[A case of encephalomyeloneuritis and HTLV-I infection].
The patient, a 65-year-old woman, had liver cirrhosis, and had blood transfusion at the age of 49 and 56. Early in September, 1989, she gradually developed numbness of the legs, staggering gait, and apathy with hallucination. In October, she became incontinent and unable to stand, and was admitted to Konan Hospital. On admission, she was disoriented with poor comprehension. Cranial nerves were intact except for horizontal nystagmus on lateral gaze. She had generalized areflexia without pathological reflex. Muscular forces were fairly preserved. Superficial sensations were diminished in the upper limbs as well as below Th-7 level. Deep sensation was abolished in the distal parts of the extremities with athetotic finger postures on arm rising. She had urinary and fecal incontinence. Results of routine laboratory examinations were non-contributory. Chest CT scan and sputum cytology were normal. CSF contained one cell/microliter, 95 mg/dl of protein with positive oligoclonal IgG bands. Anti-HTLV-I antibody was positive in serum and CSF. Urodynamic studies showed neurogenic bladder of supranuclear type. MNCV was slightly decreased. SNAP and SEP were not evoked. On sural nerve biopsy, the density of myelinated fibers was 720/mm2, and that of unmyelinated fibers, 26,978/mm2. ABR and VEP were abnormal. EEG showed diffuse theta waves with paroxysmal delta and sharp waves. T2-weighted MR images of the brain showed patchy areas of high signal intensity in the cerebral white matter. Soon after administration of methylprednisolone, her consciousness became clear. EEG normalized in 4 months. Twenty months after the onset, she became ambulant with crutch, but still has dysuria and sensory deficits in the hands and lower limbs. The possible relationship between encephalomyeloneuritis and HTLV-I infection was discussed.


Reducing stillbirths in Ethiopia: Results of an intervention programme.
Previous studies from South Ethiopia have shown that interventions that focus on intrapartum care substantially reduce maternal mortality and there is a need to operationalize health packages that could reduce stillbirths. The aim of this paper is to evaluate if a programme that aimed to improve maternal health, and mainly focusing on strengthening intrapartum care, also would reduce the number of stillbirths, and to estimate if there are other indicators that explains high stillbirth rates. Our study used a "continuum of care" approach and focussed on providing essential antenatal and obstetric services in communities through health extension workers, at antenatal and health facility services. In this follow up study, which includes the same 38.312 births registered by community health workers, shows that interventions focusing on improved intrapartum care can also reduce stillbirths (by 46%; from 14.5 to 7.8 per 1000 births). Other risk factors for stillbirths are mainly related to complications during delivery and illnesses during pregnancy. We show that focusing on Comprehensive Emergency Obstetric Care and antenatal services reduces stillbirths. However, the study also underlines that illnesses during pregnancy and complications during delivery still represent the main risk factors for stillbirths. This indicates that obstetric care need still to be strengthened, should include the continuum of care from home to the health facility, make care accessible to all, and reduce delays.


The antihypertensive effect of ethyl acetate extract from red raspberry fruit in hypertensive rats.
To evaluate the antihypertensive effect of Xinjiang red raspberry fruit ethyl acetate extract (EER) on spontaneously hypertensive rats (SHR) and its possible mechanism from antioxidant perspective. The SHR rats were randomly divided into 3 groups, and treated with EER low dose (EERL, 100 mg/kg/d), high dose (EERH, 200 mg/kg/d), and water (SHR) through gastric gavage daily for 5 weeks. Another 8 age-matched male Wistar-Kyoto rats were used as normotensive group (WKY). The systolic blood pressure (SBP) was measured by noninvasive tail-cuff method once a week. At the end of the treatment, blood samples were collected and serum concentrations of nitric oxide (NO), superoxide dismutase (SOD), malondialchehyche (MDA), and plasma endothelin (ET) were determined. Treatment of SHR rats with EER lowered the blood pressure compared with that treated with water (SHR), and the high dose showed more significant reduction in blood pressure. Treatment of SHR rats with EER increased serum NO and SOD levels and lowered ET and MDA levels. As compared with control group, NO levels were increased significantly in EERL (P < 0.01), SOD was elevated more significantly in both EERL and EERH (P < 0.01); MDA was decreased significantly in EERH group (P < 0.05), whereas plasma ET decreased more significantly in the EERH group (P < 0.05). The red raspberry extracts demonstrated a dose-dependent antihypertensive effects in SHR and this may be related to increased NO activation and improved vascular endothelial dysfunction via antioxidation. These results confirmed that raspberries rich in polyphenols have potential cardiovascular protective effects.


A Pinus radiata AAA-ATPase, the expression of which increases with tree ageing.
Age-dependent changes in gene expression profiles were studied in vegetative Pinus radiata buds by means of differential display. Among several candidate cDNAs, a 327 bp fragment that shows high homology with an Arabidopsis thaliana 20S proteasome ATPase designated RPT5a was found. Northern hybridization confirmed that the accumulation of this transcript increases with tree ageing, suggesting a possible role of this AAA-ATPase gene in development-related specific proteolysis.


An exploratory study of the potential learning benefits for medical students in collaborative drawing: creativity, reflection and 'critical looking'.
Building on a series of higher educational arts/medicine initiatives, an interdisciplinary drawing module themed on the human body was developed for both year 3 Craft students and year 3 Medicine degree students. This became the subject of a research project exploring how the collaborative approach to drawing adopted on this module impacted on the students' learning. In this article, emphasis is given to issues thought to have most potential relevance to medical education. Using an ethnographic research design, the methods adopted were: direct observation of all aspects of the module sessions, audio and video recordings and photographs of the sessions, the incorporation of a semi-structured discussion at the end of each session, and anonymous student questionnaires. A number of key themes emerged. The complex, phased and multi-sensory nature of the 'critical looking' skills developed through the drawing exercises was seen as of potential value in medical education, being proposed as analogous to processes involved in clinical examination and diagnosis. The experience of interdisciplinary collaborative drawing was significant to the students as a creative, participatory and responsive form of learning. The emphasis on the physical experience of drawing and the thematic use of the human body as drawing subject led to reflective discussions about bodily knowledge and understanding. There were indications that students had a meta-cognitive awareness of the learning shifts that had occurred and the sessions provoked constructive self-reflective explorations of pre-professional identity. This preliminary study suggests, through the themes identified, that there may be potential learning outcomes for medical students in this model of interdisciplinary collaborative drawing of the human body. Further research is needed to explore their applicability and value to medical education. There is a need to explore in more depth the beliefs, motivations and learning styles of medical students opting for the module, the significance and weighting of different learning and teaching elements in the module and the impact of the learning on medical students in the immediate post-module phase.


Selective medium for the isolation and enumeration of Mycobacterium avium-intracellulare and M. scrofulaceum.
A medium for the selective isolation and enumeration of Mycobacterium avium-intracellulare and M. scrofulaceum (MAIS) was developed, based upon the ability of these mycobacteria to utilize Tween 80 as sole carbon source and grow optimally at pH 5.5 on a simple mineral salts medium. Representative MAIS strains had higher efficiencies of plating on the Tween 80 medium compared with Middlebrook 7H10. It was shown that nonmycobacterial organisms in natural waters had lower efficiencies of plating on the Tween 80 medium and smaller colonies, thus allowing direct isolation and enumeration of the slowly growing mycobacteria without overgrowth.


A genomic screen for angiosuppressor genes in the tumor endothelium identifies a multifaceted angiostatic role for bromodomain containing 7 (BRD7).
Tumor angiogenesis is characterized by deregulated gene expression in endothelial cells (EC). While studies until now have mainly focused on overexpressed genes in tumor endothelium, we here describe the identification of transcripts that are repressed in tumor endothelium and thus have potential suppressive effects on angiogenesis. We identified nineteen putative angiosuppressor genes, one of them being bromodomain containing 7 (BRD7), a gene that has been assigned tumor suppressor properties. BRD7 was studied in more detail, and we demonstrate that BRD7 expression is inversely related to EC activation. Ectopic expression of BRD7 resulted in a dramatic reduction of EC proliferation and viability. Furthermore, overexpression of BRD7 resulted in a bromodomain-dependent induction of NFκB-activity and NFκB-dependent gene expression, including ICAM1, enabling leukocyte-endothelial interactions. In silico functional annotation analysis of genome-wide expression data on BRD7 knockdown and overexpression revealed that the transcriptional signature of low BRD7 expressing cells is associated with increased angiogenesis (a.o. upregulation of angiopoietin-2, VEGF receptor-1 and neuropilin-1), cytokine activity (a.o. upregulation of CXCL1 and CXCL6), and a reduction of immune surveillance (TNF-α, NFκB, ICAM1). Thus, combining in silico and in vitro data reveals multiple pathways of angiosuppressor and anti-tumor activities of BRD7.


A contact-insensitive subpopulation in Syrian hamster cell cultures with a greater susceptibility to chemically induced neoplastic transformation.
We previously have identified a subpopulation of contact-insensitive (CS-) cells which lacks density-dependent inhibition of cell division in primary and low-passage cultures of Syrian hamster embryonic (SHE) fibroblastic cells. Further, we have shown that the proportion of these CS- cells declines as a result of the stable phenotypic conversion of the CS- cells to contact-sensitive (CS+) cells. To determine whether these transient CS- cells are more sensitive to carcinogenic/mutagenic perturbation, the susceptibility to neoplastic transformation and somatic mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was examined in clonally isolated cell cultures containing various proportions of CS- cells (0.02-4%). The frequencies of morphological transformation, focus formation, and neoplastic transformation showed a positive correlation to the proportion of CS- cells in the treated cultures. In contrast, the frequency of MNNG-induced somatic mutation at the Na+,K+-ATPase locus was similar among cultures varying in their proportion of CS- cells. Thus, there is a transient subpopulation of CS- cells in primary SHE cell cultures that is more susceptible to neoplastic transformation although equally susceptible to induced point mutation. This dissociation between somatic point mutation and neoplastic transformation indicates a fundamental difference in the nature of these two phenomena. A possible relationship between the propensity of CS- cells (versus CS+ cells) to carcinogen-induced neoplastic transformation and the state of differentiation of the CS- cells is discussed.


Association of low-density lipoprotein pattern with mortality after myocardial infarction: Insights from the TRIUMPH study.
Studies of incident coronary heart disease risk within low-density lipoprotein (LDL) subclass (small, dense vs large, buoyant) have shown mixed results. No prospective cohort study has examined the association of small, dense, or large, buoyant LDL with mortality after myocardial infarction (MI). The objective of the study was to examine association of LDL pattern after MI and death. In 2476 patients hospitalized for MI, LDL pattern (A [large, buoyant], A/B [mixed], and B [small, dense]) was established by ultracentrifugation using Vertical Auto Profile. Using time-to-event analysis, we examined the association with 5-year mortality within LDL patterns, after adjusting for important patient and treatment characteristics. We additionally adjusted for LDL cholesterol (LDL-C) and triglyceride levels and used directly measured LDL-C and non-high-density lipoprotein cholesterol as exposures. Patterns A, A/B, and B were present in 39%, 28%, and 33% of patients, respectively, with incident rates (per 1000 patient-years) of 50, 34, and 24 for all-cause and 24, 19, and 10 for CV mortality. The hazard ratios (95% confidence interval) with LDL patterns A/B and B compared with pattern A were 0.77 (0.61, 0.99) and 0.67 (0.51, 0.88) for all-cause, 0.94 (0.67, 1.33) and 0.69 (0.46, 1.03) for cardiovascular, and 0.64 (0.45, 0.91) and 0.65 (0.45, 0.93) for noncardiovascular mortalities, respectively. Results were similar when further adjusted for LDL-C and triglycerides, or with LDL-C and non-high-density lipoprotein cholesterol as exposures. Compared with LDL pattern A, pattern B was significantly associated with reduced all-cause and non-CV mortalities with a trend for lower CV mortality after MI, independent of LDL-C and triglycerides.


[Effectiveness of venlafaxine in the treatment of alcohol dependence with comorbid depression].
There are no conclusive data on the effectiveness of antidepressant drugs in the treatment of comorbid cases of alcohol dependence and depression. To determine the effectiveness of venlafaxine on depression and on severity (need of treatment) of alcohol dependence and related problems. Observational, open-label, multicenter, 24-week follow-up study. 90 outpatients with diagnosis of alcohol dependence and associated major depression disorder (DSMIV criteria). the Hamilton Rating Scale for Depression (HAM-D17), European Addiction Severity Index (EuropASI) and Clinical Global Impression, severity and improvement subscales, (CGI-S and CGI-I). Evaluations were performed at baseline and at weeks 2, 4, 8 and 24. Mean age 44.94+/-9.74 years; 73.3 % man. HAM-D17 mean scores significantly decreased from baseline (24.85+/-5.94) to week 24 (5.976+/-4.68) and at each of the follow-up visits vs previous visit (p < 0.0005). Significant decreases from baseline to week 24 were obtained in four areas of EuropASI: medical status (2.12+/-2.45 to 1.07+/-1.68), alcohol use (5.29+/-2.24 to 3.04+/-2.35), family/ social relationships (3.68+/-2.36 to 1.71+/-2.06) and psychiatric status (5.61+/-1.81 to 2.67+/-2.03). Tolerance was excellent or good in 76.7% of the patients. Venlafaxine demonstrated to be effective in the treatment of depressive alcoholic patients. Furthermore, it seems to be useful to decrease the severity of problems related with the alcohol use.


Characteristics of elderly persons seen in an urban psychiatric emergency room.
With the aging of the U.S. population, an increase in the use of the psychiatric emergency room by elderly persons is expected. In this study of 118 elderly persons seen in a general hospital psychiatric emergency room, half had a history of psychiatric illness. A third appeared to have chronic organic brain disease, and a third had affective disorders. Misprescription or misuse of medications was implicated in the problems of 27 patients. The problems of most patients were not emergencies; most had experienced symptoms of their presenting complaint for more than six months, finally overtaxing the coping capacities of their caregivers. The authors discuss factors that prevent elderly persons from seeking help on their own and emphasize the need for earlier interventions with elderly patients and for education of the public and professionals about issues, such as overmedication, that are important in the care of elderly persons.


Mild phenotype of knockouts of the major apurinic/apyrimidinic endonuclease APEX1 in a non-cancer human cell line.
The major human apurinic/apyrimidinic (AP) site endonuclease, APEX1, is a central player in the base excision DNA repair (BER) pathway and has a role in the regulation of DNA binding by transcription factors. In vertebrates, APEX1 knockouts are embryonic lethal, and only a handful of knockout cell lines are known. To facilitate studies of multiple functions of this protein in human cells, we have used the CRISPR/Cas9 system to knock out the APEX1 gene in a widely used non-cancer hypotriploid HEK 293FT cell line. Two stable knockout lines were obtained, one carrying two single-base deletion alleles and one single-base insertion allele in exon 3, another homozygous in the single-base insertion allele. Both mutations cause a frameshift that leads to premature translation termination before the start of the protein's catalytic domain. Both cell lines totally lacked the APEX1 protein and AP site-cleaving activity, and showed significantly lower levels of the APEX1 transcript. The APEX1-null cells were unable to support BER on uracil- or AP site-containing substrates. Phenotypically, they showed a moderately increased sensitivity to methyl methanesulfonate (MMS; ~2-fold lower EC50 compared with wild-type cells), and their background level of natural AP sites detected by the aldehyde-reactive probe was elevated ~1.5-2-fold. However, the knockout lines retained a nearly wild-type sensitivity to oxidizing agents hydrogen peroxide and potassium bromate. Interestingly, despite the increased MMS cytotoxicity, we observed no additional increase in AP sites in knockout cells upon MMS treatment, which could indicate their conversion into more toxic products in the absence of repair. Overall, the relatively mild cell phenotype in the absence of APEX1-dependent BER suggests that mammalian cells possess mechanisms of tolerance or alternative repair of AP sites. The knockout derivatives of the extensively characterized HEK 293FT cell line may provide a valuable tool for studies of APEX1 in DNA repair and beyond.


Fibronectin in the developing sea urchin embryo.
The presence of fibronectin in developing sea urchin embryos was studied uing immunofluorescence staining. The fluorescence pattern indicates that fibronectin is found on the cell surfaces and between cells in the blastula and gastrula stages, indicating that it plays a role in cell adhesion. Its presence on invaginating cells also suggests its involvement in morphogenesis during early development.


Effects of malnutrition and chronic reserpine treatment on pancreatic exocrine function.
The chronically reserpine-treated rat, an experimental model for cystic fibrosis, exhibits generalized exocrinopathy, impaired pancreatic secretion, and decreased pancreatic amylase. Although chronic reserpine treatment induces malnutrition by decreasing food consumption and growth, the effects of this malnutrition per se on the exocrine pancreas have not been considered. In this study, the effects of chronic reserpine treatment and malnutrition on the exocrine pancreas were determined using pair-fed controls. Male, Sprague-Dawley rats were treated daily subcutaneously for 5 to 7 days with: no injection (control), 1.0 ml/kg vehicle or sham (control-sham, pair fed-sham), or 0.5 mg/kg reserpine (chronically reserpine-treated). Both chronic reserpine-treatment and pair-feeding significantly decreased food consumption (40%), body weight (51 and 59%), total pancreatic amylase (49 and 56%) and specific amylase activity (62 and 61%), pancreatic protein (65 and 75%), and pancreatic weights (62 and 65%) compared to controls. These decreases, however, were comparable between the chronically reserpine-treated and pair fed-sham rats. In contrast, the secretory response to the biologically active cholecystokinin analog cholecystokinin octapeptide was significantly attenuated in isolated pancreatic acini prepared from reserpine-treated rats compared to that from either control or pair-fed sham rats. Malnutrition decreased pancreatic amylase activity and protein comparably to reserpine treatment, but only partially attenuated the secretory response to cholecystokinin octapeptide. Based on the results of this study, pair-fed controls should be used to distinguish between the effects of reserpine alone and the induced malnutrition on pancreatic exocrine function in studies of this experimental model of cystic fibrosis.


Assessment of embryo morphology and developmental dynamics by time-lapse microscopy: is there a relation to implantation and ploidy?
Time-lapse microscopy (TLM) is an exciting novel technology with great potential for enhancing embryo selection in the embryology laboratory. This non-invasive objective assessment of embryos has provided a new tool for predicting embryo development and implantation potential. TLM detects several morphological phenomena that are often missed with static observations using conventional incubators, such as irregular divisions, blastocyst collapse and re-expansion, timing of blastocoel appearance, and timing of formation and internalization of fragments. Nevertheless, it should be recognized that conventional morphological assessment has been widely accepted as the gold standard by most embryologists. TLM can enhance conventional morphological assessments to improve embryo selection and subsequent reproductive outcomes. Furthermore, morphokinetic parameters can aid in differentiating between euploid and aneuploid embryos, although they are not sufficiently accurate to replace preimplantation genetic testing for aneuploidy. Morphokinetic assessment together with chromosomal screening may ultimately help identify euploid embryos with the highest developmental potential.


Can Serum Nitrosoproteome Predict Longevity of Aged Women?
Aging is characterized by increase in reactive oxygen (ROS) and nitrogen (RNS) species, key factors of cardiac failure and disuse-induced muscle atrophy. This study focused on serum nitroproteome as a trait of longevity by adopting two complementary gel-based techniques: two-dimensional differential in gel electrophoresis (2-D DIGE) and Nitro-DIGE coupled with mass spectrometry of albumin-depleted serum of aged (A, n = 15) and centenarian (C, n = 15) versus young females (Y, n = 15). Results indicate spots differently expressed in A and C compared to Y and spots changed in A vs. C. Nitro-DIGE revealed nitrosated protein spots in A and C compared to Y and spots changed in A vs. C only (p-value < 0.01). Nitro-proteoforms of alpha-1-antitripsin (SERPINA1), alpha-1-antichimotripsin (SERPINA3), ceruloplasmin (CP), 13 proteoforms of haptoglobin (HP), and inactive glycosyltransferase 25 family member 3 (CERCAM) increased in A vs. Y and C. Conversely, nitrosation levels decreased in C vs. Y and A, for immunoglobulin light chain 1 (IGLC1), serotransferrin (TF), transthyretin (TTR), and vitamin D-binding protein (VDBP). Immunoblottings of alcohol dehydrogenase 5/S-nitrosoglutathione reductase (ADH5/GSNOR) and thioredoxin reductase 1 (TRXR1) indicated lower levels of ADH5 in A vs. Y and C, whereas TRXR1 decreased in A and C in comparison to Y. In conclusion, the study identified putative markers in C of healthy aging and high levels of ADH5/GSNOR that can sustain the denitrosylase activity, promoting longevity.


Anterior cruciate ligament injuries in the skeletally immature patient.
Injuries to the anterior cruciate ligament in children and youth pose a complex clinical challenge. The literature does not provide the clinician with a well-defined natural history or pathoanatomy, nor does it show agreement as to the outcomes of various treatment choices. Decision making is made difficult by the patients' skeletal and emotional immaturity and by associated intra-articular fracture, physeal fracture, and combination injuries. In an effort to place the problem of ACL injury in children and youth into some perspective and to present an algorithm for management, this article reviews issues in its diagnosis, classification, anatomy, mechanism of injury, and management.


Electron-microscopic and immunochemical analysis of kinetochore microtubules after ultraviolet microbeam irradiation of kinetochores.
We used an ultraviolet microbeam to irradiate kinetochores of chromosomes in crane-fly spermatocytes. We used one of two doses, low (0.106 erg microns-2) or high (0.301 erg microns-2), and then studied the microtubules in those spindles using electron microscopy or immunofluorescence microscopy. After irradiation with low doses microtubules are present as usual, with normal fluorescence and in normal numbers. After irradiation with high doses microtubules are no longer associated with the irradiated kinetochore. After irradiation with either dose, non-kinetochore microtubules are in smaller numbers in the irradiated half-spindle than in the non-irradiated half-spindle or in non-irradiated cells. Since irradiation with low doses alters interchromosomal 'signals', but microtubules remain attached to the kinetochore, we argue that low doses of ultraviolet light damage a signal-related function of kinetochores without altering the ability of the kinetochores to bind microtubules.


A single mutation alters production and discrimination of Drosophila sex pheromones.
The evolution of communication is a fundamental biological problem. The genetic control of the signal and its reception must be tightly coadapted, especially in inter-individual sexual communication. However, there is very little experimental evidence for tight genetic linkage connecting the emission of a signal and its reception. We found that a single genomic transposon inserted in the desatl gene of Drosophila melanogaster simultaneously affected the emission and the perception of sex-specific signals. This mutation greatly decreased the production of unsaturated hydrocarbons on the cuticle of mature flies of both sexes. These substances represent the sex pheromones necessary for mate discrimination: control males could not discriminate the sex of mutant desatl flies. Moreover, mutant males were unable to discriminate the sex pheromones of con-trol flies. Expression of desatl was found in the peripheral tissues that produce and detect sex pheromones. Excision of the transposon rescued both the production and discrimination phenotypes, but the two effects did not always coincide. This indicates that the emission and perception of pheromones are coded by differ-ent products of the same gene, reflecting the pleiotropic activity of desatl .


Salinomycin and IR780-loaded upconversion nanoparticles influence biological behavior of liver cancer stem cells by persistently activating the MAPK signaling pathway.
The combination of chemotherapy and phototherapy has emerged as a promising therapeutic approach for enhancing the efficacy of cancer treatment and mitigating drug resistance. Salinomycin (SAL), a polyether antibiotic, exhibits potent cytotoxicity against chemotherapy-resistant cancer cells. IR780 iodide, a novel photosensitive reagent with excellent near-infrared (NIR) light absorption and photothermal conversion abilities, is suitable for use in photothermal therapy for cancers. However, both SAL and IR780 exhibit hydrophobic properties that limit their clinical applicability. Upconversion nanoparticles (UCNPs) are an emerging class of fluorescent probe materials capable of emitting high-energy photons upon excitation by low-energy NIR light. The UCNPs not only function as nanocarriers for drug delivery but also serve as light transducers to activate photosensitizers for deep-tissue photodynamic therapy. Here, to enhance the targeting and bioavailability of hydrophobic drugs in liver cancer stem cells (LCSCs), we employ distearoyl phosphorethanolamine-polyethylene glycol (DSPE-PEG) to encapsulate SAL and IR780 on the surface of UCNPs. Cell viability was evaluated using the CCK-8 assay. Cell migration was assessed by the Transwell Boyden Chamber. The activation of the mitogen-activated protein kinase (MAPK) signaling pathway was measured via western blot. The results demonstrated successful loading of both IR780 and SAL onto the UCNPs, and the SAL and IR780-loaded UCNPs (UISP) exhibited a robust photothermal effect under NIR light irradiation. The UISP effectively inhibited the viability of HCCLM3 and LCSCs. Under NIR light irradiation, the UISP further suppressed HCCLM3 viability but had no impact on LCSC viability; however, it could further inhibit LCSC migration. Meanwhile, under NIR light irradiation, the UISP persistently activated the MAPK pathway more significantly in LCSCs. These findings suggest that exposure to NIR light results in persistent activation of the MAPK pathway by UISP, thereby influencing the biological behavior of LCSCs and enhancing their therapeutic efficacy against liver cancer.


Alcohol and absorption from the small intestine. 1. Impairment of absorption from the small intestine in alcoholics.
An absorption screen was performed in 10 chronic alcoholic patients within a few days of admission due to an acute alcoholic episode. Impaired absorption of d-Xylose was noted in three patients and low leucocyte ascorbic acid and serum folate levels in five. No abnormality was detected in jejunal histology. The absorption of water and electrolytes from the jejunum was studied in these patients using a triple-lumen tube perfusion system. The mean rate of absorption of water in the alcoholic subjects (50-0 +/- 2-3 ml/h) was significantly lower (P less than 0-001) than the mean value in 14 healthy control subjects (205 +/- 15-9 ml/h). A significant reduction of Na+ and Cl-absorption was also demonstrated in the alcoholic subjects. These results indicate that patients with acute-on-chronic alcoholism may have a function impairment of water and electrolyte absorption from the jejunum. This may, in part, account for some of the nutritional deficiencies in such patients and for symptoms such as diarrhoea which may be present.


Oral lichen planus and the p53 family: what do we know?
Oral lichen planus (OLP) is a relatively common chronic disease of the oral mucosa for which the aetiopathogenesis is not fully understood. It mainly affects middle aged and elderly. The finding of autoantibodies against p63, a member of the p53 family, is a strong indication of autoimmunity as a causative or contributing factor. The WHO classified OLP as a potentially malignant disorder, but still there is an ongoing debate in the literature on this subject. The TP53 gene encodes a tumour suppressor protein that is involved in induction of cell-cycle arrest or apoptosis of DNA-damaged cells. The p63 gene encodes six different proteins that are crucial for formation of the oral mucosa and skin. The coordinated stabilization of p53 and decreased expression of p63 seen in OLP cause induction of apoptosis enabling removal of DNA-damaged cells. In view of the complexity of cancerogenesis, no firm statement can at present be made about the relevance of the observed relationship between p53 and p63 and the possible malignant transformation of OLP.


Exploring the time window for causal inference and the multisensory integration of actions and their visual effects.
Successful computer use requires the operator to link the movement of the cursor to that of his or her hand. Previous studies suggest that the brain establishes this perceptual link through multisensory integration, whereby the causality evidence that drives the integration is provided by the correlated hand and cursor movement trajectories. Here, we explored the temporal window during which this causality evidence is effective. We used a basic cursor-control task, in which participants performed out-and-back reaching movements with their hand on a digitizer tablet. A corresponding cursor movement could be shown on a monitor, yet slightly rotated by an angle that varied from trial to trial. Upon completion of the backward movement, participants judged the endpoint of the outward hand or cursor movement. The mutually biased judgements that typically result reflect the integration of the proprioceptive information on hand endpoint with the visual information on cursor endpoint. We here manipulated the time period during which the cursor was visible, thereby selectively providing causality evidence either before or after sensory information regarding the to-be-judged movement endpoint was available. Specifically, the cursor was visible either during the outward or backward hand movement (conditions Out and Back, respectively). Our data revealed reduced integration in the condition Back compared with the condition Out, suggesting that causality evidence available before the to-be-judged movement endpoint is more powerful than later evidence in determining how strongly the brain integrates the endpoint information. This finding further suggests that sensory integration is not delayed until a judgement is requested.


Do squid breathe through their skin?
Squid are thought to obtain a large portion of their oxygen via simple diffusion across the skin in addition to uptake at the gills. Although this hypothesis has support from indirect evidence and is widely accepted, no empirical examinations have been conducted to assess the validity of this hypothesis. In this study, we examined cutaneous respiration in two squid species, Doryteuthis pealeii and Lolliguncula brevis, using a divided chamber to physically separate the mantle cavity and gills from the outer mantle surface. We measured oxygen consumption and ammonia excretion rates in the two compartments and found that, at rest, squid only obtain enough oxygen cutaneously to meet the demand of the skin tissue locally (12% of total) and excrete little ammonia across the skin. The majority of oxygen is obtained via the traditional branchial pathway. In light of these findings, we re-examine and discuss the indirect evidence that has supported the cutaneous respiration hypothesis.


Revealing the metabolic profile of brain tumors for diagnosis purposes.
The metabolic behavior of complex brain tumors, like Gliomas and Meningiomas, with respect to their type and grade was investigated in this paper. Towards this direction the smallest set of the most representative metabolic markers for each brain tumor type was identified, using ratios of peak areas of well established metabolites, from (1)H-MRSI (Proton Magnetic Resonance Spectroscopy Imaging) data of 24 patients and 4 healthy volunteers. A feature selection method that embeds Fisher's filter criterion into a wrapper selection scheme was applied; Support Vector Machine (SVM) and Least Squares-SVM (LS-SVM) classifiers were used to evaluate the ratio markers classification significance. The area under the Receiver Operating Characteristic curve (AUROC) was adopted to evaluate the classification significance. It is found that the NAA/CHO, CHO/S, MI/S ratios can be used to discriminate Gliomas and Meningiomas from Healthy tissue with AUROC greater than 0.98. Ratios CHO/S, CRE/S, MI/S, LAC/CRE, ALA/CRE, ALA/S and LIPS/CRE can identify type and grade differences in Gliomas giving AUROC greater than 0.98 apart from the scheme of Gliomas grade II vs grade III where 0.84 was recorded due to high heterogeneity. Finally NAA/CRE, NAA/S, CHO/S, MI/S and ALA/S manage to discriminate Gliomas from Meningiomas providing AUROC exceeding 0.90.


Presynaptic inhibition preferentially reduces in NMDA receptor-mediated component of transmission in rat midbrain dopamine neurons.
We used patch pipettes to record whole-cell currents from single dopamine neurons in slices of rat midbrain. Pharmacological methods were used to isolate EPSCs evoked by focal electrical stimulation. Baclofen was significantly more potent for inhibiting NMDA receptor-mediated EPSCs (IC50=0.24 microM) compared with inhibition of EPSCs mediated by AMPA receptors (IC50=1.72 microM). The increased potency of baclofen for inhibiting the NMDA component persisted in superfusate that contained zero Mg2+ and when postsynaptic K+ conductances were reduced by Cs+ and QX-314. Effects of baclofen on EPSCs were blocked by the GABA(B) receptor antagonist CGP-35348. Adenosine was 20 fold more potent for reducing the NMDA component of transmission (IC50=31 microM) compared with inhibition of AMPA receptor-mediated EPSCs (IC50=654 microM). Effects of adenosine on EPSCs were blocked by the A1 receptor antagonist DPCPX. Both baclofen and adenosine significantly increased the ratio of EPSCs in paired-pulse studies, suggesting presynaptic sites of action. Although adenosine (1 mM) did not reduce currents evoked by exogenous NMDA (10 microM), baclofen (1 microM) reduced NMDA currents by 29%. Neither baclofen nor adenosine altered currents evoked by exogenous AMPA (1 microM). We conclude that adenosine acts at presynaptic A1 receptors to cause a preferential reduction in the NMDA component of synaptic transmission. In contrast, baclofen preferentially reduces NMDA EPSCs by acting at both pre- and postsynaptic GABA(B) receptors. By regulating NMDA receptor function, A1 and GABA(B) receptors may play important roles in regulating the excitability of dopamine neurons.


A timeline of bacterial and archaeal diversification in the ocean.
Microbial plankton play a central role in marine biogeochemical cycles, but the timing in which abundant lineages diversified into ocean environments remains unclear. Here, we reconstructed the timeline in which major clades of bacteria and archaea colonized the ocean using a high-resolution benchmarked phylogenetic tree that allows for simultaneous and direct comparison of the ages of multiple divergent lineages. Our findings show that the diversification of the most prevalent marine clades spans throughout a period of 2.2 Ga, with most clades colonizing the ocean during the last 800 million years. The oldest clades - SAR202, SAR324, Ca. Marinimicrobia, and Marine Group II - diversified around the time of the Great Oxidation Event, during which oxygen concentration increased but remained at microaerophilic levels throughout the Mid-Proterozoic, consistent with the prevalence of some clades within these groups in oxygen minimum zones today. We found the diversification of the prevalent heterotrophic marine clades SAR11, SAR116, SAR92, SAR86, and Roseobacter as well as the Marine Group I to occur near to the Neoproterozoic Oxygenation Event (0.8-0.4 Ga). The diversification of these clades is concomitant with an overall increase of oxygen and nutrients in the ocean at this time, as well as the diversification of eukaryotic algae, consistent with the previous hypothesis that the diversification of heterotrophic bacteria is linked to the emergence of large eukaryotic phytoplankton. The youngest clades correspond to the widespread phototrophic clades Prochlorococcus, Synechococcus, and Crocosphaera, whose diversification happened after the Phanerozoic Oxidation Event (0.45-0.4 Ga), in which oxygen concentrations had already reached their modern levels in the atmosphere and the ocean. Our work clarifies the timing at which abundant lineages of bacteria and archaea colonized the ocean, thereby providing key insights into the evolutionary history of lineages that comprise the majority of prokaryotic biomass in the modern ocean.


Emulating synaptic behavior in surface-functionalized MoS2 through modulation of interfacial charge transfer via external stimuli.
Neuromorphic computing requires materials able to yield electronic switching behavior in response to external stimuli. Transition-metal dichalcogenides surfaces covered by partial or full monolayers of molecular species have shown promise due to their potential for tunable interfacial charge transfer. Here, we demonstrate a class of molecules able to position MoS2 surfaces on the cusp of electronic instabilities. Density functional theory (DFT) calculations and ab initio molecular dynamics simulations are used to study the interaction of four reduced pyridinium-derived pi-conjugated molecules with the pristine basal planes of MoS2, by exploring the dynamical evolution of the system at room temperature with regards to the effective band gap, radius of gyration (rog), and charge transfer. Computed rog profiles show that low concentrations of small reduced methyl viologen molecules have high mobilities on top of the surface of the basal plane at room temperature leading to unstable surface deposition, whereas a full monolayer of larger fused-ring molecules deposited on the basal surface shows greater thermal stability. DFT analyses show these larger reduced pyridinium derivatives promote n-type doping on the basal plane due to a built-in electric field, which can be systematically tuned to induce a switching effect, opening and closing a bandgap and providing a fundamental means of driving electronic instabilities needed for emulating neuronal functionality.


Hox homeodomain proteins exhibit selective complex stabilities with Pbx and DNA.
Eight of the nine homeobox genes of the Hoxb locus encode proteins which contain a conserved hexapeptide motif upstream from the homeodomain. All eight proteins (Hoxb-1-Hoxb-8) bind to a target oligonucleotide in the presence of Pbx1a under conditions where minimal or no binding is detected for the Hox or Pbx1a proteins alone. The stabilities of the Hox-Pbx1a-DNA complexes vary >100-fold, with the proteins from the middle of the locus (Hoxb-5 and Hoxb-6) forming very stable complexes, while Hoxb-4, Hoxb-7 and Hoxb-8 form complexes of intermediate stability and proteins at the 3'-side of the locus (Hoxb-1-Hoxb-3) form complexes which are very unstable. Although Hox-b proteins containing longer linker sequences between the hexapeptide and homeodomains formed unstable complexes, shortening the linker did not confer complex stability. Homeodomain swapping experiments revealed that this motif does not independently determine complex stability. Naturally occurring variations within the hexapeptides of specific Hox proteins also do not explain complex stability differences. However, two core amino acids (tryptophan and methionine) which are absolutely conserved within the hexapeptide domains appear to be required for complex formation. Removal of N- and C-terminal flanking regions did not influence complex stability and the members of paralog group 4 (Hoxa-4, b-4, c-4 and d-4), which share highly conserved hexapeptides, linkers and homeodomains but different flanking regions, form complexes of similar stability. These data suggest that the structural features of Hox proteins which determine Hox-Pbx1a-DNA complex stability reside within the precise structural relationships between the homeodomain, hexapeptide and linker regions.


Lipopolysaccharide-binding protein and soluble CD14 in the vitreous fluid of patients with proliferative diabetic retinopathy.
The purpose of this study was to compare intravitreous levels of lipopolysaccharide-binding protein and soluble CD14 (sCD14) between patients with proliferative diabetic retinopathy (PDR) and nondiabetic subjects. This study included 19 consecutive Type 2 diabetic patients with PDR in whom a vitrectomy was performed. Sixteen vitreous humors from nondiabetic patients matched by age, with idiopathic macular holes, were selected from our vitreous bank and used as a control group. Lipopolysaccharide-binding protein was assessed by enzyme-linked immunosorbent assay and sCD14 by a solid-phase enzyme-amplified sensitive immunoassay. Lipopolysaccharide-binding protein and sCD14 levels were significantly higher in patients with PDR than in the control group (lipopolysaccharide-binding protein, P < 0.001; sCD14, P < 0.01). After correcting for vitreal proteins, the results remained significantly higher in patients with PDR. No differences in serum levels were observed, and we did not find any correlation between serum and vitreous levels. A direct correlation between lipopolysaccharide-binding protein and sCD14 was detected in the vitreous fluid (r = 0.57; P < 0.001) but not in the plasma. Finally, a significant correlation between intravitreal levels of both lipopolysaccharide-binding protein and sCD14 and interleukin-8 and monocyte chemotactic protein-1 was also detected. Lipopolysaccharide-binding protein and sCD14 are elevated in the vitreous fluid of patients with PDR and thus may play a role in the innate immune response triggered by the inflammatory injury characteristic of PDR.


Effects of reference pricing in pharmaceutical markets: a review.
This work aims to provide a systematic and updated survey of original scientific studies on the effect of the introduction of reference pricing (RP) policies in Organisation for Economic Co-operation and Development (OECD) countries. We searched PubMed, EconLit and Web of Knowledge for articles on RP. We reviewed studies that met the inclusion criteria established in the search strategy. From a total of 468 references, we selected the 35 that met all of the inclusion criteria. Some common themes emerged in the literature. The first was that RP was generally associated with a decrease in the prices of the drugs subject to the policy. In particular, price drops seem to have been experienced in virtually every country that implemented a generic RP (GRP) policy. A GRP policy applies only to products with expired patents and generic competition, and clusters drugs according to chemical equivalence (same form and active compound). More significant price decreases were observed in the sub-markets in which drugs were already facing generic competition prior to RP. Price drops varied widely according to the amount of generic competition and industrial strategies: brand-named drugs originally priced above RP values decreased their prices to a greater extent. A second common theme was that both therapeutic RP (TRP) and GRP have been associated with significant and consistent savings in the first years of application. A third general result is that generic market shares significantly increased whenever the firms producing brand-named drugs did not adopt one of the following strategies: lowering prices to RP values; launching new dosages and/or formulations; or marketing substitute drugs still under patent protection. Finally, concerning TRP, although more evidence is needed, studies based on a large number of patient-level observations showed no association between the RP policy and health outcomes.


[Comparative analysis of the major protective antigens production in Vibrio cholerae recombinant and producer strains of the classical biovar].
The comparative study of 4 constructed protective antigen producing strains of the classical biovar and V. cholerae strains 569 B Inaba and M41 Ogawa, used in manufacturing the cholera chemical vaccine "cholerogen-toxoid", was carried out. The study revealed that V. cholerae plasmid strains 2414 Ogawa, 2415 Inaba and nonplasmid strains 2416 Ogawa, 2417 Inaba had a higher level of production of the main protective antrigens in comparison with producer strains. They also synthesized much more (4-5 fold) cholera toxin, toxin co-regulated adhesion pili, contained protein OmpU in their outer membrane, exceeded 2- to 3-fold in the synthesis of pathogenicity enzymes (proteases, phospholipases) and synthesized the same amounts of 01 antigen, serovars Inaba and Ogawa. The use of the newly created protective-antigen producing strains in vaccine manufacturing could facilitate the preparation of a more effective cholera chemical vaccine "cholerogen-toxoid".


Lysosomal and free acid phosphatase in salivary glands of chironomus tentans.
In cells of salivary glands of last-instar larvae of Chironomus tentans, acid phosphatase activity is bound to (probable) lysosomes and a few other cell organelles. At the end of the pupal molt the salivary gland breaks down. While acid phosphatase in areas of nondegenerated cells is still restricted to the structures mentioned, in degenerated areas the enzyme is freely distributed in the cytoplasm.


Diabetic retinopathy care--an international quality comparison.
The purpose of this paper is to study the quality indicator appropriateness and use it for international quality comparison on diabetic retinopathy (DR) patient care process in one American and one Dutch eye hospital. A 17-item DR quality indicator set was composed based on a literature review and systematically applied in two hospitals. Qualitative analysis entailed document study and 12 semi-structured face-to-face interviews with ophthalmologists, managers, and board members of the two hospitals. While the medical-clinical approach to DR treatment in both hospitals was similar, differences were found in quality of care perception and operationalization. Neither hospital systematically used outcome indicators for DR care. On the process level, the authors found larger differences. Similarities and differences were found in the structure of both hospitals. The hospitals' particular contexts influenced the interpretation and use of quality indicators. Although quality indicators and quality comparison between hospitals are increasingly used in international settings, important local differences influence their application. Context should be taken into account. Since that context is locally bound and directly linked to hospital setting, caution should be used interpreting the results of quality comparison studies. International quality comparison is increasingly suggested as a useful way to improve healthcare. Little is known, however, about the appropriateness and use of quality indicators in local hospital care practices.


Application of ischaemia-free liver transplantation improves prognosis of patients with steatotic donor livers - a retrospective study.
The use of steatotic livers in liver transplantation (LT) is controversial. Ischaemia-free liver transplantation (IFLT) has obvious advantages for the recovery of allograft function. The aim of this study was to examine the effect of liver grafts with steatosis on outcome and the effect of IFLT with steatotic livers. 360 patients with LT were enrolled in this study. Perioperative characteristics and differences in outcome among different grades of steatotic groups, and between the IFLT and conventional LT (CLT) groups were analysed. Occurrence of early allograft dysfunction (EAD; 50%) and primary nonfunction (PNF; 20%) was significantly higher in the severe steatosis group (P < 0.001 and <0.001, respectively). Survival rate is significantly low in severe steatosis group (3-year: 60%, P = 0.0039). The IFLT group had a significantly lower occurrence of EAD than the CLT group (0% vs. 60%, P = 0.01). The level of postoperative peak AST, GGT and creatine were significantly lower in IFLT group (P = 0.009, 0.032 and 0.024, respectively). In multivariable analysis, IFLT and EAD were independent factors affecting postoperative survival. Severe steatotic livers lead to severe complications and poor outcomes in LT. IFLT has obvious advantages for reducing the rate of EAD in LT with steatotic livers.


Summer day-roost selection by eastern red bats varies between areas with different land-use histories.
The eastern red bat (Lasiurus borealis) is widely considered to be in decline, inspiring interest in identifying important habitats for conservation in the eastern United States. Unfortunately, knowledge of important day-roosting habitats is lacking for much of the species' range. We examined patterns of day-roost selection by male and female eastern red bats at two study sites in southeastern Ohio, U. S. A, to help fill this information gap. We radio-tagged 28 male and 25 female bats during the summers of 2016-2019 and located 53 male and 74 female roosts. Day-roost selection differed between sexes and study areas. In a mostly even-aged forest with significant historical disturbance, we found males and females roosting in trees located at higher elevations, with no clear selection based on tree or stand characteristics. Specifically, males selected trees with larger diameters located at lower, cooler elevations than females, which selected smaller diameter trees found at higher, warmer elevations. However, in a forest with less historical disturbance and more structural diversity, we found sexes differed in how they selected from available habitats. These data show that heterogeneity in environmental conditions can lead to different patterns in selection, even between sites located within a small geographic area. They also show that eastern red bats sexually segregate on the local landscape in the presence of diverse forest conditions but may not do so in the absence of such diversity. We recommend managing forests to maintain structural diversity across an elevational gradient to provide male and female eastern red bats with suitable day-roosting habitat in southeast Ohio.


[Immunologic status of alcoholic patients with pulmonary tuberculosis].
Monoclonal antibodies were used to study the T and B immunity systems in 60 alcoholic patients suffering from pulmonary tuberculosis. The patients with that combined pathology demonstrated marked immunodeficiency as regards the T system and an increase of humoral immunity. It is assumed that one of the main causes of immunodeficiency lies in the blockade of the differentiation of T cells at the stage of T prothymocytes. The rise of the level of humoral immunity is connected to a decline of T suppressors. Administration of thymalin to patients with associated alcoholism and pulmonary tuberculosis was found to be highly effective.


Interleukin-30 subverts prostate cancer-endothelium crosstalk by fostering angiogenesis and activating immunoregulatory and oncogenic signaling pathways.
Cancer-endothelial interplay is crucial for tumor behavior, yet the molecular mechanisms involved are largely unknown. Interleukin(IL)-30, which is expressed as a membrane-anchored cytokine by human prostate cancer (PC) cells, promotes PC vascularization and progression, but the underlying mechanisms have yet to be fully explored. PC-endothelial cell (EC) interactions were investigated, after coculture, by flow cytometry, transcriptional profiling, western blot, and ELISA assays. Proteome profiler phospho-kinase array unveiled the molecular pathways involved. The role of tumor-derived IL30 on the endothelium's capacity to generate autocrine circuits and vascular budding was determined following IL30 overexpression, by gene transfection, or its deletion by CRISPR/Cas9 genome editing. Clinical value of the experimental findings was determined through immunopathological study of experimental and patient-derived PC samples, and bioinformatics of gene expression profiles from PC patients. Contact with PC cells favors EC proliferation and production of angiogenic and angiocrine factors, which are boosted by PC expression of IL30, that feeds autocrine loops, mediated by IGF1, EDN1, ANG and CXCL10, and promotes vascular budding and inflammation, via phosphorylation of multiple signaling proteins, such as Src, Yes, STAT3, STAT6, RSK1/2, c-Jun, AKT and, primarily CREB, GSK-3α/β, HSP60 and p53. Deletion of the IL30 gene in PC cells inhibits endothelial expression of IGF1, EDN1, ANG and CXCL10 and substantially impairs tumor angiogenesis. In its interaction with IL30-overexpressing PC cells the endothelium boosts their expression of a wide range of immunity regulatory genes, including CCL28, CCL4, CCL5, CCR2, CCR7, CXCR4, IL10, IL13, IL17A, FASLG, IDO1, KITLG, TNFA, TNFSF10 and PDCD1, and cancer driver genes, including BCL2, CCND2, EGR3, IL6, VEGFA, KLK3, PTGS1, LGALS4, GNRH1 and SHBG. Immunopathological analyses of PC xenografts and in silico investigation of 1116 PC cases, from the Prostate Cancer Transcriptome Atlas, confirmed the correlation between the expression of IL30 and that of both pro-inflammatory genes, NOS2, TNFA, CXCR5 and IL12B, and cancer driver genes, LGALS4, GNRH1 and SHBG, which was validated in a cohort of 80 PC patients. IL30 regulates the crosstalk between PC and EC and reshapes their transcriptional profiles, triggering angiogenic, immunoregulatory and oncogenic gene expression programs. These findings highlight the angiostatic and oncostatic efficacy of targeting IL30 to fight PC.


Pharmacological approach to increasing the retention of radiation-induced γ-H2AX foci using phosphatase inhibitors: significance in radiation biodosimetry.
In a scenario of accidental mass radiation exposure transportation and analysis of samples may take some time, resulting in loss of biomarker information over this period. The present study aims to use phosphatase inhibitors for longer retention of focal signals to adopt γ-H2AX as a biodosimetric biomarker for the management of early triage. Peripheral blood lymphocytes isolated from healthy individuals were irradiated in vitro with x-rays and γ-H2AX foci were analysed using fluorescent microscopy and flow cytometric methods. Further, the effect of protein phosphatase 2A inhibitors such as calyculin A, fostriecin and okadiac acid on the retention of foci was studied. Fluorescent microscopy was found to be a more sensitive method than flow cytometry. Calyculin A showed significant retention of focal signals at 6 h with 1.5-fold increased retention compared to radiation alone; this may prove beneficial in early triage management because of a better dose approximation.


Blueberry floral probiotics: Nectar microbes inhibit the growth of Colletotrichum pathogens.
To identify whether microorganisms isolated from blueberry flowers can inhibit the growth of Colletotrichum, an opportunistic plant pathogen that infects flowers and threatens yields, and to assess the impacts of floral microbes and Colletotrichum pathogens on artificial nectar sugars and honey bee consumption. The growth inhibition of Colletotrichum (C. acutatum, C. fioriniae, C. gloeosporioides) was screened using both artificial nectar co-culture and dual culture plate assays. All candidate nectar microbes were screened for antagonism against a single C. acutatum isolate. Then, the top four candidate nectar microbes showing strongest inhibition of C. acutatum (Neokomagataea thailandica, N. tanensis, Metschnikowia rancensis, and Symmetrospora symmetrica) were evaluated for antagonism against three additional C. acutatum isolates, and single isolates of both C. fioriniae and C. gloeosporioides. In artificial nectar assays, single and three-species cultures inhibited growth of two of four C. acutatum isolates by ca. 60%, but growth of other Colletotrichum species was not affected. In dual culture plate assays, inhibition was observed for all Colletotrichum species for at least three of four selected microbial antagonists (13‒53%). Neither honey bee consumption of nectar nor nectar sugar concentrations were affected by any microbe or pathogen tested. Selected floral microbes inhibited growth of all Colletotrichum species in vitro, although the degree of inhibition was specific to the assay and pathogen examined. In all microbial treatments, nectar sugars were preserved, and honey bee preference was not affected.


A Double Hit to Ubiquitination Leading to a New Diagnosis of VEXAS Syndrome.
VEXAS (vacuoles, E1 enzyme, X-linked, auto-inflammatory, somatic) syndrome is a newly defined illness that bridges hematology, oncology, and rheumatology. Its pathophysiology originates in a mutation in the UBA1 gene that leads to a defect in ubiquitination resulting in a severe systemic inflammatory syndrome. It is associated with significant morbidity and mortality; however, data are scarce due to limited cases described in the literature. Here we describe a case of a male in his 60s who was referred to hematology-oncology due to progressive dyspnea, poor oral intake, and weight loss. He was diagnosed with relapsing polychondritis 2 years prior; however, his symptoms did not improve despite treatment. He was ultimately diagnosed with VEXAS syndrome with a mutation in UBA1 (ubiquitin-like modifier activating enzyme 1) and a concurrent SQSTM1 mutation. In addition, the coexistence of two mutations in the ubiquitination pathway in the same patient has not been reported to date. This patient and the treatment course were compared to pre-existing literature to increase awareness and improve the medical management of VEXAS syndrome.


Analysis of mammalian scrapie protein by novel monoclonal antibodies recognizing distinct prion protein glycoforms: an immunoblot and immunohistochemical study at the light and electron microscopic levels.
The availability of specific monoclonal antibodies (mAbs) recognizing the aberrant form (PrP(Sc)) of the cellular prion protein (PrP(C)) in different mammalian species is important for molecular diagnostics, PrP(Sc) typing and future immunotherapy. We obtained a panel of anti-PrP monoclonal antibodies in PrP(0/0) knock-out mice immunized with recombinant human PrP(23-231). Two mAbs, recognizing PrP epitopes in the alpha-helix 1 (mAb SA65) and alpha-helix 2 (mAb SA21) regions, immunoreacted with PrP(C) and PrP(Sc) and its proteolytic product, PrP27-30, from human, murine, bovine, caprine and ovine brains by Western blot. Remarkably, mAb SA21 recognized unglycosylated and monoglycosylated PrP with the second site occupied by glycan moieties, but not monoglycosylated PrP with the first consensus site occupied or highly glycosylated species. Immunoblots with mAb SA21 disclosed that PrP glycosylated at the second site accounted for the slower migrating form of the customary monoglycosylated PrP doublet. mAb SA65 immunolabelled all PrP glycoforms by Western blot and was highly efficient in detecting tissue PrP by immunohistochemistry in light microscopy and in immunoelectron microscopy. These novel anti-PrP mAbs provide tools to investigate the subcellular site of PrP deposition in mammalian prion diseases and may also contribute to assess the role of different PrP glycoforms in human and animal prion diseases.


The role of miR-130a-3p and SPOCK1 in tobacco exposed bronchial epithelial BEAS-2B transformed cells: Comparison to A549 and H1299 lung cancer cell lines.
In the pathogenesis of human lung cancer induced by tobacco smoke decreased expression levels of microRNAs (miRNAs) are known to occur. At present, the specific miRNAs expression levels reduced by tobacco smoke and subsequent lung cellular transformation remain to be determined. The aim of this study was thus to identify the miRNAs affected following cigarette-smoke exposure in bronchial epithelial BEAS-2B cells that were malignantly transformed into S30 cells. In addition, the miRNAs in S30 transformed cells were compared to human lung cancer cell lines A549 and H1299. Our results identified miR-130a-3p which was down-regulated in S30 cells as well as A549 and H1299 lung cancer cell lines. Using miRNA mimic, a correlation between elevated miR-130a-3p expression levels and reduced migration in A549 and H1299 cell lines and S30 cells was noted as evidenced by transwell and wound healing assays accompanied by enhanced apoptosis. Further, two online target genes prediction programs TargetScan and miRDB were employed to identify the miRNA target gene SPOCK1 in all three cell types. SPOCK1 expression was higher in unexposed bronchial epithelial BEAS-2B cells. It is of interest that however silencing SPOCK1 in transformed S30 cells exposed to cigarette-smoke a marked depression in cell migration was noted. Our findings demonstrate that upregulated miR-130a-3p was associated with reduced SPOCK1 expression in transformed S30 as well as lung cancer A549 and H1299 cell lines indicating that cigarette transformed cells behave similar to lung cancer cells and this process involves diminished lung cancer cells migration.


High-throughput genomic mapping of vector integration sites in gene therapy studies.
Gene therapy has enormous potential to treat a variety of infectious and genetic diseases. To date hundreds of patients worldwide have received hematopoietic cell products that have been gene-modified with retrovirus vectors carrying therapeutic transgenes, and many patients have been cured or demonstrated disease stabilization as a result (Adair et al., Sci Transl Med 4:133ra57, 2012; Biffi et al., Science 341:1233158, 2013; Aiuti et al., Science 341:1233151, 2013; Fischer et al., Gene 525:170-173, 2013). Unfortunately, for some patients the provirus integration dysregulated the expression of nearby genes leading to clonal outgrowth and, in some cases, cancer. Thus, the unwanted side effect of insertional mutagenesis has become a major concern for retrovirus gene therapy. The careful study of retrovirus integration sites (RIS) and the contribution of individual gene-modified clones to hematopoietic repopulating cells is of crucial importance for all gene therapy studies. Supporting this, the US Food and Drug Administration (FDA) has mandated the careful monitoring of RIS in all clinical trials of gene therapy. An invaluable method was developed: linear amplification mediated-polymerase chain reaction (LAM-PCR) capable of analyzing in vitro and complex in vivo samples, capturing valuable genomic information directly flanking the site of provirus integration. Linking this method and similar methods to high-throughput sequencing has now made possible an unprecedented understanding of the integration profile of various retrovirus vectors, and allows for sensitive monitoring of their safety. It also allows for a detailed comparison of improved safety-enhanced gene therapy vectors. An important readout of safety is the relative contribution of individual gene-modified repopulating clones. One limitation of LAM-PCR is that the ability to capture the relative contribution of individual clones is compromised because of the initial linear PCR common to all current methods. Here, we describe an improved protocol developed for efficient capture, sequencing, and analysis of RIS that preserves gene-modified clonal contribution information. We also discuss methods to assess dominant/overrepresented gene-modified clones in preclinical and clinical models.


Immunosuppression in viral oncogenesis. III. Effects of virus infection on interleukin 1 and interleukin 2 generation and responsiveness.
Malignant rabbit fibroma virus (MV) is an oncogenic immunosuppressive leporipoxvirus. We studied the effects of MV infection and MV-associated tumor-induced suppressor factor (TISF) on the production of and responsiveness to interleukins 1 and 2. Adherent cells from MV tumor-bearing rabbits elaborate adequate amounts of IL 1 in response to E. coli endotoxin. Neither live virus nor TISF alters the production or the responsiveness to IL 1. However, when we examined spleen cells from rabbits 7 days after MV inoculation, we noted that their ability to produce and respond to IL 2 is deficient. Despite their relatively poor capacity to produce IL 2, these spleen cells express receptor for IL 2 in normal amounts, as measured by the monoclonal antibody 7D4. TISF derived from T lymphocytes from MV tumor-bearing rabbits is by itself capable of inhibiting partially normal secretion of IL 2 and also the response of the cloned murine T cell line HT-2 to added IL 2. Full expression of the immunosuppressive capacity of spleen cells from MV tumor-bearing rabbits requires cell-cell contact, however, and cannot be replaced by either live virus or spleen cell supernatants. Such spleen cells inhibit normal mitogen responsiveness, a defect not remedied by adding exogenous IL 2. Immunologic dysfunction induced by MV infection is transient, and by 11 days after virus inoculation, actively mediated recovery from immunosuppression is observed. We found that spleen cells from rabbits studied 11 days postinoculation secreted IL 2 normally. Thus, immunologic dysfunction secondary to infection with malignant rabbit fibroma virus reflects deficiencies in both elaboration of and response to IL 2, and return of immune function later in the course of the infection is associated with return of the ability of lymphocytes to secrete IL 2.


The response of amphibian larvae to exposure to a glyphosate-based herbicide (Roundup WeatherMax) and nutrient enrichment in an ecosystem experiment.
Herbicides and fertilizers are widely used throughout the world and pose a threat to aquatic ecosystems. Using a replicated, whole ecosystem experiment in which 24 small wetlands were split in half with an impermeable barrier we tested whether exposure to a glyphosate-based herbicide, Roundup WeatherMax™, alone or in combination with nutrient enrichment has an effect on the survival, growth or development of amphibians. The herbicide was applied at one of two concentrations (low=210 μg a.e./L, high=2880 μg a.e./L) alone and in combination with nutrient enrichment to one side of wetlands and the other was left as an untreated control. Each treatment was replicated with six wetlands, and the experiment was repeated over two years. In the high glyphosate and nutrient enrichment treatment the survival of wood frog (Lithobates sylvaticus) larvae was lower in enclosures placed in situ on the treated sides than the control sides of wetlands. However, these results were not replicated in the second year of study and they were not observed in free swimming wood frog larvae in the wetlands. In all treatments, wood frog larvae on the treated sides of wetlands were slightly larger (<10%) than those on the control side, but no effect on development was observed. The most dramatic finding was that the abundance of green frog larvae (Lithobates clamitans) was higher on the treated sides than the control sides of wetlands in the herbicide and nutrient treatments during the second year of the study. The results observed in this field study indicate that caution is necessary when extrapolating results from artificial systems to predict effects in natural systems. In this experiment, the lack of toxicity to amphibian larvae was probably due to the fact the pH of the wetlands was relatively low and the presence of sediments and organic surfaces which would have mitigated the exposure duration.


Pb-induced changes in roots of two cultivated rice cultivars grown in lead-contaminated soil mediated by smoke.
Nowadays, public concerns regarding deleterious effect of lead (Pb) is on rise due to its abundance and toxic effect on plants and other living organisms. In plants, it has no noticeable biological importance but can cause various morphological, physiological, and biochemical malfunctions. To evaluate the remediating potential of plant-derived smoke (Cymbopogon jwarancusa), a pot culture experiment was designed to investigate the physiological, biochemical, metabolic, and antioxidant parameters of roots in lead (0 (control), 500, 1000, and 1500 ppm)-contaminated soil. Under dark condition, seeds were primed in smoke solution with two dilutions (1:500 and 1:1000) for 24 h. With an increasing concentration of Pb stress, fresh and dry weight and total nitrogen and protein contents decreased significantly while an increase was observed in smoke-treated seed. With increasing Pb stress level, metabolites (i.e., proline, total soluble sugar, total soluble protein, glycine betaine), and antioxidants (i.e., superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, malonyldialdehyde, and H2O2), contents of roots were increased in non-treated (without smoke treatment) samples, whereas comparatively, a low level of alteration in aforementioned metabolites and antioxidative parameters was observed in the seeds treated with smoke solution. These results suggest a positive role of smoke in alleviating lead-induced changes in roots of two cultivated cultivars of rice grown in Pb-contaminated soil.


Preventing Violence to Achieve Peaceful and Inclusive Societies.
This article is one in a series in which contributing authors discuss how the United Nations (UN) Sustainable Development Goals (SDGs) are linked to everyday clinical issues; national public health emergencies; and other nursing issues, such as leadership, shared governance, and advocacy. The 2030 Agenda for Sustainable Development, a 15-year plan of action to achieve the goals, was unanimously adopted by all UN member states in September 2015 and took effect on January 1, 2016. The Agenda consists of 17 SDGs addressing social, economic, and environmental determinants of health and 169 associated targets focused on five themes: people, planet, peace, prosperity, and partnership. The SDGs build on the work of the UN Millennium Development Goals, which were in effect from 2000 to 2015. The current article highlights SDG 16: "Promote peaceful and inclusive societies for sustainable development, provide access to justice for all, and build effective, accountable, and inclusive institutions at all levels."


[Modern data regarding the cardiovascular system lesions in patients with bronchopulmonary diseases and tuberculosis].
The article presents literature review on modern data regarding cardiovascular system lesions in patients with bronchopulmonary diseases and tuberculosis. To detect these changes is of great importance for an assessment of a functional condition of an organism as a whole.


Evaluation of the regenerative potential of 25% doxycycline-loaded biodegradable membrane vs biodegradable membrane alone in the treatment of human periodontal infrabony defects: a clinical and radiological study.
Microbial colonization of the barrier membranes used for guided tissue regeneration is inevitable and can lead to delayed healing. Antimicrobial coating of the membrane with 25% doxycycline paste has been attempted to prevent infection and achieve enhanced regeneration in periodontal infrabony defects. Twenty-four patients with 2-walled or 3-walled infrabony defects were selected and randomly divided into two equal groups. Infrabony defects of group A were treated with a biodegradable membrane coated with 25% doxycycline while those of group B were treated with membrane alone. Clinical assessment of probing depth and attachment level and radiographic evaluation of the defect depth was done preoperatively and at 12 and 24 weeks postoperatively. The relative efficacy of the two treatment modalities were evaluated using the paired Student's t- test and the comparative evaluation between the two groups was done using the independent Student's t -test. Both the groups exhibited a highly significant reduction in probing depth and gain in clinical attachment level and linear bone fill at the end of 24 weeks. Comparative evaluation between the two study groups revealed a significant reduction in probing depth ( P = 0.016 FNx01 ) and linear bone fill ( P = 0.02 FNx01 ) in group A as compared to group B. Mean gain in attachment level was greater for group A than for group B but the difference was statistically nonsignificant ( P = 0.065 NS ). The results suggest that doxycycline is beneficial in reducing membrane-associated infection and can potentiate regeneration through host modulation.


Optic nerve degeneration in a murine model of juvenile ceroid lipofuscinosis.
To investigate optic nerve degeneration associated with CLN3 deficiency in a murine model of juvenile neuronal ceroid lipofuscinosis (Batten disease). Using light and electron microscopy, the density and diameter of axons and the thickness of myelin in optic nerve were compared between age-matched cln3 knock-out (cln3-/-) and wild-type (129ev/TAC) mice. Western blot analysis was used to assay expression of Cln3 in mouse and primate retina and optic nerve. Morphologically identified mast cells were present in the meningeal sheaths surrounding the cln3-/- nerve and in the nerve itself. The cln3-/- optic nerve exhibited an overall loss of uniformity and integrity. Axon density in cln3-/- optic nerve was only 64% of that in wild-type optic nerve (P < 0.01). Accounting for differences in axon density, the diameter of axons in cln3-/- optic nerve was 1.2 times greater than in wild-type optic nerve (P < 0.01). Electron micrographs revealed large spaces between axons and 32% thinner myelin surrounding axons in cln3-/- mice than in wild type (P < 0.01). Western blot analysis demonstrated that Cln3 was expressed in retinas and optic nerves of mouse and primate. The presence of apparent mast cells in cln3-/- optic nerve suggests compromise of the blood-brain barrier. The absence of Cln3 causes loss of axons, axonal hypertrophy, and a reduction in myelination of retinal ganglion cells. Furthermore, expression of CLN3 in mouse and primate optic nerve links degeneration to loss of Cln3.


Divergent effects of obesity on fragility fractures.
Obesity was commonly thought to be advantageous for maintaining healthy bones due to the higher bone mineral density observed in overweight individuals. However, several recent studies have challenged the widespread belief that obesity is protective against fracture and have suggested that obesity is a risk factor for certain fractures. The effect of obesity on fracture risk is site-dependent, the risk being increased for some fractures (humerus, ankle, upper arm) and decreased for others (hip, pelvis, wrist). Moreover, the relationship between obesity and fracture may also vary by sex, age, and ethnicity. Risk factors for fracture in obese individuals appear to be similar to those in nonobese populations, although patterns of falling are particularly important in the obese. Research is needed to determine if and how visceral fat and metabolic complications of obesity (type 2 diabetes mellitus, insulin resistance, chronic inflammation, etc) are causally associated with bone status and fragility fracture risk. Vitamin D deficiency and hypogonadism may also influence fracture risk in obese individuals. Fracture algorithms such as FRAX(®) might be expected to underestimate fracture probability. Studies specifically designed to evaluate the antifracture efficacy of different drugs in obese patients are not available; however, literature data may suggest that in obese patients higher doses of the bisphosphonates might be required in order to maintain efficacy against nonvertebral fractures. Therefore, the search for better methods for the identification of fragility fracture risk in the growing population of adult and elderly subjects with obesity might be considered a clinical priority which could improve the prevention of fracture in obese individuals.


Pathways to Abortion at a Tertiary Care Hospital: Examining Obesity and Delays.
Advancing gestational age can increase the cost of an abortion and is a significant risk factor for complications. While obesity is not associated with increased risks, anecdotal evidence suggests that obese women seeking services at freestanding abortion clinics are often referred for hospital-based care, which can lead to delays. In 2016, a cross-sectional survey collected data on the experiences of 201 women who had obtained abortions at a hospital-based clinic in Philadelphia; rates of medical complications were determined from hospital records. Multivariable logistic regression analysis was used to assess if obesity was associated with whether patients had been referred from freestanding abortion clinics or reported other paths to care. Differences in wait time and up-front out-of-pocket costs were examined by women's referral status. No difference in rates of abortion complications was found between patient groups. Women who were severely obese (body mass index of at least 40 kg/m2 ) were more likely than normal-weight individuals to have been referred from a freestanding abortion clinic (odds ratio, 7.5). The median wait time to get an abortion was 28 days for referred patients and 12 days for others. Multivariable analysis confirmed that referred patients waited twice as long as other patients (rate ratio, 2.0) and paid 66% more in up-front costs. Future research is needed to determine whether obese women seeking abortions are being referred despite evidence that they do not require hospital-based care. If obese women are suffering delays because of referral, strategies to help overcome delay should also be explored.


Patterns of ischaemia and reperfusion in nipple-sparing mastectomy reconstruction with indocyanine green angiography.
Intraoperative assessment of mastectomy flaps and nipple-areola complex (NAC) with indocyanine green angiography (ICGA) for decision-making in delayed breast reconstruction after nipple-sparing mastectomy (NSM) remains to be fully elucidated. We evaluated patterns of ischaemia and reperfusion in NSM with delayed breast reconstruction and their outcomes. Single-institution retrospective study of delayed implant-based breast reconstructions following NSM due to poor perfusion analysis on ICGA. Intraoperative ICGA perfusion values and fluorescence patterns during the delayed and subsequent reconstruction operations were analysed. Fifty-six (45 patients) delayed breast reconstructions following NSM were performed. The median time to reconstruction was seven days (range, 4-21 days). A total of 112 fluorescence images were reviewed. Four patterns of ischaemia were identified during initial mastectomy (Type I, diffuse ischaemia; Type II, geographic ischaemia; Type III, incisional ischaemia; Type IV, NAC only ischaemia). All, but 1 breast, had adequate reperfusion during delayed reconstruction. Obesity (BMI ≥ 30) was associated with Type I ischaemia (p < 0.001). Mean ICGA absolute and relative perfusion values during initial mastectomy were significantly lower than the perfusion values during delayed reconstruction (absolute value 6.7 versus 40.2 units, p < 0.001; relative value 10% versus 44%, p < 0.001, respectively). There were no cases of partial-thickness or full-thickness necrosis. Delaying breast reconstruction for NSM with ischaemia predicted by ICGA may allow blood supply to the flap and NAC to improve, reducing the risk for necrosis. Distinct patterns of ischaemia and low perfusion values with ICGA may be used in the decision to delay reconstruction.


Intraoperative esophageal manometry in surgical treatment of achalasia: a reappraisal.
Intraoperative Esophageal Manometry is used during surgery for gastroesophageal reflux disease to obtain a better lower esophageal sphincter pressure, or during the Heller procedure for achalasia to verify the myotomy. The effectiveness of this manometry is not explicit. From 1977 to 1987, 58 patients underwent surgery for achalasia at our Institution. A transabdominal Heller's myotomy and a modified Dor's anterior fundoplication were performed in all cases. During the operation, Lower Esophageal Sphincter Pressure was measured in 38 patients (A group) to verify the completeness of the càr-diomyotomy; the fundoplication-induced pressure increase was also recorded. Twenty patients (B group) underwent surgery without intraoperative manometry. Forty-six patients (30 of A group and 16 of B group) were followed up for 24 to 120 months (median 68 months) by means of a clinical questionnaire, barium meal, manometry and 24-hour esophageal pH monitoring. In 21 cases, intraoperative manometry showed the completeness of the myotomy at the first record, while in the remaining 17 it revealed the persistence of a High Pressure Zone; in these, a more accurate incision of the muscle layers on the anterior surface of the esophagus was subsequently performed up to a complete abolition of the HPZ. Long-term results were excellent or good in 27 (90%) patients of A group and in 9 (56%) patients of B group. Dysphagia and gastro-esophageal reflux were the commonest findings in patients with poor results. The usefulness of intraoperative manometry in the surgical treatment of achalasia is emphasized and some technical details are reported.


Predictors of In-Hospital Mortality in Non-Elderly Adult Patients Emergently Admitted with Rhabdomyolysis: A 10-Year Analysis of 27,688 Patients from the National Inpatient Sample.
Rhabdomyolysis is a condition where muscle damage leads to the leakage of intracellular contents such as myoglobin and creatine kinase. These leak into systemic circulation and can cause detrimental effects. Due to the detrimental effect of rhabdomyolysis on patient mortality and potential complications, identifying factors that affect patient mortality in those with rhabdomyolysis could provide valuable insight in early management strategies and potentially benefit patient outcomes. The goal of this study was to identify independent predictors of in-hospital mortality in non-elderly adult patients who underwent emergency admission due to rhabdomyolysis. A retrospective cohort study was done by analyzing 27,688 non-elderly adult patients (18-64 years) with rhabdomyolysis who underwent emergency admission using the National Inpatient Sample (NIS) during 2005-2014. Factors such as demographic information, clinical course, and comorbidities were collected to identify predictors of in-hospital mortality. Chi square and student's t-tests were utilized to evaluate various group differences on categorical and continuous variables. Backward logistic regression analyses were performed to examine factors that could affect patient mortality. A total number of 27,688 non-elderly adult patients (age 18-64 years) were included, of which, 20,137 patients were male (72.8%) with a mean (SD) age of 40.60 (13.34) years, and 7,551 patients were female (27.3%) with a mean (SD) age of 45.63 (13.20) years. Multivariable backward logistic regression analysis was performed to evaluate the associations between mortality and different variables in our patient sample. Out of different factors, respiratory diseases, cardiac disease, and genitourinary system disease demonstrated the most significant association with mortality, shown by odds ratios of 3.67, 3.59, and 3.08, respectively. Additionally, patient age, history of surgical procedure, bacterial infection (other than tuberculosis), and cerebrovascular diseases were also positively associated with mortality. Their respective odds ratios were 1.03, 2.14, 2.13, and 2.66. Each additional year in age leads to 3% increased odds of mortality in non-elderly adult patients who are emergently admitted with rhabdomyolysis.


Effect of the orienting response on sensory discriminability.
The effects of orienting responses to different types of stimuli on acquisition of information were studied. Subjects underwent a standard habituation series of 15 trials. On Trial 16, they received one of following three stimuli: (a) no-change stimulus--same stimulus as habituation stimulus, (b) innocuous-change stimulus, (c) significant-change stimulus--the subject's own name. These orienting stimuli were followed 500 msec. later by an imperative stimulus (100 msec.) which contained a number of Chinese characters. The subjects were then unexpectedly asked to recall and recognize these characters. Recall and recognition were consistently superior in conditions with significant and innocuous change relative to no change, which indicated that the orienting response to unexpected stimuli is related to a generalized perceptual enhancement. The data pose problems for the position that the orienting response reflects only the passive 'registration' of the eliciting stimulus information. While digital pulse-amplitude data showed no difference among the conditions for the change trial, these conditions were differentiated by electrodermal and cardiac changes.


Inhibition of esophageal-carcinoma cell proliferation by genistein via suppression of JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways.
Esophageal carcinoma (EsC) is a clinically challenging neoplastic disease. Genistein, a natural isoflavone product, has anti-tumor properties. Through in vitro and in vivo studies, we found that genistein suppressed EsC cell proliferation in a time- and concentration-dependent manner. In addition, genistein markedly promoted apoptosis and arrested cell cycle at the G0/G1 phase in a concentration-dependent manner. Furthermore, high concentrations of genistein have no adverse effect on normal esophageal epithelial cells. Mechanistically, genistein treatment strikingly reduced the expression of cell cycle-associated genes, and up-regulated the expression of cell apoptosis-related genes in EsC cells. Additionally, genistein dramatically decreased epidermal growth factor receptor (EGFR) expression and attenuated its down-stream signaling molecules STAT3, MDM2, Akt and JAK1/2 phosphorylation, resulting in inhibited nuclear translocation of STAT3 and MDM2, thereby inhibiting the JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways. In xenograft nude mice, genistein administration strikingly impaired tumor growth in a dose-dependent manner. Moreover, similar disturbances in molecular mechanisms were observed in vivo. Taken together, genistein suppressed the JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways by decreasing EGFR expression, leading to cell apoptosis, cell cycle arrest, and proliferation inhibition in EsC cells. Our findings suggest that genistein may be a promising alternative adjuvant therapy for patients with EsC.


Taurine ameliorates cholesterol metabolism by stimulating bile acid production in high-cholesterol-fed rats.
This study was designed to investigate the effects of dietary taurine on cholesterol metabolism in high-cholesterol-fed rats. Male Sprague-Dawley rats were randomly divided into two dietary groups (n = 6 in each group): a high-cholesterol diet containing 0.5% cholesterol and 0.15% sodium cholate, and a high-cholesterol diet with 5% (w/w) taurine. The experimental diets were given for 2 weeks. Taurine supplementation reduced the serum and hepatic cholesterol levels by 37% and 32%, respectively. Faecal excretion of bile acids was significantly increased in taurine-treated rats, compared with untreated rats. Biliary bile acid concentrations were also increased by taurine. Taurine supplementation increased taurine-conjugated bile acids by 61% and decreased glycine-conjugated bile acids by 53%, resulting in a significant decrease in the glycine/taurine (G/T) ratio. Among the taurine-conjugated bile acids, cholic acid and deoxycholic acid were significantly increased. In the liver, taurine supplementation increased the mRNA expression and enzymatic activity of hepatic cholesterol 7α-hydroxylase (CYP7A1), the rate-limiting enzyme for bile acid synthesis, by three- and two-fold, respectively. Taurine also decreased the enzymatic activity of acyl-CoA:cholesterol acyltransferase (ACAT) and microsomal triglyceride transfer protein (MTP). These observations suggest that taurine supplementation increases the synthesis and excretion of taurine-conjugated bile acids and stimulates the catabolism of cholesterol to bile acid by elevating the expression and activity of CYP7A1. This may reduce cholesterol esterification and lipoprotein assembly for very low density lipoprotein (VLDL) secretion, leading to reductions in the serum and hepatic cholesterol levels.


Rates and patterns of recurrence after curative intent resection for gastric cancer: a United States multi-institutional analysis.
Reports on recurrence and outcomes of US patients with gastric cancer are scarce. The aim of this study was to determine incidence and pattern of recurrence after curative intent surgery for gastric cancer. Using the multi-institutional US Gastric Cancer Collaborative database, we identified 817 patients undergoing curative intent resection for gastric cancer between 2000 and 2012. Patterns and rates of recurrence along with associated risk factors were identified using adjusted regression analysis. Recurrences were classified as locoregional, peritoneal, or hematogenous. Median patient age was 65.8 years (interquartile range [IQR] 56.4, 74.7); the majority of patients were male (n = 462, 56.6%) and white (n = 511, 62.5%). At the time of surgery, the majority of patients underwent a partial gastrectomy (n = 481, 59.2%) with a complete R0 resection achieved in 91.6% (n = 748) of patients. At the time of last follow-up, 244 (29.9%) of 817 patients developed a recurrence; 163 (66.8%) patients had recurrence at only a single site; the remaining 81 (33.2%) had multiple sites of initial recurrence. Among patients who recurred at a single site, recurrence was most common at a distant location and included hematogenous (n = 57, 23.4%) or peritoneal (n = 47, 19.3%) only metastasis. Tumors at the gastroesophageal junction (odds ratio [OR] 3.18, 95% CI 1.08 to 9.40; p = 0.04) were associated with higher risk of locoregional recurrence, while the presence of multiple lesions (OR 10.82, 95% CI 3.56 to 32.85; p < 0.001) remained associated with an increased risk of distant hematogenous recurrence after adjusted analysis. Recurrence was associated with worse survival, with a median recurrence-free survival of 10.8 months (IQR 8.9, 12.8) among those who experienced a recurrence. Nearly one-third of patients experienced recurrence after gastric cancer surgery. The most common site of recurrence was distant.


Traumatic brain injury, posttraumatic stress disorder, and pain diagnoses in OIF/OEF/OND Veterans.
To identify the prevalence of traumatic brain injury (TBI), posttraumatic stress disorder (PTSD), and pain in Veterans from Operation Iraqi Freedom/Operation Enduring Freedom/Operation New Dawn (OIF/OEF/OND), Veterans who received any inpatient or outpatient care from Veterans Health Administration (VHA) facilities from 2009 to 2011 were studied. A subset of Veterans was identified who were diagnosed with TBI, PTSD, and/or pain (head, neck, or back) as determined by their International Classification of Diseases-9th Revision-Clinical Modification codes. Between fiscal years 2009 and 2011, 613,391 Veterans accessed VHA services at least once (age: 31.9 +/- 9.6 yr). TBI diagnosis in any 1 year was slightly less than 7%. When data from 3 years were pooled, 9.6% were diagnosed with TBI, 29.3% were diagnosed with PTSD, and 40.2% were diagnosed with pain. The full polytrauma triad expression (TBI, PTSD, and pain) was diagnosed in 6.0%. Results show that increasing numbers of Veterans from OIF/OEF/OND accessed VHA over a 3 year period. Among those with a TBI diagnosis, the majority also had a mental health disorder, with approximately half having both PTSD and pain. While the absolute number of Veterans increased by over 40% from 2009 to 2011, the proportion of Veterans diagnosed with TBI and the high rate of comorbid PTSD and pain in this population remained relatively stable.


Recommended standardized anatomic terminology of the posterior female pelvis and vulva based on a structured medical literature review.
Anatomic terminology in both written and verbal forms has been shown to be inaccurate and imprecise. Here, we aimed to (1) review published anatomic terminology as it relates to the posterior female pelvis, posterior vagina, and vulva; (2) compare these terms to "Terminologia Anatomica," the internationally standardized terminology; and (3) compile standardized anatomic terms for improved communication and understanding. From inception of the study to April 6, 2018, MEDLINE database was used to search for 40 terms relevant to the posterior female pelvis and vulvar anatomy. Furthermore, 11 investigators reviewed identified abstracts and selected those reporting on posterior female pelvic and vulvar anatomy for full-text review. In addition, 11 textbook chapters were included in the study. Definitions of all pertinent anatomic terms were extracted for review. Overall, 486 anatomic terms were identified describing the vulva and posterior female pelvic anatomy, including the posterior vagina. "Terminologia Anatomica" has previously accepted 186 of these terms. Based on this literature review, we proposed the adoption of 11 new standardized anatomic terms, including 6 regional terms (anal sphincter complex, anorectum, genital-crural fold, interlabial sulcus, posterior vaginal compartment, and sacrospinous-coccygeus complex), 4 structural terms (greater vestibular duct, anal cushions, nerve to the levator ani, and labial fat pad), and 1 anatomic space (deep postanal space). In addition, the currently accepted term rectovaginal fascia or septum was identified as controversial and requires further research and definition before continued acceptance or rejection in medical communication. This study highlighted the variability in the anatomic nomenclature used in describing the posterior female pelvis and vulva. Therefore, we recommended the use of standardized terminology to improve communication and education across medical and anatomic disciplines.


"Smart" Triiodide Compounds: Does Halogen Bonding Influence Antimicrobial Activities?
Antimicrobial agents containing symmetrical triiodides complexes with halogen bonding may release free iodine molecules in a controlled manner. This happens due to interactions with the plasma membrane of microorganisms which lead to changes in the structure of the triiodide anion. To verify this hypothesis, the triiodide complex [Na(12-crown-4)2]I3 was prepared by an optimized one-pot synthesis and tested against 18 clinical isolates, 10 reference strains of pathogens and five antibiotics. The antimicrobial activities of this symmetrical triiodide complex were determined by zone of inhibition plate studies through disc- and agar-well-diffusion methods. The triiodide complex proved to be a broad spectrum microbicidal agent. The biological activities were related to the calculated partition coefficient (octanol/water). The microstructural analysis of SEM and EDS undermined the purity of the triiodide complex. The anionic structure consists of isolated, symmetrical triiodide anions [I-I-I]- with halogen bonding. Computational methods were used to calculate the energy required to release iodine from [I-I-I]- and [I-I···I]-. The halogen bonding in the triiodide ion reduces the antibacterial activities in comparison to the inhibitory actions of pure iodine but increases the long term stability of [Na(12-crown-4)2]I3.


The need for regional planning of perinatal care.
Regionalisation is urgently needed to provide the best possible maternal, fetal and newborn care. In practice this means organising perinatal care of three specified levels and referring the high-risk problems to the regional centre, preferably before labour and delivery. The programme requires careful planning and constant teamwork.