Dynamics of an ant-ant obligate mutualism: colony growth, density dependence and frequency dependence.
In insect societies, worker vs. queen development (reproductive caste) is typically governed by environmental factors, but many Pogonomyrmex seed-harvester ants exhibit strict genetic caste determination, resulting in an obligate mutualism between two reproductively isolated lineages. Same-lineage matings produce fertile queens while alternate-lineage matings produce sterile workers. Because new virgin queens mate randomly with multiple males of each lineage type, and both worker and queen phenotypes are required for colony growth and future reproduction, fitness is influenced by the relative frequency of each lineage involved in the mutualistic breeding system. While models based solely on frequency-dependent selection predict the convergence of lineage frequencies towards equal (0.5/0.5), we surveyed the lineage ratios of 49 systems across the range of the mutualism and found that the global lineage frequency differed significantly from equal. Multiple regression analysis of our system survey data revealed that the density and relative frequency of one lineage decreases at lower elevations, while the frequency of the alternate lineage increases with total colony density. While the production of the first worker cohort is largely frequency dependent, relying on the random acquisition of worker-biased sperm stores, subsequent colony growth is independent of lineage frequency. We provide a simulation model showing that a net ecological advantage held by one lineage can lead to the maintenance of stable but asymmetric lineage frequencies. Collectively, these findings suggest that a combination of frequency-dependent and frequency-independent mechanisms can generate many different localized and independently evolving system equilibria.


Source impacts by volatile organic compounds in an industrial city of southern Taiwan.
This study investigates source impacts by airborne volatile organic compounds (VOC) at two sites designated for traffic and industry, in the largest industrial area Kaohsiung, southern Taiwan. The samples were collected at the two sites simultaneously during rush and non-rush hours in summer and autumn seasons. Same pattern of VOC groups were found at both sites: most abundant aromatics (78-95%) followed by alkanes (2-16%) and alkenes (0-6%). The BTEX concentration measured at the two sites ranged from 69 to 301 ppbC. Toluene, isopentane, ethylbenzene, and benzene were found to be the most abundant species. Speciation of VOCs was characterized with several skills including principal component factor analysis and BTEX characteristic ratios. Each of the resulted principal factors at the two sites explained over 80% of the VOCs data variance, and indicated that both of the sampling sites were influenced by both traffic and industrial sources with separately different levels. The remarkable patterns of the first two factors described not only the similarity but also the discrepancy at the two sampling sites, in terms of the source impacts. The high T/B ratios (7.56-14.25) observed at the industrial site implied the important impact from mobile emissions. The indicators, m,p-xylene/benzene and o-xylene/benzene, also confirmed the potential source of motor vehicles at both of the sampling sites. Air age assessment showed that more than half of the total observations located in the domain of fresh air. Low X/E ratios implied somewhat aged air mass transported to the sampling sites. The industrial site might not only encounter emissions from the industry sources, but also under unavoidable impact from the traffic sources.


Cadherin-6 mediates the heterotypic interactions between the hemopoietic osteoclast cell lineage and stromal cells in a murine model of osteoclast differentiation.
Osteoclasts are multinucleated cells of hemopoietic origin that are responsible for bone resorption during physiological bone remodeling and in a variety of bone diseases. Osteoclast development requires direct heterotypic cell-cell interactions of the hemopoietic osteoclast precursors with the neighboring osteoblast/stromal cells. However, the molecular mechanisms underlying these heterotypic interactions are poorly understood. We isolated cadherin-6 isoform, denoted cadherin-6/2 from a cDNA library of human osteoclast-like cells. The isolated cadherin-6/2 is 3,423 bp in size consisting of an open reading frame of 2,115 bp, which encodes 705 amino acids. This isoform lacks 85 amino acids between positions 333 and 418 and contains 9 different amino acids in the extracellular domain compared with the previously described cadherin-6. The human osteoclast-like cells also expressed another isoform denoted cadherin-6/1 together with the cadherin-6. Introduction of cadherin-6/2 into L-cells that showed no cell-cell contact caused evident morphological changes accompanied with tight cell-cell association, indicating the cadherin-6/2 we isolated here is functional. Moreover, expression of dominant-negative or antisense cadherin-6/2 construct in bone marrow-derived mouse stromal ST2 cells, which express only cadherin-6/2, markedly impaired their ability to support osteoclast formation in a mouse coculture model of osteoclastogenesis. Our results suggest that cadherin-6 may be a contributory molecule to the heterotypic interactions between the hemopoietic osteoclast cell lineage and osteoblast/bone marrow stromal cells required for the osteoclast differentiation. Since both osteoclasts and osteoblasts/bone marrow stromal cells are the primary cells controlling physiological bone remodeling, expression of cadherin-6 isoforms in these two cell types of different origin suggests a critical role of these molecules in the relationship of osteoclast precursors and cells of osteoblastic lineage within the bone microenvironment.


Polyelectrolyte Influence on Beta-Hairpin Peptide Stability: A Simulation Study.
Assemblies of proteins and charged macromolecules (polyelectrolytes) find important applications as pharmaceutical formulations, biocatalysts, and cell-contacting substrates. A key question is how the polymer component influences the structure and function of the protein. The present paper addresses the influence of charged polymers on the thermal stability of two model beta-hairpin-forming peptides through an all-atom, replica exchange molecular dynamics simulation. The (negatively charged) peptides consist of the terminal 16 amino acids of the B1 domain of Protein G (GB1) and a variant with three of the GB1 residues substituted with tryptophan (Tryptophan Zipper 4, or TZ4). A (cationic) lysine polymer is seen to thermally stabilize TZ4 and destabilize GB1, while a (also cationic) chitosan polymer slightly stabilizes GB1 but has essentially no effect on TZ4. Free energy profiles reveal folded and unfolded conformations to be separated by kinetic barriers generally acting in the direction of the thermodynamically favored state. Through application of an Ising-like statistical mechanical model, a mechanism is proposed based on competition between (indirect) entropic stabilization of folded versus unfolded states and (direct) competition for hydrogen-bonding and hydrophobic interactions. These findings have important implications to the design of polyelectrolyte-based materials for biomedical and biotechnological applications.


Mapping temperature-sensitive mutations at a genome scale to engineer growth switches in Escherichia coli.
Temperature-sensitive (TS) mutants are a unique tool to perturb and engineer cellular systems. Here, we constructed a CRISPR library with 15,120 Escherichia coli mutants, each with a single amino acid change in one of 346 essential proteins. 1,269 of these mutants showed temperature-sensitive growth in a time-resolved competition assay. We reconstructed 94 TS mutants and measured their metabolism under growth arrest at 42°C using metabolomics. Metabolome changes were strong and mutant-specific, showing that metabolism of nongrowing E. coli is perturbation-dependent. For example, 24 TS mutants of metabolic enzymes overproduced the direct substrate metabolite due to a bottleneck in their associated pathway. A strain with TS homoserine kinase (ThrBF267D ) produced homoserine for 24 h, and production was tunable by temperature. Finally, we used a TS subunit of DNA polymerase III (DnaXL289Q ) to decouple growth from arginine overproduction in engineered E. coli. These results provide a strategy to identify TS mutants en masse and demonstrate their large potential to produce bacterial metabolites with nongrowing cells.


[Treatment of myelodysplastic syndrome by combined traditional Chinese medicine and Western medicine therapy].
50 cases were treated with Myelodysplastic Syndrome (MDS) by combined TCM-WM therapy. They were classified into RA 17 cases, RAS 6, RAEB 19, CMML 1 and RAEBT 7. The patients were divided into two groups, one with RA and RAS receiving treatment of hemopoietic and immune drugs plus Chinese medicinal herbs, the other with RAEB, CMML and RAEBT receiving treatment of LD Ara-c and LD Hom chemotherapy plus medicinal herbs. The effective rates were 47.83% and 62.96% respectively, the total effective rate being 56%. 6 cases (RAEB 4, RA 1, RAS 1) were treated with all-trans retinoic acid used as an inducer of differentiation, 2 of them were effective. 11 patients with MDS who had transformed into acute leukemia were treated by LD Ara-c and combined TCM-WM chemotherapy, the remission rate was 54.55% and the survival period was 9-27 months after remission. In some cases low dose chemotherapy resulted in hemocytopenia, bone marrow inhibition, infection, mild nausea and anorexia.


New Opportunities for Advancing Dyadic Health Science in Gerontology.
As dyadic health science enters a golden age, important conceptual, theoretical, and technical challenges remain. This forum brings together perspectives on the burgeoning dyadic literature from several subdisciplines within aging research. We first define key concepts and terms so that interested researchers can navigate the complex and various ways in which dyadic health research is conducted. We discuss exciting scientific advances and close by identifying crucial challenges and considerations that coincide with important future directions for the field.


Videoscope-Assisted Minimally Invasive Periodontal Surgery: One-Year Outcome and Patient Morbidity.
The aim of this study was to report the 1-year clinical outcomes from videoscope-assisted minimally invasive surgery (V-MIS). A sample of 18 patients having sites with residual pocket probing depth (PPD) of at least 5 mm and 2 mm loss in clinical attachment level (CAL) following initial nonsurgical therapy were treated with V-MIS. At 12 months postsurgery, there was a statistically significant improvement (P < .001) in mean PPD (4.11 ± 0.98 mm) and CAL (4.58 ± 1.19 mm) in all surgical sites. A mean improvement in soft tissue height (0.48 ± 0.65 mm, P = .006) was also observed. In most cases, patients reported no postoperative discomfort. The improvements associated with V-MIS appear to be favorable when compared with previously reported results of periodontal regenerative surgery. The lack of postsurgical recession following V-MIS has not been reported with traditional regenerative surgery.


Acute treatment with valproic acid and l-thyroxine ameliorates clinical signs of experimental autoimmune encephalomyelitis and prevents brain pathology in DA rats.
Multiple sclerosis (MS) is the most common chronic inflammatory demyelinating disease of the central nervous system (CNS) in young adults. Chronic treatments with histone deacetylase inhibitors (HDACis) have been reported to ameliorate experimental autoimmune encephalomyelitis (EAE), a rodent model of MS, by targeting immune responses. We have recently shown that the HDAC inhibition/knockdown in the presence of thyroid hormone (T3) can also promote oligodendrocyte (OL) differentiation and expression of myelin genes in neural stem cells (NSCs) and oligodendrocyte precursors (OPCs). In this study, we found that treatment with an HDACi, valproic acid (VPA), and T3, alone or in combination, directly affects encephalitogenic CD4+ T cells. VPA, but not T3, compromised their proliferation, while both molecules reduced the frequency of IL-17-producing cells. Transfer of T3, VPA and VPA/T3 treated encephalitogenic CD4+ T cells into naïve rats induced less severe EAE, indicating that the effects of these molecules are persistent and do not require their maintenance after the initial stimuli. Thus, we investigated the effect of acute treatment with VPA and l-thyroxine (T4), a precursor of T3, on myelin oligodendrocyte glycoprotein-induced EAE in Dark Agouti rats, a close mimic of MS. We found that a brief treatment after disease onset led to sustained amelioration of EAE and prevention of inflammatory demyelination in the CNS accompanied with a higher expression of myelin-related genes in the brain. Furthermore, the treatment modulated immune responses, reduced the number of CD4+ T cells and affected the Th1 differentiation program in the brain. Our data indicate that an acute treatment with VPA and T4 after the onset of EAE can produce persistent clinically relevant therapeutic effects by limiting the pathogenic immune reactions while promoting myelin gene expression.


Meticulous endoscopic sinus surgery to prevent recurrence of massive nasal polyposis.
To evaluate the effect of meticulous endoscopic surgery, including opening of all involved sinus cells, on the subsequent symptoms and endoscopic findings of patients with massive nasal polyposis. One hundred patients with massive nasal polyposis resistant to medical treatment were selected. We documented each patient's demographic data, associated diseases, endoscopic findings, Lund-Mackay score and Sino-Nasal Outcome Test 22 (SNOT22) symptom score. All patients were followed up for at least two years to evaluate any recurrence. Of the 100 patients, 20 per cent had a history of asthma and 27 per cent had undergone previous surgery. All underwent endoscopic sinus surgery. After two years of follow up, 8 per cent had recurrence requiring surgery. Recurrence was significantly associated with a history of asthma (p < 0.001) and the histopathological presence of eosinophilia (p = 0.014). Meticulous endoscopic opening of all involved sinus cells can be a safe and effective means of controlling massive nasal polyposis, with an acceptable recurrence rate.


Diagnosis of acute respiratory distress syndrome (DARTS) by bedside exhaled breath octane measurements in invasively ventilated patients: protocol of a multicentre observational cohort study.
Acute respiratory distress syndrome (ARDS) is currently diagnosed by the Berlin Definition. Diagnosis is subjective and often late. Untargeted metabolomics analysis of exhaled breath with gas chromatography and mass spectrometry (GC-MS) showed that the breath concentration of octane has a high diagnostic accuracy for ARDS. To facilitate rapid bedside measurement of this biomarker, a point-of-care (POC) breath test was developed. A prototype already showed good reproducibility and repeatability for the detection of octane. In this study we aim to measure octane in exhaled breath of invasively ventilated intensive care unit (ICU) patients and validate the diagnostic accuracy of the breath test for the early diagnosis of ARDS. This is a multicentre observational cohort study in patients admitted to the ICU receiving invasive ventilation for at least 24 hours. At least 500 patients in two academic hospitals in The Netherlands will be included. ARDS patients will be compared to patients without ARDS. ARDS diagnosis will be based on the Berlin Definition. Two diagnostic assessments will be performed during the first 72 hours of invasive ventilation, including breath sampling, arterial blood gas analysis and lung ultrasound (LUS). In patients fulfilling the criteria for ARDS, three additional breath samples will be taken to assess resolution. The primary endpoint is the diagnostic accuracy for ARDS, defined by the area under the receiver operating characteristics curve (AUROCC) of octane concentration in exhaled breath. Secondary endpoints are the association between exhaled breath octane and ARDS adjusted for confounders, and the added diagnostic accuracy of the breath test on top of the Lung Injury Prediction Score (LIPS). This is the first study that validates a metabolic biomarker of ARDS in an adequate sample size. The major novelty is the use of a POC breath test that has been specifically developed for the purpose of diagnosing ARDS. Strengths are; assessment in the early phase, in patients at risk for ARDS, longitudinal sampling and an expert panel to reliably diagnose ARDS. This study will provide a decisive answer on the question if exhaled breath metabolomics can be used to diagnose ARDS. The trial is registered at trialregister.nl (ID: NL8226) with the tag "DARTS".


Pattern and motion vision in cats with selective loss of cortical directional selectivity.
Neurons in the visual cortex of cats reared in 8 Hz stroboscopic illumination show a profound loss of directional selectivity, but no detectable deficits in orientation selectivity, contrast sensitivity, and temporal frequency response, and only a slight reduction in spatial resolution. In the present study, spatial vision, temporal resolution, and a variety of motion detection and discrimination thresholds were examined behaviorally in such cats. These psychophysical measurements revealed nearly normal spatial and temporal vision, but severe abnormalities in visual discriminations based on differences in stimulus direction. Specifically, strobe-reared cats showed normal orientation discrimination and temporal frequency resolution, nearly normal contrast sensitivity at low spatial frequencies, and a slight reduction of sensitivity to high spatial frequencies. At high contrasts, the cats were able to discriminate opposite directions of motion over a wide range of visible speeds, and their performance was indistinguishable from that of normal cats. However, a comparison of contrast thresholds for detecting moving gratings and for discriminating their direction of motion revealed severe abnormalities in strobe-reared animals. At low spatial frequencies (0.28 cycles/deg), normal cats could discriminate the direction of grating motion at contrasts that were just barely visible, whereas the strobe-reared cats could detect the grating at contrasts similar to those required by normal cats, but required contrasts about 10 X the threshold to identify the direction of motion. Normal cats showed nearly identical contrast sensitivity for detecting and discriminating gratings of high spatial frequency at high temporal frequency (drift rates), but when the temporal frequency was low, their sensitivity for detection exceeded that for direction discrimination.(ABSTRACT TRUNCATED AT 250 WORDS)


A 58-base-pair region of the human C3 gene confers synergistic inducibility by interleukin-1 and interleukin-6.
We have cloned the promoter for the human third component of complement (C3) gene and have identified sequences involved in its regulation during the acute-phase response. A construct linking 199 bp of the C3 promoter to the firefly luciferase gene was found to be very responsive to interleukin-1 (IL-1) and modestly responsive to interleukin-6 (IL-6) by transfection analysis in the human hepatoma line Hep3B2. Simultaneous treatment with the two cytokines showed a strong synergy between the actions of the two molecules. A 58-bp fragment (-127 to -70 bp) was shown by 5' and 3' deletional mutagenesis to contain cis-acting elements that mediated both the IL-1 response and the IL-1-plus-IL-6 synergistic response of this promoter. When coupled to a heterologous promoter, this fragment enabled the synergistic induction by IL-1 plus IL-6. Sequences homologous to the palindrome ACATTGCACAATCT, which mediates the induction of the IL-6 gene by IL-1 (S. Akira, H. Isshiki, T. Sugita, O. Tanabe, S. Kinoshita, Y. Nishio, T. Nakajima, T. Hirano, and T. Kishimoto, EMBO J. 9:1897-1906, 1990), and the core sequence of the IL-6-responsive element of the rat alpha 2-macroglobulin gene (CTGGGA; M. Hattori, L. J. Abraham, W. Northemann, and G. H. Fey, Proc. Natl. Acad. Sci. USA 87:2364-2368, 1990) are contained within this fragment in immediate juxtaposition and partially overlapping. Site-directed mutagenesis within this homology region drastically reduced the inducibility of the C3 promoter by either cytokine. DNase I footprinting analysis defined a binding site for the transcription factor CCAAT/enhancer-binding protein (C/EBP), which included the IL-1-responsive element-like sequence. No differences were seen between the footprints generated by using extracts from unstimulated and IL-1-stimulated Hep3B2 cells. However, gel retardation analyses revealed two IL-1-specific bands. The data suggest that the induction by IL-1 is mediated by a factor belonging to the family of C/EBP-related proteins.


Comparison of ligase chain reaction and culture for detection of Neisseria gonorrhoeae in genital and extragenital specimens.
In addition to the urogenital tract, Neisseria gonorrhoeae infects extragenital sites such as the pharynx and anorectal canal. Culture and a ligase chain reaction (LCR)-based assay were compared for their performance for the diagnosis of N. gonorrhoeae infection with specimens from various urogenital and extragenital sites of 200 men and 125 women. The sensitivity and specificity of the LCR assay with male urethral swabs were both 100%, compared to values of 95.9 and 100%, respectively, for culture of urethral swabs or 98.0 and 100%, respectively, for LCR with first-void urine (FVU). For women, LCR with FVU showed the highest sensitivity (94.7%), and culture of urethral samples showed the lowest sensitivity (63.2%) (P < 0.05). In a selected subgroup of 47 men and 22 women at increased risk, the rates of pharyngeal infection were 15 and 18%, respectively, and those of anorectal infection were 13 and 45%, respectively. The sensitivity of LCR was greater than that of culture for both pharyngeal and anorectal specimens. Thus, the overall performance of LCR testing with swabs or FVU was better than that of culture for the diagnosis of genital or extragenital gonorrhea.


Abnormal renal hemodynamics in black salt-sensitive patients with hypertension.
African-Americans with essential hypertension are more prone to the development of renal failure and are frequently salt-sensitive as well. Because alterations of intrarenal hemodynamics are important in the progression of renal disease and because salt-sensitive animal models with hypertension manifest a greater propensity to develop glomerulosclerosis in association with a rise in glomerular capillary pressure, we tested whether the renal hemodynamic adaptation to high dietary Na+ intake differs in salt-sensitive and salt-resistant hypertensive patients. We studied 17 black and nine white patients with essential hypertension who were placed on a low Na+ diet (20 meq/day) for 9 days, followed by a high Na+ diet (200 meq/day) for 14 days. During the last 4 days of each diet regimen, they received 30 mg/day of slow-release nifedipine. Eleven blacks were salt-sensitive, and all whites were salt-resistant. During the low Na+ diet period, salt-sensitive and salt-resistant patients had similar mean arterial pressure, glomerular filtration rate, effective renal plasma flow, and filtration fraction.(ABSTRACT TRUNCATED AT 250 WORDS)


A novel mutation in the lysyl hydroxylase 1 gene causes decreased lysyl hydroxylase activity in an Ehlers-Danlos VIA patient.
The clinical diagnosis of a patient with the phenotype of Ehlers-Danlos syndrome type VI was confirmed biochemically by the severely diminished level of lysyl hydroxylase (LH) activity in the patient's skin fibroblasts. A novel homozygous mutation, a single base change of T(1360)-->G in exon 13 of the LH1 gene, predicted to result in W446G, was identified in the patient's full-length cDNA. This was confirmed in genomic DNA from both the patient and her parents, who were heterozygous for the mutation. This mutation was introduced into an LH1-pAcGP67 baculoviral construct and expressed, in parallel with normal LH1, in an insect cell system. The loss of LH activity in the mutated recombinant construct confirmed the pathogenicity of this mutation. Although not in the major catalytic site, this mutation occurs in a highly conserved region of the LH1 gene and may contribute to loss of activity by interfering with normal folding of the enzyme.


Earthworms do not increase greenhouse gas emissions (CO2 and N2O) in an ecotron experiment simulating a three-crop rotation system.
Earthworms are known to stimulate soil greenhouse gas (GHG) emissions, but the majority of previous studies have used simplified model systems or lacked continuous high-frequency measurements. To address this, we conducted a 2-year study using large lysimeters (5 m2 area and 1.5 m soil depth) in an ecotron facility, continuously measuring ecosystem-level CO2, N2O, and H2O fluxes. We investigated the impact of endogeic and anecic earthworms on GHG emissions and ecosystem water use efficiency (WUE) in a simulated agricultural setting. Although we observed transient stimulations of carbon fluxes in the presence of earthworms, cumulative fluxes over the study indicated no significant increase in CO2 emissions. Endogeic earthworms reduced N2O emissions during the wheat culture (- 44.6%), but this effect was not sustained throughout the experiment. No consistent effects on ecosystem evapotranspiration or WUE were found. Our study suggests that earthworms do not significantly contribute to GHG emissions over a two-year period in experimental conditions that mimic an agricultural setting. These findings highlight the need for realistic experiments and continuous GHG measurements.


Evaluating Clinical Genome Sequence Analysis by Watson for Genomics.
Background: Oncologists increasingly rely on clinical genome sequencing to pursue effective, molecularly targeted therapies. This study assesses the validity and utility of the artificial intelligence Watson for Genomics (WfG) for analyzing clinical sequencing results. Methods: This study identified patients with solid tumors who participated in in-house genome sequencing projects at a single cancer specialty hospital between April 2013 and October 2016. Targeted genome sequencing results of these patients' tumors, previously analyzed by multidisciplinary specialists at the hospital, were reanalyzed by WfG. This study measures the concordance between the two evaluations. Results: In 198 patients, in-house genome sequencing detected 785 gene mutations, 40 amplifications, and 22 fusions after eliminating single nucleotide polymorphisms. Breast cancer (n = 40) was the most frequent diagnosis in this analysis, followed by gastric cancer (n = 31), and lung cancer (n = 30). Frequently detected single nucleotide variants were found in TP53 (n = 107), BRCA2 (n = 24), and NOTCH2 (n = 23). MYC (n = 10) was the most frequently detected gene amplification, followed by ERBB2 (n = 9) and CCND1 (n = 6). Concordant pathogenic classifications (i.e., pathogenic, benign, or variant of unknown significance) between in-house specialists and WfG included 705 mutations (89.8%; 95% CI, 87.5%-91.8%), 39 amplifications (97.5%; 95% CI, 86.8-99.9%), and 17 fusions (77.3%; 95% CI, 54.6-92.2%). After about 12 months, reanalysis using a more recent version of WfG demonstrated a better concordance rate of 94.5% (95% CI, 92.7-96.0%) for gene mutations. Across the 249 gene alterations determined to be pathogenic by both methods, including mutations, amplifications, and fusions, WfG covered 84.6% (88 of 104) of all targeted therapies that experts proposed and offered an additional 225 therapeutic options. Conclusions: WfG was able to scour large volumes of data from scientific studies and databases to analyze in-house clinical genome sequencing results and demonstrated the potential for application to clinical practice; however, we must train WfG in clinical trial settings.


Patient- and provider-reported information about transplantation and subsequent waitlisting.
Because informed consent requires discussion of alternative treatments, proper consent for dialysis should incorporate discussion about other renal replacement options including kidney transplantation (KT). Accordingly, dialysis providers are required to indicate KT provision of information (KTPI) on CMS Form-2728; however, provider-reported KTPI does not necessarily imply adequate provision of information. Furthermore, the effect of KTPI on pursuit of KT remains unclear. We compared provider-reported KTPI (Form-2728) with patient-reported KTPI (in-person survey of whether a nephrologist or dialysis staff had discussed KT) in a prospective ancillary study of 388 hemodialysis initiates. KTPI was reported by both patient and provider for 56.2% of participants, by provider only for 27.8%, by patient only for 8.3%, and by neither for 7.7%. Among participants with provider-reported KTPI, older age was associated with lack of patient-reported KTPI. Linkage with the Scientific Registry for Transplant Recipients showed that 20.9% of participants were subsequently listed for KT. Patient-reported KTPI was independently associated with a 2.95-fold (95% confidence interval [95% CI], 1.54 to 5.66; P=0.001) higher likelihood of KT listing, whereas provider-reported KTPI was not associated with listing (hazard ratio, 1.18; 95% CI, 0.60 to 2.32; P=0.62). Our findings suggest that patient perception of KTPI is more important for KT listing than provider-reported KTPI. Patient-reported and provider-reported KTPI should be collected for quality assessment in dialysis centers because factors associated with discordance between these metrics might inform interventions to improve this process.


Cardiorespiratory fitness and physical activity in Luo, Kamba, and Maasai of rural Kenya.
Although habitual physical activity energy expenditure (PAEE) and cardio-respiratory fitness (CRF) are now well-established determinants of metabolic disease, there is scarcity of such data from Africa. The aim of this study was to describe objectively measured PAEE and CRF in different ethnic populations of rural Kenya. A cross-sectional study was done among 1,099 rural Luo, Kamba, and Maasai of Kenya. Participants were 17-68 years old and 60.9% were women. Individual heart rate (HR) response to a submaximal steptest was used to assess CRF (estimated VO(2) max). Habitual PAEE was measured with combined accelerometry and HR monitoring, with individual calibration of HR using information from the step test. Men had higher PAEE than women (∼78 vs. ∼67 kJ day(-1) kg(-1) , respectively). CRF was similar in all three populations (∼38 and ∼43 mlO(2) ·kg(-1) min(-1) in women and men, respectively), while habitual PAEE measures were generally highest in the Maasai and Kamba. About 59% of time was spent sedentary (<1.5 METs), with Maasai women spending significantly less (55%). Both CRF and PAEE were lower in older compared to younger rural Kenyans, a difference which was most pronounced for PAEE in Maasai (-6.0 and -11.9 kJ day(-1) kg(-1) per 10-year age difference in women and men, respectively) and for CRF in Maasai men (-4.4 mlO(2) ·min(-1) kg(-1) per 10 years). Adjustment for hemoglobin did not materially change these associations. Physical activity levels among rural Kenyan adults are high, with highest levels observed in the Maasai and Kamba. The Kamba may be most resilient to age-related declines in physical activity.


Specific Secondary Bile Acids Control Chicken Necrotic Enteritis.
Necrotic enteritis (NE), mainly induced by the pathogens of Clostridium perfringens and coccidia, causes huge economic losses with limited intervention options in the poultry industry. This study investigated the role of specific bile acids on NE development. Day-old broiler chicks were assigned to six groups: noninfected, NE, and NE with four bile diets of 0.32% chicken bile, 0.15% commercial ox bile, 0.15% lithocholic acid (LCA), or 0.15% deoxycholic acid (DCA). The birds were infected with Eimeria maxima at day 18 and C. perfringens at day 23 and 24. The infected birds developed clinical NE signs. The NE birds suffered severe ileitis with villus blunting, crypt hyperplasia, epithelial line disintegration, and massive immune cell infiltration, while DCA and LCA prevented the ileitis histopathology. NE induced severe body weight gain (BWG) loss, while only DCA prevented NE-induced BWG loss. Notably, DCA reduced the NE-induced inflammatory response and the colonization and invasion of C. perfringens compared to NE birds. Consistently, NE reduced the total bile acids in the ileal digesta, while dietary DCA and commercial bile restored it. Together, this study showed that DCA and LCA reduced NE histopathology, suggesting that secondary bile acids, but not total bile acid levels, play an essential role in controlling the enteritis.


Effect of in vivo administration of anesthetics on GABAA receptor function.
In research involving tissue derived from animals, the use of anesthesia before sacrifice is recommended by the American Veterinary Medical Association and is strongly encouraged by university committees on the use of animals in research. In this study, the effects of anesthesia on the modulation of the GABAA receptor complex by ethanol, a benzodiazepine, and a barbiturate were determined. In vivo administration of the anesthetic methoxyflurane and CO2 before sacrifice resulted in a decrease in Cl- flux in mouse brain microsacs. These treatments also resulted in the loss of the ability of the GABAA receptor modulatory agents flunitrazepam and ethanol to enhance the Cl- flux in this assay system.


Screening-level risk assessment of methylmercury for non-anadromous Arctic char (Salvelinus alpinus).
Non-anadromous forms of Arctic char (Salvelinus alpinus), those that are restricted to lakes and rivers, typically have higher mercury (Hg) concentrations than anadromous forms, which migrate to and from the sea. Using tissue burden data from the literature and our own analyses, we performed a screening-level risk assessment of methylmercury (MeHg) for non-anadromous Arctic char. Our assessment included 1569 fish distributed across 83 sites. Site-specific mean total Hg concentrations in non-anadromous Arctic char muscle varied considerably from 0.01 to 1.13 µg/g wet weight, with 21% (17 of 83 sites) meeting or exceeding a threshold-effect level in fish of 0.33 µg/g wet weight, and 13% (11 of 83 sites) meeting or exceeding a threshold-effect level in fish of 0.5 µg/g wet weight. Of the sites in exceedance of the 0.33-µg/g threshold, 7 were located in Greenland and 10 in Canada (Labrador, Nunavut, and Yukon). All but one of these sites were located in interfrost or permafrost biomes. Maximum total Hg concentrations exceeded 0.33 µg/g wet weight at 53% of sites (40 of the 75 sites with available maximum Hg values), and exceeded 0.5 µg/g wet weight at 27% (20 of 75 sites). Collectively, these results indicate that certain populations of non-anadromous Arctic char located mainly in interfrost and permafrost regions may be at risk for MeHg toxicity. This approach provides a simple statistical assessment of MeHg risk to non-anadromous Arctic char, and does not indicate actual effects. We highlight the need for studies that evaluate the potential toxic effects of MeHg in non-anadromous Arctic char, as well as those that aid in the development of a MeHg toxic-effect threshold specific to this species of fish. Environ Toxicol Chem 2019;38:489-502. © 2018 SETAC.


Comparison of Tissue Yield Using Frictional Fabric Brush Versus Sharp Curettage For Endocervical Curettage.
The aim of the study was to evaluate and compare the diagnostic yield of conventional endocervical curettage (ECC) with fabric-based ECC in a consistent physician group. This is a retrospective case-control study of patients who underwent ECC both before and after introduction of a fabric-based ECC device. Histologic examination of curettings was categorized as satisfactory, limited, or inadequate. The Kruskall-Wallis test was used to compare proportions of gross descriptions and final diagnoses between groups. Between January 2010 and July 2011, 9234 ECCs were performed using conventional ECC technique. From September 2011 to October 2013, 774 ECCs were performed with the fabric-based ECC. Using the conventional ECC technique, 7809 (84.6%) of specimens were satisfactory, 1037 (11.2%) were limited, and 388 (4.2%) were inadequate and repeat biopsy was recommended. With fabric ECC, 705 (91.1%) of specimens were satisfactory, 64 (8.3%) were limited, and 5 (0.6%) were inadequate, and repeat biopsy was recommended. There were significantly fewer inadequate specimens with the fabric-based ECC (4.2% vs 0.6%, p < .001). Fabric-based ECC may significantly decrease inadequate and limited ECC specimens.


Ca L23 spectrum in amorphous and crystalline phases of calcium carbonate.
The calcium L(23) absorption edge is very sensitive to changes in local symmetry. Distinct spectra from calcite and aragonite as well as two different amorphous phases of calcium carbonate have been reported. Multiplet calculations using the CTM4XAS code, taking account of the local symmetry and the crystal field, suggest that the coordination octahedra are distorted by a change in length along one axis, the reduction in symmetry being coupled with tetrahedral displacements of the 4 atoms in the perpendicular plane. Examination of the polarization dependence implies that the two different amorphous phases arise from different levels of ordering of the oxygen coordination polyhedra around calcium. The change in the ordering suggests a pathway for the transformation to single crystal calcite. Although the code is unable to directly simulate the very low symmetry environment of Ca in aragonite, some degree of agreement is obtained with the same distortion of the coordination octahedron and greater tetrahedral displacements.


Internet Use and Psychological Well-Being at Advanced Age: Evidence from the English Longitudinal Study of Aging.
This work explores the connection between psychological well-being and Internet use in older adults. The study is based on a sample of 2314 participants in the English Longitudinal Study of Aging. The subjects, aged 50 years and older, were interviewed every two years over the 2006-2007 to 2014-2015 period. The connection between the use of Internet/Email and the main dimensions of psychological well-being (evaluative, hedonic and eudaimonic) was analyzed by means of three generalized estimating equation models that were fitted on 2-year lagged repeated measurements. The outcome variables, the scores on three well-being scales, were explained in terms of Internet/Email use, controlling for covariates that included health and socioeconomic indicators. The results support the existence of a direct relationship between Internet/Email use and psychological well-being. The connection between the main predictor and the score of the participants on the scale used to measure the eudaimonic aspect was positive and statistically significant at conventional levels (p-value: 0.015). However, the relevance of digital literacy on the evaluative and the hedonic components could not be confirmed (p-values for evaluative and hedonic dimensions were 0.078 and 0.192, respectively).


Power Optimized Smartwatch-Earbuds Multimodal System for Monitoring Activities of Daily Living.
Activities of daily living is an important entity to monitor for promoting healthy lifestyle for chronic disease patients, children and the healthy population. This paper presents a smartwatch and earbuds inertial sensors based multi-modal power efficient end-to-end mobile system for continuous, passive and accurate detection of broad daily activity classes. We collected various posture, stationary and moving activity data from 40 diverse subjects using earbuds and smartwatch and develop the novel power optimized end-to-end operational system consisting of i) optimized device sampling rates and Bluetooth packet transfer rates, ii) data buffering mechanism, iii) background services, and iv) optimized model size, and demonstrating 93% macro recall score in detecting various activities. Our power optimized solution uses 80%, 40% and 33.33% less battery power for the smartphone, smartwatch, and earbuds respectively, compared to a power agnostic system with an estimated continuous no-charging run time of 50 hours, 16.67 hours, and 25 hours for the smartphone, smartwatch, and earbuds respectively.Clinical relevance- The end-to-end power optimized activity detection system presented in this paper will assist practicing clinicians toward treatment of various chronic disease patients (e.g. diabetes, hypertension, heart disease and obesity) by long-term, continuous monitoring of their lifestyle and sedentary behavior.


Zoonotic parasites infecting free-living armadillos from Brazil.
Armadillos are specialist diggers and their burrows are used to find food, seek shelter and protect their pups. These burrows can also be shared with dozens of vertebrate and invertebrate species and; consequently, their parasites including the zoonotics. The aim of this study was to diagnose the presence of zoonotic parasites in four wild-caught armadillo species from two different Brazilian ecosystems, the Cerrado (Brazilian savanna) and the Pantanal (wetland). The investigated parasites and their correspondent diseases were: Toxoplasma gondii (toxoplasmosis), Trypanosoma cruzi (Chagas disease), Leishmania spp., (leishmaniasis), Paracoccidioides brasiliensis (Paracoccidioidomicosis) and Mycobacterium leprae (Hansen's disease). Forty-three free-living armadillos from Pantanal and seven road-killed armadillos from the Cerrado were sampled. Trypanosoma cruzi DTU TcIII were isolated from 2 out of 43 (4.65%) armadillos, including one of them also infected with Trypanosoma rangeli. Antibodies anti-T. gondii were detected in 13 out of 43 (30.2%) armadillos. All seven armadillos from Cerrado tested positive for P. brasiliensis DNA, in the lungs, spleen, liver fragments. Also, by molecular analysis, all 43 individuals were negative for M. leprae and Leishmania spp. Armadillos were infected by T. cruzi, T. rangeli, P. brasiliensis and presented seric antibodies to T. gondii, highlighting the importance of those armadillos could have in the epidemiology of zoonotic parasites.


Simple evolutionary pathways to complex proteins.
A recent paper in this journal has challenged the idea that complex adaptive features of proteins can be explained by known molecular, genetic, and evolutionary mechanisms. It is shown here that the conclusions of this prior work are an artifact of unwarranted biological assumptions, inappropriate mathematical modeling, and faulty logic. Numerous simple pathways exist by which adaptive multi-residue functions can evolve on time scales of a million years (or much less) in populations of only moderate size. Thus, the classical evolutionary trajectory of descent with modification is adequate to explain the diversification of protein functions.


Alcohol use and negative consequences among active duty military personnel.
An examination of alcohol use patterns in the active duty military to determine the relations of drinking levels and self-reported negative outcomes. A population-based cross-sectional study design using two-stage complex sampling methodology. Paper and pencil surveys were administered anonymously in groups at 64 U.S. military installations worldwide. Randomly selected active duty members (28,546) at major military installations representing the total active force, with the exception of recruits, cadets, and incarcerated personnel. Personnel were classified into five drinking levels ranging from abstainer to heavy drinker based on quantity and frequency of alcohol intake. Negative outcomes were measured as self-reported serious consequences of alcohol use and alcohol-related productivity loss. Risk for other alcohol related problems was assessed by the Alcohol Use Disorders Identification Test (AUDIT). Alcohol negative outcomes showed a curvilinear dose-response relationship with drinking levels. Higher levels of drinking were associated with higher rates of alcohol problems, but problem rates were notably higher for heavy drinkers. Heavy alcohol users showed nearly three times the rate of self-reported serious consequences and over twice the rate of self-reported productivity loss than moderate/heavy drinkers. Heavy drinkers also had the highest risk for alcohol problems on the AUDIT. One fifth of military personnel were heavy drinkers and were most likely aged 18 to 35. Prevention and clinical interventions should include a major focus on heavy drinkers. Commanders and peers should be trained in recognizing signs of heavy alcohol use and in approaching heavy alcohol users in a way that will foster positive attitudes as opposed to defensiveness and stigma.


Comparative susceptibility of mosquito populations in North Queensland, Australia to oral infection with dengue virus.
Dengue is the most prevalent arthropod-borne virus, with at least 40% of the world's population at risk of infection each year. In Australia, dengue is not endemic, but viremic travelers trigger outbreaks involving hundreds of cases. We compared the susceptibility of Aedes aegypti mosquitoes from two geographically isolated populations to two strains of dengue virus serotype 2. We found, interestingly, that mosquitoes from a city with no history of dengue were more susceptible to virus than mosquitoes from an outbreak-prone region, particularly with respect to one dengue strain. These findings suggest recent evolution of population-based differences in vector competence or different historical origins. Future genomic comparisons of these populations could reveal the genetic basis of vector competence and the relative role of selection and stochastic processes in shaping their differences. Lastly, we show the novel finding of a correlation between midgut dengue titer and titer in tissues colonized after dissemination.


G protein-independent G1 cell cycle block and apoptosis with morphine in adenocarcinoma cells: involvement of p53 phosphorylation.
Opioid effects on tumor growth have been a controversial topic of discussion. In the present study, morphine inhibited tumor cell proliferation at concentrations of >or=10 micro M. This was primarily caused by inhibition of cell cycle progression from G(1) to S phase. At higher concentrations (>or=500 micro M for 24 h), morphine also caused cell death. In nude mice, morphine significantly reduced the growth of MCF-7 and MDA-MB231 tumors but had no effect on HT-29 tumor growth. In these experiments, morphine plasma concentrations were similar to those found in cancer patients receiving chronic morphine treatment for pain relief (0.9-3.4 micro M). In MCF-7 and MDA-MB231 cells, morphine caused a naloxone (Nx)- and pertussis toxin-sensitive, concentration-dependent increase of GTPase activity, indicating that morphine signals could be transduced by opioid receptors via a G protein. However, the antiproliferative effects of morphine were not antagonized by Nx, pertussis toxin, forskolin, and 8-bromo-cAMP, suggesting that the typical opioid receptor-coupled signaling cascade involving the G(i), adenylyl cyclase, and protein kinase A was not involved. Instead, morphine caused an NH(2)-terminal phosphorylation of p53 at Ser(9) and/or Ser(15) and a stabilization of p53 in MCF-7 cells that express wild-type p53. p53 phosphorylation was not antagonized by Nx and resulted in an increase of p53-dependent proteins including p21, Bax, and the death receptor Fas. Blockade of Fas by Fas-fusion protein or inhibition of caspase 8 resulted in a partial inhibition of morphine-induced apoptosis. In addition, Fas ligand only induced apoptosis when administered together with morphine. However, the sensitivity of the tumor cells toward Fas ligand remained low. HT-29 cells, which express dominant negative p53 and show no increase of GTPase activity when treated with morphine, were less sensitive in vitro and were not affected in vivo. Our results suggest that morphine, alone or in combination with Nx, may reduce the growth of certain tumors, apparently in part through activation of p53.


A Biobank of Breast Cancer Explants with Preserved Intra-tumor Heterogeneity to Screen Anticancer Compounds.
The inter- and intra-tumor heterogeneity of breast cancer needs to be adequately captured in pre-clinical models. We have created a large collection of breast cancer patient-derived tumor xenografts (PDTXs), in which the morphological and molecular characteristics of the originating tumor are preserved through passaging in the mouse. An integrated platform combining in vivo maintenance of these PDTXs along with short-term cultures of PDTX-derived tumor cells (PDTCs) was optimized. Remarkably, the intra-tumor genomic clonal architecture present in the originating breast cancers was mostly preserved upon serial passaging in xenografts and in short-term cultured PDTCs. We assessed drug responses in PDTCs on a high-throughput platform and validated several ex vivo responses in vivo. The biobank represents a powerful resource for pre-clinical breast cancer pharmacogenomic studies (http://caldaslab.cruk.cam.ac.uk/bcape), including identification of biomarkers of response or resistance.


Activation of a peptidergic synapse locally modulates postsynaptic calcium influx.
We examined the synaptic connection between Phe-Met-Arg-Phe-NH2 (FMRFamide)-immunoreactive neurone VD4 and its target neurone P1, both found in the central nervous system of the pond snail Helisoma trivolvis. The major FMRFamide-like peak in neurone VD4 appears to be FMRFamide itself, based on its high performance liquid chromatography (HPLC) elution time and immunoreactivity before and after oxidation, but small peaks are also present at the elution times of Phe-Leu-Arg-Phe-NH2 (FLRFamide) and Gly-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (GDPFLRFamide). The modulatory actions of the neuropeptides found in neurone VD4 were tested on the postsynaptic target cell P1. Bath application of both the tetrapeptides FMRFamide and FLRFamide at a concentration of 10(-5) mol l-1 reduced the macroscopic voltage-sensitive calcium current of neurone P1 in culture; FMRFamide by 45% and FLRFamide by 51%. Bath application of the heptapeptide GDPFLRFamide (10(-5) mol l-1) reduced the calcium current by only 8%. We reconstructed the synaptic connection between VD4 and P1 in culture. Action-potential-evoked calcium transients in neurites growing from P1 cells in culture were monitored using Fura-2. Addition of FMRFamide, FLRFamide or GDPFLRFamide reduced the magnitude of the calcium transient in P1. Stimulation of VD4 mimicked the effects of peptide application and caused localized reductions in the action-potential-evoked calcium transients in P1 at the points of contact between the neurites of neurones VD4 and P1. These results suggest that neurone VD4 modulates the calcium influx of neurone P1 through the release of endogenous FMRFamide-related peptides and that this modulatory action is restricted to sites of synaptic interaction.


Survival, physiology, and lysis of Lactococcus lactis in the digestive tract.
The survival and the physiology of lactococcal cells in the different compartments of the digestive tracts of rats were studied in order to know better the fate of ingested lactic acid bacteria after oral administration. For this purpose, we used strains marked with reporter genes, the luxA-luxB gene of Vibrio harveyi and the gfp gene of Aequora victoria, that allowed us to differentiate the inoculated bacteria from food and the other intestinal bacteria. Luciferase was chosen to measure the metabolic activity of Lactococcus lactis in the digestive tract because it requires NADH, which is available only in metabolically active cells. The green fluorescent protein was used to assess the bacterial lysis independently of death. We report not only that specific factors affect the cell viability and integrity in some digestive tract compartments but also that the way bacteria are administrated has a dramatic impact. Lactococci which transit with the diet are quite resistant to gastric acidity (90 to 98% survival). In contrast, only 10 to 30% of bacteria survive in the duodenum. Viable cells are metabolically active in each compartment of the digestive tract, whereas most dead cells appear to be subject to rapid lysis. This property suggests that lactococci could be used as a vector to deliver specifically into the duodenum the proteins produced in the cytoplasm. This type of delivery vector would be particularly appropriate for targeting digestive enzymes such as lipase to treat pancreatic deficiencies.


Immune checkpoint inhibitors, endocrine adverse events, and outcomes of melanoma.
Immune checkpoint inhibitors (ICI) can cause endocrine adverse events. However, endocrine adverse events (AEs). However, endocrine AEs could be related to better treatment outcomes. Our aim was to investigate whether this holds true in a real-world setting of metastatic melanoma patients. A retrospective single-institution study. We included 140 consecutive metastatic melanoma patients treated with ICI between January 2012 and May 2019. We assessed the endocrine toxicity and the best possible treatment outcomes from electronic patient records, including laboratory parameters and radiological images. Of the treated patients, 21 patients (15%) were treated with ipilimumab, 46 (33%) with nivolumab, 67 (48%) with pembrolizumab, and 6 (4%) with combination therapy (ipilimumab + nivolumab). Endocrine AEs appeared in 29% (41/140) patients. Three patients had two different endocrine AEs. Thyroid disorders were the most common: 26% (36/140), followed by hypophysitis: 4% (5/140). Three subjects (2%, 3/140) were diagnosed with autoimmune diabetes. Three patients had to terminate treatment due to endocrine toxicity. Radiological manifestations of endocrine AEs were found in 16 patients (39%, 16/41). Endocrine toxicity was associated with significantly better treatment outcomes. Median progression-free survival (8.1 months, range 5.1-11.1 months vs 2.7 months, range 2.4-3.0 months, P < 0.001), and median overall survival (47.5 months, range 15.5-79.5 months vs 23.7 months, range 15.3-32.1 months, P = 0.035) were longer for patients experiencing endocrine AEs. The higher number of endocrine AEs suggest that regular laboratory monitoring aids in AE detection. Endocrine AEs in metastatic melanoma may correlate with better treatment outcomes.


Three-dimensional regional dynamics of the normal mitral anulus during left ventricular ejection.
The mitral anulus is a dynamic structure that undergoes alterations in size and shape throughout the cardiac cycle, contracting during systole. Numerous reports have shown this systolic orifice reduction to be due chiefly to posterior annular contraction, whereas the anterior perimeter was unchanged. Segmental motion of the mitral anulus from true in vivo three-dimensional data, however, has not been described. We used radiopaque markers and simultaneous biplane videofluoroscopy to measure the lengths of mitral anular segments in seven closed-chest, sedated dogs. Eight markers were placed equidistant from each other around the mitral anulus, As viewed from the left atrium, segment 1 began at the posteromedial commissure, and the remaining segments were numbered sequentially clockwise around the anulus (that is, the posterior mitral anulus encompassed segments 1 to 4 and the anterior anulus encompassed segments 5 to 8). Marker image coordinates obtained from two orthogonal views 7 to 12 days after implantation were merged to construct three-dimensional marker coordinates at end-diastole and end-systole. From end-diastole to end-systole, mean annular area decreased by 11% +/- 8% (5.5 +/- 0.9 cm2 to 4.9 +/- 0.8 cm2, p = 0.005) and perimeter by 5% +/- 4% (8.8 +/- 0.7 cm to 8.3 +/- 0.7 cm, p < 0.01). Mitral annular segmental percent systolic shortening (negative values indicate lengthening) were as follows (mean +/- standard deviation): segment 1, 7% +/- 9%; segment 2, 8% +/- 10%; segment 3, 16% +/- 6%; segment 4, 10% +/- 7%; segment 5, -4% +/- 5%, segment 6, -7% +/-7%; segment 7, 3% +/- 2%; and segment 8, 6% +/- 5%. With the exception of segment 1, all posterior (2 to 4) and two anterior (7 and 8) mitral annular segments contracted significantly (p < or = vs zero, paired t test). Two anterior annular segments (5 and 6, regions overlapping aortic-mitral continuity), however, unexpectedly lengthened during left ventricular systole. We conclude that the anterior mitral anulus may be a much more dynamic component of the mitral apparatus that previously thought. Such heterogeneous dynamic annular motion should be taken into account when various mitral valve reparative techniques are being designed.


Projections from the entorhinal cortex, perirhinal cortex, presubiculum, and parasubiculum to the medial thalamus in macaque monkeys: identifying different pathways using disconnection techniques.
The projections from the perirhinal cortex, entorhinal cortex, parasubiculum, and presubiculum to the thalamus were examined using both anterograde and retrograde tracers. Attention focused on the routes taken by these projections, which were delineated by combining surgical tract section with the placement of a tracer. Projections to the anterior thalamic nuclei almost exclusively used the fornix. These relatively light projections, which arose from all areas of the entorhinal cortex, from the presubiculum, parasubiculum, and area 35 of the perirhinal cortex, terminated mainly in the anterior ventral nucleus. In contrast, the projections to the lateral dorsal nucleus from the entorhinal cortex, presubiculum and parasubiculum were denser than those to the anterior thalamic nuclei. The projections to the lateral dorsal nucleus used two routes. While nearly all of the projections from the subicular complex used the fornix, many of the entorhinal cortex projections passed caudally in the temporopulvinar bundle to reach the lateral dorsal nucleus. The perirhinal cortex, as well as the entorhinal cortex, also projects to nucleus medialis dorsalis. These projections exclusively used the external capsule and thence the inferior thalamic peduncle. Other temporal-thalamic projections included those to the medial pulvinar, via the temporopulvinar bundle, from the perirhinal and entorhinal cortices, and those to the paraventricular nucleus from the entorhinal cortex. By identifying these routes, it is possible to appreciate how different lesions might disconnect temporal-diencephalic pathways and so contribute to memory disorders.


Esterase electrophoretic pattern relatedness between Shigella species and Escherichia coli.
Esterases of 57 strains of Shigella dysenteriae, Sh. flexneri, Sh. boydii and Sh. sonnei and 26 strains of Escherichia coli, including the Alkalescens Dispar group, were compared by polyacrylamide-agarose gel electrophoresis. Six types of esterase bands differing in their ability to hydrolyse synthetic substrates and in their sensitivity to heat and to di-isofluoropropyl phosphate were defined. Individual activities and sensitivities of these bands and the apparent molecular weight of the major esterase, estimated to be 58000 by polyacrylamide gradient gel electrophoresis, were identical for both Shigella species and E. coli. One esterase with a molecular weight of 104000 was found in some strains of E. coli. Variations in the number and mobility of bands among Shigella strains defined different esterase patterns (zymotypes) which appeared to be distinct for each species.


Antiinflammatory activity of alpha-methyl-3-phenyl-7-benzofuranacetic acid (R-803).
The antiinflammatory activity of R-803 (alpha-methyl-3-phenyl-7-benzofuranacetic acid), a new nonsteroidal drug, has been demonstrated. The oral administration of R-803 inhibits carrageenan-induced edema of the rat's paw, ultraviolet-induced erythema of guinea-pig skin, adjuvant-induced and 6-sulfonamidoindazole-induced arthritides of the rat at doses in the range of 1.0 to 6.0 mg/kg. It also inhibits the cotton pellet-induced granuloma in the rat at higher doses.


Responsiveness of the foot and ankle ability measure (FAAM) in individuals with diabetes.
The impact of diabetes on physical function pose a challenge in assessing clinical outcomes. The purpose of this study was to provide evidence of responsiveness for the foot and ankle ability measures (FAAM) in individuals with diabetes mellitus. The two most recent FAAM scores of 155 diabetic patients treated for foot/ankle pathology were analyzed. Based on physical component summary (PCS) scores of the SF-36, subjects were categorized as improved (>7-point positive change), worsened (>7-point negative change), or unchanged (<7-point change). Analyses of the worsened and improved groups were compared to the unchanged group using two-way repeated measures ANOVAs and ROC curve analyses. The ANOVAs demonstrated a significant difference between groups (P = 0.001). ROC curves analysis for detecting an improvement or decline in status were 0.73 (95% CI 0.62-0.84) and 0.70 (95% CI 0.59-0.81), respectively. An increase in FAAM score of 9 points represented the minimal clinically important difference (MCID) with 0.64 sensitivity and 0.78 specificity. A decrease in FAAM score of 2 points represented a MCID with 0.65 sensitivity and 0.61 specificity. The FAAM demonstrated responsiveness to change in individuals with orthopedic foot and ankle dysfunction complicated by diabetes and can be used to measure patient outcomes over a 6-month period.


Metals and DNA: molecular left-handed complements.
Chiral metal complexes provide unique molecular probes for DNA. Chiral reagents that "recognize" different local structures along the DNA strand have been designed by a process in which the asymmetry in shape and size of the complex is matched to that of the DNA helical groove. As a result, the chiral metal complexes provide very sensitive probes for local helical structure, both left- and right-handed. Direct coordination of chiral complexes to the DNA bases adds an element of sequence selectivity to the probe design. With a suitable reactive metal center, reagents that target chemically specific sites along the strand may be developed. One such chiral reagent, which cleaves left-handed DNA sites with photoactivation, has been useful in mapping this distinct conformation and examining its biological role. The conformation-specific molecular cleaver, much like a DNA-binding enzyme, recognizes and reacts at discrete sites along the DNA strand. These site-specific chiral metal complexes provide exciting new tools for probing the local variations in DNA structure and its role in the regulation of gene expression.


The effect of different posts on fracture strength of roots with vertical fracture and re-attached fragments.
The aim of this in vitro study was to test the effect of different post systems on fracture strength of roots with re-attached fragments. Root canals of eighty extracted single-rooted human teeth were instrumented (ProFile) and randomly divided into two groups. The roots in the first group were vertically cracked, and the fragments were re-attached using Super Bond C&B (Sun Medical, Tokya, Japan). The roots in the second group were kept sound. Obturation of the roots was performed with MetaSEAL (Sun Medical) and gutta-percha. Post spaces were prepared, and the roots were restored with one of the followings: UniCore (Ultradent), Everstick (Stick Tech), Ribbond (Ribbond), ParaPost (Coltene/Whaledent) (n = 10). Four mm high build-ups were created (Clearfil DC Bond Core; Kuraray, Tokyo, Japan). Compressive loading of the samples was performed after 24 h (1 mm min(-1)). Mean load necessary to fracture each sample was recorded (Newton) and statistically analysed (One-way anova, t-tests). ParaPost showed the highest fracture strength among the roots with re-attached fragments (P < 0.05). UniCore and ParaPost systems showed similar fracture strength in the sound roots (P > 0.05). Re-attached fragments significantly reduced the fracture strength of roots in UniCore group (P = 0.000). Ribbond post showed mostly repairable fractures. Metal post (ParaPost) showed the highest fracture strength in the roots with re-attached fragments; however, fracture pattern was 41% non-repairable. Re-attached fragments significantly reduced the fracture strength of the roots in UniCore group. Prefabricated posts showed similar fracture strength in the sound roots. Customized post systems EverStick and Ribbond showed mostly repairable failure after loading in sound roots or roots with re-attached fragments.


The effect of amniotic membrane extract on umbilical cord blood mesenchymal stem cell expansion: is there any need to save the amniotic membrane besides the umbilical cord blood?
Umbilical cord blood is a good source of the mesenchymal stem cells that can be banked, expanded and used in regenerative medicine. The objective of this study was to test whether amniotic membrane extract, as a rich source of growth factors such as basic-fibroblast growth factor, can promote the proliferation potential of the umbilical cord mesenchymal stem cells. The study design was interventional. Umbilical cord mesenchymal stem cells were isolated from voluntary healthy infants from hospitals in Shiraz, Iran, cultured in the presence of basic-fibroblast growth factor and amniotic membrane extracts (from pooled - samples), and compared with control cultures. Proliferation assay was performed and duplication number and time were calculated. The expression of stem cell's specific markers and the differentiation capacity toward osteogenic and adipogenic lineages were evaluated. Amniotic membrane extract led to a significant increase in the proliferation rate and duplication number and a decrease in the duplication time without any change in the cell morphology. Both amniotic membrane extract and basic-fibroblast growth factor altered the expressing of CD44 and CD105 in cell population. Treating basic-fibroblast growth factor but not the amniotic membrane extract favored the differentiation potential of the stem cells toward osteogenic lineage. The amniotic membrane extract administration accelerated cell proliferation and modified the CD marker characteristics which may be due to the induction of differentiation toward a specific lineage. Amniotic membrane extract may enhance the proliferation rate and duplication number of the stem cell through changing the duplication time.


Histopathological changes induced by extracts from the tissue covering the stingers of Potamotrygon falkneri freshwater stingrays.
Pain is the most conspicuous symptom observed in patients wounded by stingrays, and skin necrosis is common in accidents by freshwater stingrays. The extract from the stinger integumentary tissue of Potamotrygon falkneri containing toxic components (venom) was tested for its ability to induce histopathological changes in the dorsal skin of mice at different times. 3-6 h after injection, foci of necrosis in isolated basal epidermal cells were observed. Full coagulative necrosis of the skin, subcutaneous tissue and skeletal muscle was evident as soon as 24 h after venom exposure, with a clear demarcation from the normal skin. After 48 h, round collections of necrotic cells start to coalesce originating extensive skin necrotic plaques that detach from viable tissue after 72-96 h. Inflammatory infiltrate was observed after 6 h, but was always mild. Acute vascular thrombosis was rare, and hemorrhage was not present at any time. Superficial bacterial infection was present in two of the examined cases. In conclusion, the venom of P. falkneri is responsible for the development of an early necrosis with mild inflammatory reaction, probably due to direct action of the venom. The severe local damage is probably worsened by the mechanical trauma caused by the stinger.


[Measles].
In April 2019, UNICEF denounced that more than 20 million children worldwide had not been vaccinated and alerted on possible outbreaks of measles which, due to the high transmissibility of this virus, is the first disease preventable by vaccination to emerge. If the decline in vaccinations continues, pertussis, tetanus and other diseases, which require less coverage to achieve population protection, may also reappear. In Argentina, the current outbreak began in late August 2019. Measles virus is transmitted by air, infects multiple organs, and is associated with immunosuppression. Its genome consists of single stranded RNA. Genotyping is carried out by sequencing a 450-nucleotide fragment of the N protein, which contains the highest density of nucleotide variation. In South America, D8 is the circulating genotype and in North America, B3 accounts for 8% of the cases. Each person with measles infects, on average, another 12-18 people in a susceptible population. Vaccination confers direct and indirect protection, and induces both antibodies and cellular immunity. Newborns are protected by maternal antibodies transmitted via the placenta, up to 6 months. In Argentina, the Vaccination Calendar includes two doses of triple viral vaccine, at 12 months and 5 years, and a zero dose (6-11 months of age) in districts with disease cases. The protection conferred by the vaccine is 93% at 12 months with a dose, and with 2 doses 97% for life.


A genome-wide association study for clinical mastitis in first parity US Holstein cows using single-step approach and genomic matrix re-weighting procedure.
Clinical mastitis (CM) is one of the health disorders with large impacts on dairy farming profitability and animal welfare. The objective of this study was to perform a genome-wide association study (GWAS) for CM in first-lactation Holstein. Producer-recorded mastitis event information for 103,585 first-lactation cows were used, together with genotype information on 1,361 bulls from the Illumina BovineSNP50 BeadChip. Single-step genomic-BLUP methodology was used to incorporate genomic data into a threshold-liability model. Association analysis confirmed that CM follows a highly polygenic mode of inheritance. However, 10-adjacent-SNP windows showed that regions on chromosomes 2, 14 and 20 have impacts on genetic variation for CM. Some of the genes located on chromosome 14 (LY6K, LY6D, LYNX1, LYPD2, SLURP1, PSCA) are part of the lymphocyte-antigen-6 complex (LY6) known for its neutrophil regulation function linked to the major histocompatibility complex. Other genes on chromosome 2 were also involved in regulating immune response (IFIH1, LY75, and DPP4), or are themselves regulated in the presence of specific pathogens (ITGB6, NR4A2). Other genes annotated on chromosome 20 are involved in mammary gland metabolism (GHR, OXCT1), antibody production and phagocytosis of bacterial cells (C6, C7, C9, C1QTNF3), tumor suppression (DAB2), involution of mammary epithelium (OSMR) and cytokine regulation (PRLR). DAVID enrichment analysis revealed 5 KEGG pathways. The JAK-STAT signaling pathway (cell proliferation and apoptosis) and the 'Cytokine-cytokine receptor interaction' (cytokine and interleukines response to infectious agents) are co-regulated and linked to the 'ABC transporters' pathway also found here. Gene network analysis performed using GeneMania revealed a co-expression network where 665 interactions existed among 145 of the genes reported above. Clinical mastitis is a complex trait and the different genes regulating immune response are known to be pathogen-specific. Despite the lack of information in this study, candidate QTL for CM were identified in the US Holstein population.


We're just about average. Putting New York State residents' health and social conditions in perspective.
A series of reports on health and social standards place New York State residents at a mid position in comparison to national statistics and data for other states. The question is raised, should we be satisfied with being "just average"?


Tamoxifen enhances choline acetyltransferase mRNA expression in rat basal forebrain cholinergic neurons.
Novel estrogen-like molecules known as SERMs (selective estrogen receptor modulators) produce many of the beneficial estrogen-like actions without the detrimental side-effects. The SERM, tamoxifen, an estrogen-like molecule with both agonist and antagonist properties, is widely prescribed for the treatment of breast cancer. While the effects of tamoxifen are being evaluated in many peripheral tissues, its effects in the central nervous system (CNS) have been largely ignored. In the present study, we begin to evaluate the effects of tamoxifen in the rat basal forebrain, a region known to be highly responsive to estrogen. We compared the effects of short-term (24 h) tamoxifen treatment to that of estrogen on ChAT mRNA expression in cholinergic neurons. In addition, we examined the effect of tamoxifen in the presence and absence of estrogen. Our results indicate that tamoxifen enhances ChAT expression in a manner similar to that of estrogen in several basal forebrain regions. In contrast, tamoxifen exhibits antagonist properties with respect to estrogen-induction of progesterone receptor mRNA in the medial preoptic nucleus. These results indicate tamoxifen has estrogenic properties with respect to cholinergic neurons, suggesting a previously unidentified effect of this agent in the CNS.


Corticosteroid tapering with benralizumab treatment for eosinophilic asthma: PONENTE Trial.
Benralizumab is an interleukin-5 receptor α-directed cytolytic monoclonal antibody approved in several countries for the add-on maintenance treatment of patients with severe eosinophilic asthma aged 12 years and older. In the 28-week Phase III ZONDA trial (ClinicalTrials.gov identifier: NCT02075255), benralizumab produced a median 75% reduction from baseline in oral corticosteroid (OCS) dosage (versus 25% for placebo) while maintaining asthma control for patients with OCS-dependent severe asthma. This manuscript presents the detailed protocol for the Phase IIIb PONENTE (ClinicalTrials.gov identifier: NCT03557307), a study that will build on the findings from ZONDA. As the largest steroid-sparing study undertaken in severe asthma, PONENTE has a faster steroid tapering schedule for prednisone dosages ≥7.5 mg·day-1 than previous studies, and it includes an evaluation of adrenal insufficiency and an algorithm to taper OCS dosage when prednisone dosage is ≤5 mg·day-1. It also has a longer maintenance phase to assess asthma control for up to 6 months after completion of OCS tapering. The two primary endpoints are whether patients achieve 100% reduction in daily OCS use and whether patients achieve 100% reduction in daily OCS or achieve OCS dosage ≤5 mg·day-1, if adrenal insufficiency prevented further reduction, both sustained over ≥4 weeks without worsening of asthma. Safety and change from baseline in health-related quality of life will also be assessed. PONENTE should provide valuable guidance for clinicians on tapering OCS dosage, including the management of adrenal insufficiency, following benralizumab initiation for the treatment of patients who are OCS-dependent with severe, uncontrolled eosinophilic asthma.


Quad-trees, oct-trees, and k-trees: a generalized approach to recursive decomposition of euclidean space.
K-trees are developed as a K-dimensional analog of quad-trees and oct-trees. K-trees can be used for modeling K-dimensional data. A fast algorithm is given for finding the boundary size of a K-dimensional object represented by a K-tree. For K considered as con-stant; the algorithm provides a method for computing the perimeter of a quad-tree encoded image or the surface area of an oct-tree encoded object in worst case time proportional to the number of nodes in the tree. This improves upon the expected-case linear-time method of Samet [10] for the perimeter problem. Our method has been implemented in Pascal, and a computational example is given.


Age influences on the molecular presentation of tumours.
Cancer is often called a disease of aging. There are numerous ways in which cancer epidemiology and behaviour change with the age of the patient. The molecular bases for these relationships remain largely underexplored. To characterise them, we analyse age-associations in the nuclear and mitochondrial somatic mutational landscape of 20,033 tumours across 35 tumour-types. Age influences both the number of mutations in a tumour (0.077 mutations per megabase per year) and their evolutionary timing. Specific mutational signatures are associated with age, reflecting differences in exogenous and endogenous oncogenic processes such as a greater influence of tobacco use in the tumours of younger patients, but higher activity of DNA damage repair signatures in those of older patients. We find that known cancer driver genes such as CDKN2A and CREBBP are mutated in age-associated frequencies, and these alter the transcriptome and predict for clinical outcomes. These effects are most striking in brain cancers where alterations like SUFU loss and ATRX mutation are age-dependent prognostic biomarkers. Using three cancer datasets, we show that age shapes the somatic mutational landscape of cancer, with clinical implications.


Factors affecting the developmental competence of mouse oocytes grown in vitro: oxygen concentration.
The purpose of this study was to assess the effects of oxygen concentration on the developmental competence of mouse oocytes grown in vitro because oxygen has been shown to affect the nuclear maturation of oocytes (Haidri et al., 1971: J Reprod Fertil 26:409-411) and preimplantation embryo development (Whitten, 1971: Adv Biosci 6:129-139). Oocyte--granulosa cell complexes were isolated from the preantral follicles of 12-day-old mice and cultured for 10 days in serum-free medium equilibrated with 5, 10, 15, or 20% O2. Five percent CO2 was used for all groups. Oocytes from all groups were then matured and fertilized, and preimplantation embryos cultured using 5% O2. With increased oxygen tension, there were dramatic decreases in the percentage of (1) oocytes that survived in vitro culture, (2) surviving oocytes that could resume meiosis and undergo germinal vesicle breakdown (GVB), and then cleave to the two-cell stage, and (3) two-cell-stage embryos that completed the blastocyst transition. For example, 42% of the oocytes grown in a 5% O2 atmosphere cleaved to the two-cell stage compared with only 3% when the oocytes grew in an atmosphere of 20% O2. These dramatic effects of elevated oxygen were mitigated by either increased numbers of the oocyte-granulosa cell complexes in the culture or increased concentration of oxygen late in the culture period, after the oocytes became surrounded by greater numbers of granulosa cells. When the culture period was extended for 4 days beyond the standard 10 day cultures used in the experiments described above, there was an increased occurrence of precocious GVB or failure of the somatic cells to maintain meiotic arrest. This was prevented by increased oxygen concentration. Nevertheless, extending the time of culture even in the presence of elevated oxygen failed to increase oocyte growth to the equivalent of in vivo-grown oocytes or to improve the developmental competence of the in vitro-grown oocytes. It was concluded that concentrations of O2 above 5% have a deleterious effect on oocyte development during the early stages of culture. However, increasing the concentration on O2 during the later stages, in conjunction with other treatments to promote normal granulosa cell development and function and their interaction with the oocyte, may be critical to promote normal oocyte development in vitro.


Purification of TEM-1 beta-lactamase by immunoaffinity chromatography.
A monoclonal antibody prepared against TEM-1 beta-lactamase was found to compete with penicillins and cephalosporins for binding to the enzyme. The purified antibody preparation was linked to Sepharose 4B and used for immunoaffinity-chromatography purification of TEM-1 beta-lactamase. Elution with either benzylpenicillin or cloxacillin yielded a highly purified, concentrated and active enzyme preparation.


Dietary intake, leisure time activities and obesity among adolescents in Western Sweden: a cross-sectional study.
Overweight and obesity among adolescents are increasing worldwide. Risk factors include dietary intake characteristics and high levels of physical inactivity. In Sweden, few large comprehensive population-based surveys of dietary intake and lifestyle among adolescents have been carried out. Thus, the purpose of the current study was to describe dietary intake and food choices as well as leisure time activities in relation to overweight and obesity in a total sample of all schoolchildren aged 15 years in Western Sweden. In 2008, a questionnaire was sent to all 21,651 adolescents born in 1992 in Västra Götaland Region, Sweden. Participation rate was 54.3 % (50.7 % girls/49.3 % boys). The questionnaire included a 73-item semi-quantitative food frequency questionnaire and questions on lifestyle. Results were evaluated against the Nordic Nutrition Recommendations and Swedish indicators of healthy diet and exercise habits. Associations with concurrent overweight and obesity were evaluated in multiple linear regression analysis. Among girls, 49.5 % reached the goal of consuming fruit and vegetables at least daily, whereas for boys the figure was 34.4 %. Among both sexes, 15 % reached the goal of consuming fish at least twice weekly. Two-thirds of both sexes reached the goal of regular moderate or vigorous physical activity weekly. In total, 12.4 % were overweight and 2.4 % were obese. More girls than boys were underweight, whereas more boys than girls were overweight or obese (p < 0.001). Boys exhibited a more frequent intake of sodas and concentrated fruit juices, milk 3 % fat, bread and potatoes and fast food (p < 0.001). Frequent intake of candies and chocolate was reported by both sexes. Among girls and boys, living in rural areas, living in apartments and reporting no frequent leisure time physical activity were significant risk factors for being overweight or obese, also when adjusted for other risk factors. Dietary habits of adolescents in Western Sweden warrant improvements. Public health actions should be taken to increase consumption of fruit, vegetables and fish, and decrease consumption of sodas and candies and also to increase frequency of physical activity. These actions may be helpful in reducing risks for overweight and obesity.


Impact of fibroblast growth factor-2 on tumor microvascular architecture. A tridimensional morphometric study.
Three cell clones originated by transfection of human endometrial adenocarcinoma HEC-1-B cells with fibroblast growth factor-2 (FGF-2) cDNA and characterized by a different capacity to produce and secrete the growth factor were transplanted subcutaneously in nude mice. Corrosion casting of the tumor microvasculature of xenografts produced by injection of 2 x 10(6) or 10 x 10(6) FGF-2-B9 cells (which produce and secrete significant amounts of FGF-2), 10 x 10(6) FGF-2-A8 cells (which produce comparable amounts of FGF-2 but do not secrete it), or 10 x 10(6) control FGF-2-B8 cells (which produce only trace amounts of FGF-2) was performed after 14 days of growth. Interbranching distances, intervascular distances, branching angles, and vessel diameters were then determined using tridimensional stereo pairs of the casted tumor vascularity. When transplanted at the same concentration, FGF-2-B9 cells grew faster in nude mice compared with FGF-2-A8 and FGF-2-B8 clones. The total amount of new vessel formation was far higher in FGF-2-B9 tumors than in FGF-2-B8 or FGF-2-A8 tumors. Also, vessel courses were more irregular and blind-ending vessels and evasates were more frequent in FGF-2-B9 tumors. Moreover, FGF-2-B9 tumor microvasculature was characterized by a wider average vascular diameter and by an extreme variability of the diameter of each individual vessel along its course between two ramifications. No statistical differences were observed when the distribution curves of the values of intervascular distances, interbranching distances, and branching angles of the microvessel network were compared among the different experimental groups. The distinctive features of the microvasculature of FGF-2-B9 tumors were retained, at least in part, in the smaller lesions produced by injection of a limited number of cells. The data indicate that FGF-2 production and release confer to FGF-2-B9 cells the ability to stimulate the formation of new blood vessels with distinctive architectural features. Neovascularization of FGF-2-B9 lesions parallels the faster rate of growth of the neoplastic parenchyma. This does not affect the overall architecture of the microvessel network that appears to be primed by characteristics of the HEC-1-B tumor cell line and/or by the microenvironment of the host. To our knowledge, this work represents the first attempt to define the influence of a single, defined growth factor on the tridimensional tumor vascular pattern.


Intrathecal triiodothyronine administration causes greater heart rate stimulation in hypothyroid rats than intravenously delivered hormone. Evidence for a central nervous system site of thyroid hormone action.
To determine whether intracerebrally localized iodothyronines produce thyroid hormone-related functional effects, heart rate responses were compared in conscious hypothyroid rats given triiodothyronine (T3) by either the intrathecal or the intravenous route. A significant increase in heart rate occurred within 18 h after 1.5 nmol T3/100 g body wt was delivered intrathecally through a cannula previously placed in the lateral cerebral ventricle. Injection of the same T3 dose intravenously through an indwelling jugular catheter or injection of vehicle only by either route produced no significant increase in heart rate during the 48-h postinjection period of observation. These differences were observed even though integrated serum T3 concentrations were significantly lower after intrathecal than after intravenous T3 injection. The results indicate that thyroid hormone effects on heart rate are exerted within the brain as well as within the heart.


[Development of medical laboratory equipment for mass screening of the population].
Mass health screening dictates the necessity to expand the range and output of clinical laboratory devices in medical industrial production, to create and introduce devices for microanalysis in clinical practice. The successful realization of the above problems calls for an integrated approach to develop engineering methodological, chemical and metrologic provision of the clinical and diagnostic laboratory studies, to solve technological and organizational problems in developing and producing the new generation of clinical laboratory devices and sets.


Association between circulating miRNAs and spinal involvement in patients with axial spondyloarthritis.
Dysregulation of miRNAs and their target genes contributes to the pathophysiology of autoimmune diseases. Circulating miRNAs may serve as diagnostic/prognostic biomarkers. We aimed to investigate the association between circulating miRNAs, disease activity and spinal involvement in patients with axial spondyloarthritis (AxSpA). Total RNA was isolated from the plasma of patients with non-radiographic (nr)AxSpA, patients with ankylosing spondylitis (AS) and healthy controls (HC) via phenol-chloroform extraction. A total of 760 miRNAs were analysed with TaqMan® Low Density Arrays, and the expression of 21 miRNAs was assessed using single assays. Comprehensive analysis demonstrated the differential expression of miRNAs among patients with progressive spinal disease. Of the 21 miRNAs selected according to their expression patterns, the levels of miR-625-3p were significantly different between nr-AxSpA patients and HCs. We found no correlation between miRNA levels and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) in nr-AxSpA patients. Selected miRNAs, such as miR-29a-3p, miR-146a-5p or miR-222-3p with an established role in extracellular matrix formation and inflammation were associated with spinal changes and/or disease activity assessed by BASDAI in AS patients, including miR-625-3p reflecting disease activity in AS with spinal involvement. Our data indicate that circulating miRNAs play a role in the pathogenesis of AxSpA and are also suggestive of their potential as biomarkers of disease progression. We hypothesize that differential systemic levels of miRNA expression reflect miRNA dysregulation at sites of spinal inflammation or bone formation where these molecules contribute to the development of pathophysiological features typical of AxSpA.


Enhanced Electrical Properties of Lithography-Free Fabricated MoS2 Field Effect Transistors with Chromium Contacts.
Molybdenum disulfide (MoS2) as a two-dimensional semiconductor material has been actively explored for field-effect-transistors (FETs). The current prevailing method for MoS2 FET fabrication involves multiple complex steps, including electron beam (e-beam) lithography, annealing, etc., which are time-consuming and require polymer resists. As a consequence, the MoS2 exposed to chemicals during the patterning process may be unfavorably affected by residues and the performance of the final FET could be impaired while the annealing limits materials for FETs. Therefore, there is an urgent need to free the fabrication of FETs from e-beam lithography and annealing. In this study, we introduce an e-beam lithography-free method to fabricate MoS2 FETs by employing maze-like source/drain electrodes. In addition, an ohmic contact in multilayer MoS2 FETs using chromium (Cr) as source/drain electrodes is achieved without annealing. The underlying mechanism for contact performance is studied, and the tightness of the contact and the type of metal are found to be responsible because they determine the contact resistance. Furthermore, the long-term device degradation is explored, in which the oxidation of metal dominates. The facile fabrication process and mechanism explanation in this work might provide a new platform for future electronic devices.


A "stealth effect": adenocarcinoma cells engineered to express TRAIL elude tumor-specific and allogeneic T cell reactions.
BALB/c mammary adenocarcinoma cells engineered to express TNF-related apoptosis-inducing ligand (TRAIL)/APO-2 ligand (APO-2L) on their membrane (TSA-TRAIL) grow with kinetics similar to that of parental cells (TSA-pc) in vitro and in nu/nu mice. In contrast, TSA-TRAIL cells grow faster than TSA-pc in normal BALB/c mice. In DBA/2 mice, which differ from BALB/c mice at minor histocompatibility Ags, they also grow faster and display a higher percentage of tumor takes than TSA-pc. In fully histoincompatible C57BL/6 (B6) mice, TSA-TRAIL cells form evident tumors that are slowly rejected by most mice, but outgrow in a few. In contrast, TSA-pc cells are rejected at once by B6 mice. Since TRAIL/APO-2L induces apoptosis by interacting with a variety of specific receptors, this rapid growth in both syngeneic and allogeneic mice may be the result of an immunosuppressive mechanism. The following evidence supports this hypothesis: 1) TSA-TRAIL cells overcome the strong immunity against TSA-pc cells elicited in BALB/c mice by preimmunization with TSA cells engineered to release IL-4; 2) their rejection by B6 mice does not prime a CTL-mediated memory; 3) thymidine uptake by T lymphocytes unstimulated or stimulated by allogeneic cells is inhibited when TSA-TRAIL cells are added as third party cells; 4) CTL kill TSA-pc but not TSA-TRAIL cells in 48-h assays; and 5) activated lymphocytes interacting with TSA-TRAIL cells in vivo and in vitro undergo apoptosis.


DNA polymerase delta: gene sequences from Plasmodium falciparum indicate that this enzyme is more highly conserved than DNA polymerase alpha.
Genes encoding proteins homologous to the catalytic subunits of DNA polymerase alpha and delta have been cloned from the human malaria parasite Plasmodium falciparum. These are among the first cellular replicative DNA polymerase genes to be cloned and their sequences allow us to make new statements about the relative degrees of conservation of these two enzymes. The most important finding was that P. falciparum Pol delta showed considerable homology to the only other Pol delta enzyme for which published sequence is available, that of S. cerevisiae, displaying an overall amino acid identity of 45% and identity over a highly conserved central region of 59%. In contrast, the level of identity shown over the equivalent central region of Pol alpha between the P. falciparum and S. cerevisiae sequences is only 32%. The sequence data also allowed us to examine the degree of conservation in putative exonuclease domains of Pol delta. The Pol delta gene of P. falciparum maps to chromosome 10 and evidence is presented for the presence of different sized Pol delta mRNA's in the asexual and sexual erythrocytic stages of parasite development.


Sulfatase-1 overexpression indicates poor prognosis in urothelial carcinoma of the urinary bladder and upper tract.
Urothelial carcinoma (UC), arising from the urothelium of the urinary tract, can occur in the upper (UTUC) and the urinary bladder (UBUC). A representative molecular aberration for UC characteristics and prognosis remains unclear. Data mining of Gene Expression Omnibus focusing on UBUC, we identified sulfatase-1 (SULF1) upregulation is associated with UC progression. SULF1 controls the sulfation status of heparan sulfate proteoglycans and plays a role in tumor growth and metastasis, while its role is unexplored in UC. To first elucidate the clinical significance of SULF1 transcript expression, real-time quantitative RT-PCR was performed in a pilot study of 24 UTUC and 24 UBUC fresh samples. We identified that increased SULF1 transcript abundance was associated with higher primary tumor (pT) status. By testing SULF1 immunoexpression in independent UTUC and UBUC cohorts consisted of 340 and 295 cases, respectively, high SULF1 expression was significantly associated with advanced pT and nodal status, higher histological grade and presence of vascular invasion in both UTUC and UBUC. In multivariate survival analyses, high SULF1 expression was independently associated with worse DSS (UTUC hazard ratio [HR] = 3.574, P < 0.001; UBUC HR = 2.523, P = 0.011) and MeFS (UTUC HR = 3.233, P < 0.001; UBUC HR = 1.851, P = 0.021). Furthermore, depletion of SULF1 expression by using RNA interference leaded to impaired cell proliferative, migratory, and invasive abilities in vitro. In addition, we further confirmed oncogenic role of SULF1 with gain-of function experiments. In conclusion, our findings implicate the oncogenic role of SULF1 expression in UC, suggesting SULF1 as a prognostic and therapeutic target of UC.


[Aspects of the use of medical devices for remote patient monitoring].
In the Russian Federation, it is legally determined that telemedicine technologies are considered as any actions with any data obtained during a remote communication between doctors (councils, consultations), remote medical patient monitoring, including using medical devices. Medical devices with a function of remote transmission of the measurement data are used to detect diseases in some workers that impede their duty performance. There are hardly any publications on the experience of using such devices in other aspects of medical care. The issue of medical devices enhanced with remote data transmission to ensure standard of measurements has been elaborated in great detail. In terms of applicability in telemedicine technologies, remote patient monitoring devices are used to measure blood pressure and human body temperature, detect alcohol vapors in the exhaled air, record electrocardiogram. There is also patent activity. In 2019-2023, remote transmission of the measurement data including by medical devices was protected by a large number of invention or utility model patents. Nowadays, there are no legal, technological barriers to the use of medical devices enhanced with remote data transmission in primary care. In 2023-2024, 8 constituent entities of the Russian Federation are conducting a pilot project on remote patient monitoring with medical devices.


Promiscuous Yet Specific: A Methionine-Aromatic Interaction Drives the Reaction Scope of the Family 1 Glycosyltransferase GmUGT88E3 from Soybean.
Family 1 glycosyltransferases (GT1s, UGTs) catalyze the regioselective glycosylation of natural products in a single step. We identified GmUGT88E3 as a particularly promising biocatalyst able to produce a variety of pure, single glycosidic products from polyphenols with high chemical yields. We investigated this particularly desirable duality toward specificity, i.e., promiscuous toward acceptors while regiospecific. Using high-field NMR, kinetic characterization, molecular dynamics simulations, and mutagenesis studies, we uncovered that the main molecular determinant of GmUGT88E3 specificity is a methionine-aromatic bridge, an interaction often present in protein structures but never reported for enzyme-substrate interactions. Here, mutating Met127 led to inactive proteins or 100-fold reduced activity.


Interpreting the implications of DNA ancestry tests.
Shopping for genetic information has become popular, but consumers may not understand what exactly they are buying. The retail DNA industry is forcing laypersons, academics, and medical and legal professionals alike to face the crossroads of genetics, law, and society. How will we decipher the meanings of the tests, determine the value of the information provided, or appropriately encourage or discourage various applications of that genetic information? When it comes to understanding the signs at the crossroads of disciplines, something is always potentially lost in translation. This article provides an overview of the retail DNA industry, addressing a few questions ripe for misinterpretation and confusion. It argues that the challenges posed by the retail DNA industry are both intelligible and manageable; optimally, multidisciplinary individuals would guide the way, steering the courts, legislature, laboratories, and clinics toward an adequate balance of consumer protection, autonomy, and understanding.