamin A metabolite retinoic acid. Here we addressed the role of vitamin A deficiency in HSCT-recipients for donor T cell migration in the course of experimental GvHD. Vitamin A-deficient (VAD) mice were prepared by feeding them a vitamin A-depleted diet. Experiments were performed in a C57BL/6 into BALB/c model of acute GvHD. We found that expression of integrin-α4β7 and CCR9 in GALT was reduced in VAD recipients after HSCT. Competitive in vivo homing assays showed that allogeneic T cells primed in VAD mice did not home as efficiently to the intestine as T cells primed in mice fed with standard diet (STD). The course of GvHD was ameliorated in VAD HSCT-recipients and, consequently, their survival was prolonged compared to recipients receiving STD. However, VAD-recipients were not protected and died of clinical GvHD. We found reduced numbers of donor T cells in the intestine but increased cell counts and tissue damage in other organs of VAD-recipients. Furthermore, we observed high IFN-γ(+)CD4(+) and low FoxP3(+)CD4(+) frequencies of total donor CD4(+) T cells in VAD as compared to STD recipients. Taken together, these results indicate that dietary vitamin A deficiency in HSCT-recipients changed target organ tropism in GvHD but also resulted in fatal inflammation after HSCT.