Hormonal modulation of the accumulation of 5-aminolevulinic acid-induced protoporphyrin and phototoxicity in prostate cancer cells.
The effect of hormonal modulation on the response of human prostate cancer cell lines to photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) was studied. Two cell lines, one responsive to androgens (LNCaP) and the other non-responsive (PC-3), were used. Fetal bovine serum was depleted of steroid hormones by stripping with charcoal-dextran, and then resupplied with known concentrations of the androgen 5alpha-dihydrotestosterone (DHT) or the estrogen estradiol. It was found that LNCaP cells alone increased growth rate in response to both added hormones, but only androgen had an effect on PDT treatment. LNCaP cells pretreated with 10(-7) M DHT and then 1 mM ALA accumulated 70% more PpIX, compared to cells pre-treated with 10(-12) M DHT. Exposure of the cells treated with high DHT and ALA to 630 nm light led to an 85% decrease in the number of surviving cells compared to the low DHT treated group. The uptake of 14C-ALA was increased with high DHT treatment of the cells, consistent with the above data. No effect of hormones was seen with PC-3 cells or with either cell line and the exogenous photosensitizer benzoporphyrin derivative.


Differential effects of dentate kindling on working and reference spatial memory in the rat.
An 8-arm radial maze was used to examine the effects of dentate kindling on the performance of a task which requires both working (short-term) and reference (long-term) memory. During the 4-week post-operative training period, control and experimental rats performed both components of the task equally well. Performance of the reference memory component, but not the working memory component of the task was significantly impaired during kindling stimulation. The impairment in reference memory appeared to be primarily due to the elicitation of afterdischarges, since the disruption in maze performance was evident in the pre-convulsive stages of kindling. Following the cessation of kindling stimulation, a gradual recovery in reference memory performance was observed. Dentate kindling may provide a useful model for the study of long-term memory deficits associated with temporal lobe epilepsy.


An intuitive surgical handle design for robotic neurosurgery.
The expanded endoscopic endonasal approach, a representative example of keyhole brain surgery, allows access to the pituitary gland and surrounding areas through the nasal and sphenoid cavities. Manipulating rigid instruments through these constrained spaces makes this approach technically challenging, and thus, a handheld robotic instrument could expand the surgeon's capabilities. In this study, we present an intuitive handle prototype for such a robotic instrument. We have designed and fabricated a surgical instrument handle prototype that maps the surgeon's wrist directly to the robot joints. To alleviate the surgeon's wrist of any excessive strain and fatigue, the tool is mounted on the surgeon's forearm, making it parallel with the instrument's shaft. To evaluate the handle's performance and limitations, we constructed a surgical task simulator and compared our novel handle with a standard neurosurgical tool, with the tasks being performed by a consultant neurosurgeon. While using the proposed handle, the surgeon's average success rate was [Formula: see text], compared to [Formula: see text] when using a conventional tool. Additionally, the surgeon's body posture while using the suggested prototype was deemed acceptable by the Rapid Upper Limb Assessment ergonomic survey, while early results indicate the absence of a learning curve. Based on these preliminary results, the proposed handle prototype could offer an improvement over current neurosurgical tools and procedural ergonomics. By redirecting forces applied during the procedure to the forearm of the surgeon, and allowing for intuitive surgeon wrist to robot-joints movement mapping without compromising the robotic end effector's expanded workspace, we believe that this handle could prove a substantial step toward improved neurosurgical instrumentation.


Genome-wide identification, structure analysis and expression profiling of phospholipases D under hormone and abiotic stress treatment in chickpea (Cicer arietinum).
Phospholipases D (PLDs) are phospholipid hydrolyzing enzymes and crucial components of lipid signaling in plants. PLDs are implicated in stress responses in different plants however, characterization of PLDs in chickpea is missing. Here, we identify 13 PLD genes in the chickpea genome. PLD family could be divided into α, β, δ, ε and ζ isoforms based on sequence and structure. Protein remodeling described that chickpea PLDs are composed of defined arrangements of α-helix, β-sheets and short loops. Phylogenetic analysis suggested evolutionary conservation of chickpea PLD family with dicots. In-planta subcellular localization showed the plasma membrane localization of chickpea PLDs. All PLD promoters had hormone and stress related cis-regulatory elements, which suggested overlapping function of PLDs in hormone and abiotic stress signaling. The qRT-PCR expression analysis revealed that most PLD genes are differentially expressed in multiple abiotic stresses (drought, salt and cold stress). Moreover, several PLD genes had overlapping expression in abiotic stress and ABA and JA treatment. These observations indicate the involvement of PLD gene family in cross-talk of phytohormone and abiotic stress signaling in chickpea. Thus, present study opens new avenues of utilizing PLD related information for understanding hormone-regulated abiotic stress signaling in legume crops.


Tumor Necrosis Factor Alpha Overexpression Induces Mainly Osteoclastogenesis at the Vertebral Site.
Syndesmophyte occurrence and axial bone loss were investigated in the heterozygous Tg187 tumor necrosis factor (TNF) transgenic mouse model (Tg-huTNF) of arthritis. Female and male Tg-huTNF mice were compared to wild-type mice (WT) at 2, 4, 6, 8, and 10 weeks. Syndesmophytes, intervertebral disc space, osteoclasts, osteoid surface, and vertebra microarchitecture were assessed by histomorphometry and microcomputed tomography. No spontaneous syndesmophyte formation was detected in Tg-huTNF compared to WT mice. However, increased porosity was observed mainly in peridiscal lumbar vertebra. Accordingly, bone microarchitecture parameters were altered in Tg-huTNF mice, with decrease in bone volume fraction, and trabecular number and thickness after 6 weeks compared to WT (p < 0.05). Osteoclast count and surface were increased (p < 0.01). Moreover, the non-mineralized (osteoid) surface was also increased in Tg-huTNF after 6 weeks (p < 0.01). Despite increased osteoclast and osteoid surfaces, an imbalance between both was observed in favour of osteoid surface at the early phase and then to osteoclast surface. These results demonstrated an axial bone loss in the Tg-huTNF model, additional to the common limb arthritis, related to overexpression of TNF. However, the absence of syndesmophyte and the increase of osteoid surface suggested that chronic inflammation might block bone mineralisation. Finally, the relative increased osteoid surface was not enough to compensate the high osteoclast activity.


Microsurgical practice on avulsed skin.
The use of avulsed skin is recommended as a source of vessels for trainees to practise microvascular techniques. If it cannot be used immediately it can be stored in the refrigerator for later use.


Epidemics, pandemics and income inequality.
The novel coronavirus is part of a series of infectious disease outbreaks that include: Ebola, Avian influenza, Middle East respiratory syndrome coronavirus, and Influenza A. This paper addresses the question of how do these epidemics and pandemics affect income inequality in countries around the world during the first two decades of this century. To achieve its objective, the paper develops a model that indicates a positive association between these health crises and income inequality. To empirically test our theoretical predictions, the paper explores the effect on the Gini coefficient of a dummy variable that indicates the occurrence of an epidemic or a pandemic in a country in a given year and the number of deaths per 100,000. To properly address potential endogeneity, we implement a Three-Stage-Least Squares technique. The estimation shows that the number of deaths per 100,000 population variable has a statistically significant positive effect on the Gini coefficient, especially when we incorporate COVID-19 data. This suggests that not only the occurrence, but also the health consequences of COVID-19 have a significant and economically important effect on income inequality. The purpose of the study is to examine the effect of epidemics and pandemics on income inequality. This has important implications as the outcome of this study can guide policymakers into implementing policies that can mitigate the economic consequences of these health crises. The study is a cross country analysis using fixed effects estimation. To address potential endogeneity and determine causality, the paper uses the Three-Stage-Least-Squares estimation. The paper finds that the number of epidemic deaths per 100,000 population variable has a statistically significant positive effect on the Gini coefficient, especially when we incorporate COVID-19 data. The paper finds that it is not only the occurrence of an epidemic, captured by the epidemics dummy variable, but also the health consequences, captured by the number of deaths per 100,000 population, that have a significant effect on income inequality. This is especially the case when we incorporate COVID-19 in our analysis. Not Applicable.


An efficient method of antibody elution for the successive or simultaneous localization of two antigens by immunocytochemistry.
The method described in this paper is available for removal of antibodies retained by tissue antigen after immunohistochemical staining. Its application to the antehypophysis has allowed the successive or the simultaneous localization of two different hormones.


Same-day thyroidectomy: a review of practice patterns and outcomes for 1,168 procedures in New York State.
There has been a shift of procedures from the inpatient to the outpatient setting. Same-day thyroidectomy (SDT) has been reported in high-volume single-institution series, but few studies have evaluated its widespread use. Patients undergoing thyroidectomy for benign and malignant thyroid disease were abstracted from the 2004 New York State inpatient (SID) and ambulatory surgery (SASD) databases. SDTs were discharged on the same day as their surgery. Patient and provider (surgeon and hospital volume) characteristics were associated with outcomes, including probability of SDT versus hospital admission and 30-day rehospitalization, by bivariate and multivariate analyses. A total of 6,762 thyroidectomies were identified; 17% (1,168) were SDTs. Patients undergoing SDT compared to thyroidectomy with admission were more often white (80 vs. 65%, P < 0.001), with private insurance (80 vs. 70%, P < 0.001) and fewer comorbidities (96 vs. 89% with Charlson scores of none/low, P < 0.001). SDT was performed more often by high-volume surgeons (48 vs. 31%, P < 0.001) and at high-volume hospitals (61 vs. 35%, P < 0.001). Rehospitalization rates of 1.4 and 2.4% were observed for SDT and inpatient thyroidectomy, respectively (P = NS). In multivariate analysis, thyroidectomy by a high-volume surgeon was associated with a higher chance of same-day discharge (odds ratio = 2.3, P < 0.001). Nearly 20% of thyroidectomy patients undergo SDT in New York State. They have different demographic and clinical characteristics than patients undergoing thyroidectomy who are admitted. There seem to be a few high-volume surgeons and centers with extensive SDT experience. More research is needed to explore optimized patient triage and patterns of referral to centers of excellence.


Lack of Annexin A6 Exacerbates Liver Dysfunction and Reduces Lifespan of Niemann-Pick Type C Protein-Deficient Mice.
Niemann-Pick type C (NPC) disease is a lysosomal storage disorder characterized by cholesterol accumulation caused by loss-of-function mutations in the Npc1 gene. NPC disease primarily affects the brain, causing neuronal damage and affecting motor coordination. In addition, considerable liver malfunction in NPC disease is common. Recently, we found that the depletion of annexin A6 (ANXA6), which is most abundant in the liver and involved in cholesterol transport, ameliorated cholesterol accumulation in Npc1 mutant cells. To evaluate the potential contribution of ANXA6 in the progression of NPC disease, double-knockout mice (Npc1-/-/Anxa6-/-) were generated and examined for lifespan, neurologic and hepatic functions, as well as liver histology and ultrastructure. Interestingly, lack of ANXA6 in NPC1-deficient animals did not prevent the cerebellar degeneration phenotype, but further deteriorated their compromised hepatic functions and reduced their lifespan. Moreover, livers of Npc1-/-/Anxa6-/- mice contained a significantly elevated number of foam cells congesting the sinusoidal space, a feature commonly associated with inflammation. We hypothesize that ANXA6 deficiency in Npc1-/- mice not only does not reverse neurologic and motor dysfunction, but further worsens overall liver function, exacerbating hepatic failure in NPC disease.


Evaluation of modified Estimation of Physiologic Ability and Surgical Stress in gastric carcinoma surgery.
We recently modified our prediction scoring system "Estimation of Physiologic Ability and Surgical Stress" and have designated the current version mE-PASS. This scoring system has been designed to obtain predicted postoperative mortality rates before surgery and this study was performed to assess its usefulness in elective surgery for gastric carcinoma. We investigated seven variables for mE-PASS and evaluated the postoperative course in 3,449 patients who underwent elective surgery for gastric carcinoma in Japan between August 20, 1987 and April 9, 2007, in order to quantify the predicted in-hospital mortality rates (R). The calibration and discrimination power of R were assessed using the Hosmer-Lemeshow test and the area under the receiver operating characteristic curve (AUC), respectively. The ratios of observed-to-estimated mortality rates (OE ratios) were quantified as a measure of quality. The overall postoperative morbidity and mortality rates were 19.0 and 2.0%, respectively. R demonstrated good power in calibration (χ(2) value, 12.5; df 8; P = 0.89) as well as discrimination (AUC, 95% confidence intervals: 0.80, 0.75-0.85). The OE ratios between hospitals ranged from 0.44 to 1.8. Overall, the OE ratios seemed to improve with time (OE ratio, 95% confidence intervals: 1.3, 0.73-2.4 for the early period between 1987 and 2000; 1.0, 0.59-1.7 for the middle period between 2001 and 2004; and 0.65, 0.36-1.2 for the late period between 2005 and 2007). Based on these findings, mE-PASS might be useful for medical decision-making and for assessing the quality of care in elective surgery for gastric carcinoma.


Acute infection with Toxoplasma gondii oocysts preferentially activates non-neuronal cells expressing serotonin in the jejunum of rats.
The interaction of Toxoplasma gondii with the gastrointestinal tract of its host is highly regulated. Once ingested, the parasite crosses the epithelium without altering the permeability of the intestinal barrier. Nevertheless, many studies report alterations ranging from structural to functional damage in cells and tissues that make up the wall of the small and large intestine. Although the immune response to the parasite has been extensively studied, the role of serotonin (5-HT) in toxoplasmosis is poorly understood. Here we investigate the distribution of cells expressing 5-HT and its effects on cells and tissues of the jejunal wall of rats after 2, 3, or 7 days of T. gondii infection. KEY RESULTS: Our results show that transposition of the jejunal epithelium by T. gondii leads to ruptures in the basement membrane and activation of the immune system, as confirmed by the decrease in laminin immunostaining and the increase in the number of mast cells, respectively. CONCLUSIONS AND INFERENCES: We showed an increase in the number of enterochromaffin cells and mast cells expressing 5-HT in the jejunal wall. We also observed that the percentage of serotonergic mast cells increased in the total population. Thus, we can suggest that oral infection by T. gondii oocysts preferentially activates non-neuronal cells expressing 5-HT. Together, these results may explain both the changes in the extracellular matrix and the morphology of the enteric ganglia.


Differences in gene expression in field populations of Wolbachia-infected Aedes aegypti mosquitoes with varying release histories in northern Australia.
Aedes aegypti is the principal mosquito vector of dengue, yellow fever, Zika and chikungunya viruses. The wMel strain of the endosymbiotic bacteria Wolbachia pipientis was introduced into the vector as a novel biocontrol strategy to stop transmission of these viruses. Mosquitoes with Wolbachia have been released in the field in Northern Queensland, Australia since 2011, at various locations and over several years, with populations remaining stably infected. Wolbachia infection is known to alter gene expression in its mosquito host, but whether (and how) this changes over the long-term in the context of field releases remains unknown. We sampled mosquitoes from Wolbachia-infected populations with three different release histories along a time gradient and performed RNA-seq to investigate gene expression changes in the insect host. We observed a significant impact on gene expression in Wolbachia-infected mosquitoes versus uninfected controls. Fewer genes had significantly upregulated expression in mosquitoes from the older releases (512 and 486 from the 2011 and 2013/14 release years, respectively) versus the more recent releases (1154 from the 2017 release year). Nonetheless, a fundamental signature of Wolbachia infection on host gene expression was observed across all releases, comprising upregulation of immunity (e.g. leucine-rich repeats, CLIPs) and metabolism (e.g. lipid metabolism, iron transport) genes. There was limited downregulation of gene expression in mosquitoes from the older releases (84 and 71 genes from the 2011 and 2013/14 release years, respectively), but significantly more in the most recent release (509 from the 2017 release year). Our findings indicate that at > 8 years post-introgression into field populations, Wolbachia continues to profoundly impact expression of host genes, such as those involved in insect immune response and metabolism. If Wolbachia-mediated virus blocking is underpinned by these differential gene expression changes, our results suggest it may remain stable long-term.


Iodine monitoring models contribute to avoid adverse birth outcomes related more than adequate iodine intake.
Iodine plays an important role in pregnancy. How to maintain adequate iodine intake amongst pregnant women in each trimester of pregnancy to prevent adverse birth outcomes in central China is a challenge for clinical practice. 870 pregnant women and their infants were enrolled in the study. Urinary iodine concentration (UIC) was measured using an inductively coupled plasma mass spectrometry (ICP-MS). Maternal and newborn information were obtained during follow-up. Multinomial logistic regression models were established. Median UIC of pregnant women was 172 ± 135 μg/L which is currently considered to be sufficient. Multivitamin supplements containing iodine, iodized salt intake and frequent milk intake were significantly associated with higher UIC. Multivariate logistic regression analysis showed that multivitamin supplements containing iodine and milk consumption were risk factors for more than adequate iodine (UIC ≥ 250 μg/L). Iodine-rich diet was significantly related to heavier birthweight, larger head circumference and longer femur length of the newborns while more than adequate iodine intake (UIC ≥ 250 μg/L) was a risk factor for macrosomia. Logistic regression models based on potential risk factors involving iodine containing supplements and iodine-rich diet were established to predict and screen pregnant women with high risk of more than adequate iodine intake among local pregnant women in different trimesters and guide them to supplement iodine reasonably to prevent the risk. Multivitamin supplements containing iodine and milk consumption were risk factors for maternal UIC ≥ 250 μg/L which was a risk factor for macrosomia. Iodine monitoring models were established to provide guidance for pregnant women to reduce the risk of more than adequate iodine intake, thereby contributing to reduce the risk of having a macrosomia.


Value of pre-operative serum CA125 level for prediction of prognosis in patients with endometrial cancer.
High pre-operative CA125 levels in women with endometrial cancer may be related to lymph node metastases and poor prognosis. To evaluate whether pre-operative cancer antigen 125 (CA125) levels are associated with lymph node metastases and prognosis in endometrial cancer. One hundred and twenty women with endometrial cancer were retrospectively reviewed for pre-operative CA125 levels. The results were then correlated with the clinicopathological outcome. An elevated CA125 (>40 U/mL) was significantly correlated with higher stage, higher grade, increased depth of myometrial invasion, lymph node metastases and the presence of lympho-vascular space involvement in endometrial cancer. Five-year overall survival (OS) and recurrence-free survival (RFS) rates were significantly higher in women with endometrial cancer with CA125 ≤ 40 U/mL than those with CA125 > 40 U/mL (P < 0.001). When women were further stratified according to CA125 levels and lymph node status, OS and RFS were highest for those with CA125 ≤ 40 U/mL and without lymph node metastases, and lowest for those with lymph node metastases and CA125 > 40 U/mL (P < 0.001). The testing of pre-operative CA125 levels may a useful prognostic tool in endometrial cancer management.


Transmission of methicillin-resistant Staphylococcus pseudintermedius between infected dogs and cats and contact pets, humans and the environment in households and veterinary clinics.
The objective of this study was to investigate the prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in people, pets and the environment in households with a pet with a clinical MRSP-infection within the past year. Personnel and the environment at veterinary clinics were also screened. Nasal swabs (humans), nasal and perineal swabs (pets) and environmental wipes were examined using selective culturing. Twenty households were enrolled; 10/20 index cases still had clinical signs of infection at the start of the study and all were MRSP-positive. Of the remaining 10 index cases five were MRSP-positive in nasal and/or perineal samples. Five of 14 (36%) contact dogs and four of 13 (31%) contact cats were found MRSP-positive. In the households with an index case with clinical signs of infection 6/7 (86%) contact animals were MRSP-positive. MRSP was cultured from 2/45 (4%) human nasal samples. Domestic contamination was widespread as positive samples were found in 70% of the households and 44% of all environmental samples were MRSP-positive. In all but one of these MRSP-positive households the index case was still MRSP positive. Among the personnel in veterinary clinics 4/141 (3%) were MRSP-positive. MRSP was cultured from 31/200 environmental samples in 7/13 clinics at the first sampling and in 3/6 clinics the environment remained MRSP-positive after cleaning and disinfection indicating that current cleaning procedures often were unable to eliminate MRSP. These results show that transmission of MRSP between infected or colonized dogs and cats and healthy people does occur but is relatively uncommon, while transmission to contact pets occurs frequently, especially when the index case still has clinical signs of MRSP-infection.


Scedosporium apiospermum mycetoma with bone involvement successfully treated with voriconazole.
Treatment of Scedosporium apiospermum mycetoma usually requires limb amputation. A 49-year-old woman, from Ivory Coast, was diagnosed with Madura foot in 1995. She failed to respond to several treatments including itraconazole, fluconazole and co-trimoxazole, and refused limb amputation. In December 2002 she was admitted to hospital in France with a painful, swollen right leg and foot. She had no fever and C-reactive protein was 120 mg/l. Magnetic resonance imaging (MRI) confirmed the destruction of tarsus bones with a tibia extension. Voriconazole (400 mg/day) treatment was initiated in March 2003; a significant clinical improvement was observed within 4 months as confirmed by C-reactive protein (16 mg/l) and MRI. Voriconazole was maintained for 18 months with good tolerance. Cholestasis appeared after the first month and remained stable. In October 2004 voriconazole was discontinued due to side effects on the liver (alanine aminotransferase 17 times the normal level); MRI showed impressive regression of bone lesions. As of July 2005, the patient remains clinically well. Voriconazole appears to be a promising drug for the treatment of S. apiospermum mycetomas.


Factors Associated With Discharge Destination in Older Patients: Finnish Community Hospital Cohort Study.
Primary care physician-led community hospitals provide basic hospital care for older people in Finland. Yet little is known of the outcomes of the care. We investigated factors associated with discharge destination after hospitalization in a community hospital and the role of active rehabilitation during the stay. Prospective observational study. Short-term community hospital stays of older adults (≥65 years) living in the Kuopio University Hospital district in central and eastern Finland. Data on short-term (1-31 days) hospital stays from 51 community hospitals were collected with an electronic survey between January and June 2016. Physicians, secretaries, and rehabilitation staff from each community hospital completed the data collection form. Discharge destination was defined as home, residential care or death, and active rehabilitation as frequency of rehabilitation at least once a day. Analyses were conducted using the Bayesian approach and the BayesiaLab 9.1 tool. Data of 11,628 community hospital stays were analyzed. The patients' mean age was 81.6 years (SD 7.9), and 57.5% were women. A younger age (65-74 years), a high number of rehabilitation staff (>2 per 10 patients), and receiving rehabilitation at least once a day were associated with discharging patients to their own homes. Daily rehabilitation was associated with returning to home in all patient groups. Older patients admitted to a community hospital for any reason may benefit from active rehabilitation. The role of community hospitals in the acute care and rehabilitation of older patients is important in aging societies.


Positron emission tomography with 11C-flumazenil in the rat shows preservation of binding sites during the acute phase after 2 h-transient focal ischemia.
Positron emission tomography (PET) studies in humans have used (11)C-flumazenil (FMZ) to assess neuronal viability after stroke. Here we aimed to study whether (11)C-FMZ binding was sensitive to neuronal damage in the acute phase following ischemia/reperfusion in the rat brain. Transient (2 h followed by reperfusion) and permanent intraluminal middle cerebral artery occlusion was carried out. (11)C-FMZ binding was studied by PET up to 24 h after the onset of ischemia. Tissue infarction was evaluated post-mortem at 24 h. Immunohistochemistry against a neuronal nuclei specific protein (NeuN) was performed to assess neuronal injury. No decrease in (11)C-FMZ binding was detected in the ipsilateral cortex up to 24 h post-ischemia in the model of transient occlusion despite the fact that rats developed cortical and striatal infarction, and neuronal injury was clearly apparent at this time. In contrast, (11)C-FMZ binding was significantly depressed in the ipsilateral cortex at 24 h following permanent ischemia. This finding evidences that (11)C-FMZ binding is not sensitive to neuronal damage on the acute phase of ischemia/reperfusion in the rat brain.


Talpid3-Mediated Centrosome Integrity Restrains Neural Progenitor Delamination to Sustain Neurogenesis by Stabilizing Adherens Junctions.
Neurogenesis in the developing neocortex relies on extensive mitosis of radial glial cells (RGCs) in the apical surface. The nuclear migration of epithelial-like RGCs is fundamentally important for proper mitosis, but how the apical processes of RGCs are anchored to ensure the nucleokinetic behavior of RGCs remains unclear. Here we find that Talpid3, related to Joubert syndrome, is localized to the mother centriole of RGCs and is required for their apical mitosis. Genetic silencing of Talpid3 causes abnormal RGC delamination and thereby impairs their interkinetic nuclear migration in both cell-autonomous and non-autonomous manners. Further analyses reveal that Talpid3 associates with Ninein to regulate microtubule organization and maintain the integrity of adherens junctions to anchor RGCs. Moreover, genetic ablation of Talpid3 results in synchronized, ectopic mitosis of neural progenitors and dysregulated neurogenesis. Our study provides an intriguing perspective for the non-ciliogenic role of centriolar proteins in mediating cortical neurogenesis.


Dichotomous role of TGF-β controls inducible regulatory T-cell fate in allergic airway disease through Smad3 and TGF-β-activated kinase 1.
Inducible CD4+CD25+ regulatory T (iTreg) cells can become pathogenic effector cells, enhancing lung allergic responses. We aimed to define the underlying cellular and molecular pathways activated by TGF-β, which determine the suppressor or enhancing activities of iTreg cells. Sensitized wild-type and CD8-deficient (CD8-/-) mice were challenged with allergen. Isolated CD4+CD25- T cells were activated by using anti-CD3/anti-CD28. To generate suppressor iTreg cells, cells were then differentiated in the presence of TGF-β, whereas IL-17-producing effector T cells were additionally exposed to IL-6. After TGF-β, Smad3 and TGF-β-activated kinase 1 (TAK1) kinase levels were monitored. The consequences of inhibiting either kinase were determined in vitro and after transfer into CD8-/- recipients. Quantitative PCR and chromatin immunoprecipitation were used to monitor gene expression and histone modifications at the retinoic acid-related orphan receptor γt (Rorγt) locus. In wild-type mice, iTreg cells suppressed lung allergic responses linked to Smad3-dependent forkhead box P3 (Foxp3) expression and IL-10 production. In the presence of IL-6, iTreg cells converted to TH17 cells, mediating a neutrophil-dependent enhancement of lung allergic responses in CD8-/- mice. Conversion was regulated by TAK1. Inhibition or silencing of TAK1 prevented expression of Rorγt and TH17 differentiation through histone modifications of Rorγt; Foxp3 expression and iTreg cell-mediated suppression remained intact. In the same cell, TGF-β induced coexpression of Smad3 and TAK1 proteins; in the presence of IL-6, expression of Smad3 and Foxp3 but not TAK1 decreased. TGF-β regulates iTreg cell outcomes through 2 distinct signal transduction pathways: one Smad3 dependent and the other TAK1 dependent. The balance of these pathways has important implications in TH17-mediated autoimmune diseases and neutrophil-dependent asthma.


Computed tomography: regression of periventricular enhancing lesions in multiple sclerosis.
A patient with two periventricular enhancing lesions on computed tomography subsequently demonstrated clearing of these abnormalities without treatment and a clinical course compatible with multiple sclerosis. CT scans obtained in disease processes associated with acute demyelination and alteration of the blood-brain barrier can show enhancing lesions. These lesions can initially be confused with primary or secondary neoplasms, enhancing infarcts or arteriovenous malformation. The differential diagnosis of enhancing lesions without significant mass effect should initially include the acute phase of multiple sclerosis and other demyelinating processes.


A Viral Protein Restricts Drosophila RNAi Immunity by Regulating Argonaute Activity and Stability.
The dicistrovirus, Cricket paralysis virus (CrPV) encodes an RNA interference (RNAi) suppressor, 1A, which modulates viral virulence. Using the Drosophila model, we combined structural, biochemical, and virological approaches to elucidate the strategies by which CrPV-1A restricts RNAi immunity. The atomic resolution structure of CrPV-1A uncovered a flexible loop that interacts with Argonaute 2 (Ago-2), thereby inhibiting Ago-2 endonuclease-dependent immunity. Mutations disrupting Ago-2 binding attenuates viral pathogenesis in wild-type but not Ago-2-deficient flies. CrPV-1A also contains a BC-box motif that enables the virus to hijack a host Cul2-Rbx1-EloBC ubiquitin ligase complex, which promotes Ago-2 degradation and virus replication. Our study uncovers a viral-based dual regulatory program that restricts antiviral immunity by direct interaction with and modulation of host proteins. While the direct inhibition of Ago-2 activity provides an efficient mechanism to establish infection, the recruitment of a ubiquitin ligase complex enables CrPV-1A to amplify Ago-2 inactivation to restrict further antiviral RNAi immunity.


Colorectal adenocarcinoma in patients less than 40 years old.
Carcinoma of the colorectum knows no age barrier. To date, more than 1,400 cases have been reported to occur in persons less than 40 years old. Our experience with 70 patients treated over a ten-year interval is reviewed. Signs and symptoms in the young are not distinctive. Diagnosis depends on a high index of suspicion and appropriate investigative procedures. Survival times are shorter in the patients who have mucinous and anaplastic tumors, and their incidences seem to be increased in this age segment. Overall survival does not significantly differ from our general experience (41 per cent, 5-year survival). Early diagnosis and prompt institution of aggressive surgical treatment can be expected to produce survival equivalent to that in patients of other ages.


Conventional grafting combined with minigrafting: a new approach.
Using a combination of 4.0-mm conventional Orentreich punch grafts and minigrafts, the author describes a methodology to achieve a very uniform, untufted look which is usually the end result of conventional grafting alone. Advantages and disadvantages are detailed.


Analysis of variants in upstream open reading frames of human globin-related genes.
β-thalassemia is an autosomal recessive monogenic disease that is caused by defects in the production of β-like globin chains. Activation of γ-globin gene and the increase in fetal hemoglobin expression have been demonstrated as one of the most important factors to ameliorate the clinical outcome of β-thalassemia patients. In this study, 202 genes or miRNAs associated with human hemoglobin gene expression from 1802 β-thalassemia patients were analyzed with target capture and next generation sequencing strategies in terms of functional variants that might affect hemoglobin gene expression. The subsequent bioinformatics analysis included assessments of sequence quality, the variants within the target regions and the 5'UTR with potential effects on upstream open reading frames (uORFs). Among the 41 variants in 5'UTR potentially affecting the uORFs identified in the study, two variants (chr19: 41859418 G > A and chr1:153606541 C > T) were experimentally validated with dual-luciferase assays to be capable of significantly down-regulating the expression of TGFB1 and CHTOP gene, respectively. The present study demonstrated a system suitable for evaluating the importance of variants in 5'UTRs affecting uORFs in 202 human genes associated with hemoglobin expression. Research with this approach could provide potential targets that may contribute to the clinical phenotypes and provide biomarkers for precise diagnosis of β-thalassemia.


Anterior chamber cleavage syndrome. A stepladder classification.
This paper classifies the abnormalities of the anterior chamber cleavage syndrome (mesodermal dysgenesis of the iris and cornea). The anatomic findings are arranged in a tabular stepladder fashion which builds from simple to more complex combinations, most of which have been previously known by eponyms. There are three groups of anomalies: 1) peripheral, 2) central, and 3) combinations of the two. 1) The peripheral anomalies consist of a prominent Schwalbe's ring, iris strands to Schwalbe's ring, and hypoplasia of the anterior iris stroma. Developmental glaucoma is commonly present. 2) The essential feature of the central anomalies is a defect in the corneal endothelium and Descemet's membrane with an overlying corneal opacity. Additional components include central iridocorneal adhesions, keratolenticular approximation with cataract, and scleralizaiton of the cornea. Chorioretinal anomalies, developmental glaucoma, and systemic malformations may be present. 3) Central and peripheral combinations may exist in the same eye, in both eyes of the same patient, or within the same family.


Left ventricular free wall rupture after coronary artery surgery.
A patient who had undergone urgent coronary artery bypass grafting for unstable angina developed a fatal left ventricular free wall rupture in the early postoperative period. Heart rupture following surgical revascularisation has not been reported before.


Risk Factors for Duty-Related Posttraumatic Stress Disorder among Police Officers in the Mt. Ontake Eruption Disaster-Support Task Force.
Mount Ontake in Nagano Prefecture, Japan erupted on 27 September 2014. Many police officers were called in for duty as a disaster-support task force. We investigated the association between the peritraumatic situation and posttraumatic stress disorder (PTSD) symptoms in these police officers. In January 2015, a health survey (OHS) on disaster stress related to the Mt. Ontake eruption disaster support work was distributed to all of the police officers and staff involved in the disaster support. We analyzed the 213 participants who had PTSD symptoms following the eruption and no missing OHS data. Logistic regression analyses were conducted to clarify the relationship between the participants' symptom severity and their peritraumatic situation (i.e., stressors and daily support prior to the eruption, disaster-support work duties, and postdisaster stress relief). The symptom severity was associated with 'more than seven cumulative days at work' (odds ratio [OR] = 2.47, 1.21-5.06), 'selecting drinking and/or smoking as stress relief after disaster-support work' (OR = 2.35, 1.09-5.04), and 'female' (OR = 3.58, 1.19-10.77). As disaster-support work, 'supporting the victims' families' (OR = 1.99, 0.95-4.21) tended to be associated with symptom severity. The number of days of disaster-support work, stress-relief behavior, and gender were associated with the severity of PTSD symptoms.


Growth inhibition and differentiation of human breast cancer cells by the PAFR antagonist WEB-2086.
WEB-2086 -- an antagonist of platelet-activating factor receptor (PAFR) with known anti-inflammatory, antiangiogenic and antileukaemic properties -- also proved to inhibit the proliferation in human solid tumour cell lines of different histology, and with much higher efficacy than in normal fibroblasts. A detailed analysis of WEB-2086 anticancer activity was then performed focusing on breast adenocarcinoma MCF-7 and MDA-MB-231 cells. WEB-2086-treated cells, either expressing (MCF-7) or unexpressing (MDA-MB-231) the oestrogen receptor (ER)alpha, underwent a dose-dependent growth arrest (IC(50)=0.65+/-0.09 and 0.41+/-0.07 mM, respectively) and accumulation in G(0)-G(1) phase. WEB-2086 also induced morphological and functional changes typical of mature mammary phenotype including (i) cell enlargement and massive neutral lipid deposition (best accomplished in MCF-7 cells); (ii) decrease in motility and active cathepsin D levels (mainly observed in highly invasive MDA-MB-231 cells). The expression of ERalpha was neither increased nor reactivated in treated MCF-7 or MDA-MB-231 cells, respectively. WEB-2086-induced differentiation in breast cancer cells involved the upregulation of PTEN, a key tumour suppressor protein opposing tumorigenesis, and was apparently independent of p53, PAFR, peripheral benzodiazepine receptor and ERalpha status. Overall, WEB-2086 can be proposed as an effective antiproliferative and differentiative agent with interesting translational opportunities to treat breast cancers in support to conventional chemotherapy.


Clioquinol promotes cancer cell toxicity through tumor necrosis factor alpha release from macrophages.
Copper has an important role in cancer growth, angiogenesis, and metastasis. Previous studies have shown that cell-permeable metal ligands, including clioquinol (CQ) and pyrrolidine dithiocarbamate, inhibit cancer cell growth in cell culture and in vivo. The mechanism of action has not been fully determined but may involve metal-mediated inhibition of cancer cell proteasome activity. However, these studies do not fully account for the ability of cell-permeable metal ligands to inhibit cancer cell growth without affecting normal cells. In this study, we examined the effect of CQ on macrophage-mediated inhibition of HeLa cancer cell growth in vitro. When CQ was added to RAW 264.7 macrophage-HeLa cell cocultures, a substantial increase in HeLa cell toxicity was observed compared with CQ treatment of HeLa cells cultured alone. Transfer of conditioned medium from CQ-treated macrophages to HeLa cells also induced HeLa cell toxicity, demonstrating the role of secreted factors in the macrophage-mediated effect. Further investigation revealed that CQ induced copper-dependent activation of macrophages and release of tumor necrosis factor (TNF) alpha. In studies with recombinant TNFalpha, we showed that the level of TNFalpha released by CQ-treated macrophages was sufficient to induce HeLa cell toxicity. Moreover, the toxic effect of conditioned medium from CQ-treated macrophages could be prevented by addition of neutralizing antibodies to TNFalpha. These studies demonstrate that CQ can induce cancer cell toxicity through metal-dependent release of TNFalpha from macrophages. Our results may help to explain the targeted inhibition of tumor growth in vivo by CQ.


[Endogenous candida endophthalmitis in a drug dependent patient: intravenous therapy with liposome encapsulated amphotericin B].
We report this exemplary case of a drug addict with candida endophthalmitis where we used the new form of amphotericin B, encapsulated in liposomes. We were able to reconfirm the reduced number of side effects and the minimized nephrotoxicity reported by authors of other specialties. In our patient, a reduction or elimination of the yeast was probably achieved, nevertheless, he developed a traction retinal detachment. In future cases of fungal endophthalmitis, we recommend liposomal amphotericin B in higher doses.


Displaced amacrine cells of the mouse retina.
The aim of this study was to characterize and classify the displaced amacrine cells in the mouse retina. Amacrine cells in the ganglion cell layer were injected with fluorescent dyes in flat-mounted retinas. Dye-filled displaced amacrine cells were classified according to dendritic field size, horizontal and vertical stratification patterns, and general morphology. We identified 10 different morphological types of displaced amacrine cell. Six of the cell types identified here are novel cell types that have not been described previously in the mouse retina, to the best of our knowledge. The displaced amacrine cells included four types of medium-field cells, with dendritic field diameters of 200-500 microm, and six types of wide-field cells, with dendritic fields extending over 500 microm. Narrow-field displaced amacrine cells, with dendritic field diameters smaller than 200 microm, were not encountered. The most frequently labeled displaced amacrine cell type was the starburst amacrine cell. At least three cell types identified here have nondisplaced counterparts in the inner nuclear layer as well. Displaced amacrine cells display a rich variety of stratification and branching patterns, which surely reflect the wide range of their functional roles in the processing of visual signals in the inner retina.


Antiproliferative activity of olomoucine II, a novel 2,6,9-trisubstituted purine cyclin-dependent kinase inhibitor.
The study describes the protein kinase selectivity profile, as well as the binding mode of olomoucine II in the catalytic cleft of CDK2, as determined from cocrystal analysis. Apart from the main cell cycle-regulating kinase CDK2, olomoucine II exerts specificity for CDK7 and CDK9, with important functions in the regulation of RNA transcription. In vitro anticancer activity of the inhibitor in a panel of tumor cell lines shows a wide potency range with a slight preference for cells harboring a wild-type p53 gene. Cell-based assays confirmed activation of p53 protein levels and events leading to accumulation of p21(WAF1). Additionally, in olomoucine II-treated cells, Mdm2 was found to form a complex with the ribosomal protein L11, which inhibits Mdm2 ubiquitin ligase function. We conclude that perturbations in RNA synthesis may lead to activation of p53 and that this contributes to the antiproliferative potency of cyclindependent kinase inhibitors.


A randomized trial of topical mitomycin-C in the immediate post-operative period for the short and long-term prevention of granulation tissue in mastoid cavities.
Granulation tissue is a primary source of otorrhea in the ear, particularly after surgery, and may develop complications in patients. Hence, discovering an efficient treatment is crucial for this disorder. This study aimed to evaluate the mitomycin drops efficacy for germination prevention during the recovery process after mastoidectomy. Our purpose was to introduce novel treatment options in some conditions in which we could not reconstitute the cavity to prevent the possible germination. This clinical trial was performed on 52 patients undergoing mastoidectomy. Patients were randomly divided into two groups (26 patients in the mitomycin group and 26 patients in the placebo group). After drug administration, the granulation tissue and complications were recorded. All statistical analysis was performed using SPSS version 21. The results of our study showed that mitomycin administration in patients undergoing mastoidectomy significantly reduced the formation of the granulation tissue in the first and third months after surgery (P < 0.05), which is associated with increased patient satisfaction (P < 0.05). The germ formation after mastoidectomy is so common. Since mitomycin administration decreased the incidence of germ formation, it can be proposed as a suggestive treatment in all patients to increase surgical quality and decrease complications include granulation formation.


Disease progression in melanoma patients with negative sentinel lymph node: does false-negative specimens entirely account for this phenomenon?
Sentinel lymph node (SLN) status is the most important prognostic factor for subjects with primary melanoma thicker than 1 mm. We focused our study on patients with disease progression after negative SLN biopsy (SLNB), with the aim of elucidating their clinical and histopathological characteristics, outcome and real incidence of false negative. A total of 688 melanoma patients who underwent SLNB (1 May 1998-31 December 2008) were analysed; all patients had Breslow >1 mm or Breslow <1 mm and at least one of the following features: regression, ulceration and/or Clark level IV-V. Progression developed in 114 of 503 negative SLN patients (22.7%); the first metastatic site was regional in 64% and distant in 36% of these cases. Thirty-nine patients had nodal metastases in the SLN basin as first site of progression. High-risk melanomas (P = 0.001) and elderly patients (P = 0.0005) had an increased probability of progression. Women with a higher median age and lower limbs primary melanoma developed mainly regional skin metastases, while an increased probability of distant metastases was demonstrated in patients with primary on the trunk and axillary SLN (P = 0.003, P = 0.001 respectively). Age at diagnosis, Breslow thickness and regression showed a prognostic relevance in univariate and multivariate analyses on disease-free survival and overall survival. Even if SLN status remains the most important prognostic factor for melanoma patients, progressive disease after a negative SLNB is a relatively frequent event. However, in our opinion, only a part of negative SLNB patients with metastatic spreading should be considered as false negative (7.75%).


Hair analysis for drugs of abuse. V. The facility in incorporation of cocaine into hair over its major metabolites, benzoylecgonine and ecgonine methyl ester.
We studied the incorporation of cocaine (COC), benzoylecgonine (BE) and ecgonine methyl ester (EME) into hair from blood. At first, the time courses of three drugs in rat plasma following i.p. administration of cocaine were investigated over 360 min. AUCs of COC, BE and EME in plasma were 14.2, 60.7 and 53.4 micrograms/ml/min, respectively. In contrast, the concentrations of the three compounds in hair were 16.4, 1.7 and 0.8 ng/mg. In spite of that, the AUC of COC in plasma was much lower than that of the other two compounds in plasma, the concentration of COC in hair was much higher than that of the other two compounds. The incorporation of COC into hair was much greater than that of BE and EME. If the incorporation of drugs from blood into hair is compared by [concentration in hair]/[AUC in plasma], that of COC, BE and EME is represented by 77:1.9:1. Our results suggest that the incorporation of drugs into hair from blood unexpectedly depends upon the physical properties of each drug.


Limited versus Extended Pelvic Lymph Node Dissection for Prostate Cancer: A Randomized Clinical Trial.
Pelvic lymph node dissection (PLND) is the most reliable procedure for lymph node staging. However, the therapeutic benefit remains unproven; although most radical prostatectomies at academic centers are accompanied by PLND, there is no consensus regarding the optimal anatomical extent of PLND. To evaluate whether extended PLND results in a lower biochemical recurrence rate. We conducted a single-center randomized trial. Patients, enrolled between October 2011 and March 2017, were scheduled to undergo radical prostatectomy and PLND. Patients were assigned to limited or extended PLND by cluster randomization. Specifically, surgeons were randomized to perform limited or extended PLND for 3-mo periods. Randomization to limited (external iliac nodes) or extended (external iliac, obturator fossa and hypogastric nodes) PLND. The primary endpoint was the rate of biochemical recurrence. Of 1440 patients included in the final analysis, 700 were randomized to limited PLND and 740 to extended PLND. The median number of nodes retrieved was 12 (interquartile range [IQR] 8-17) for limited PLND and 14 (IQR 10-20) extended PLND; the corresponding rate of positive nodes was 12% and 14% (difference -1.9%, 95% confidence interval [CI] -5.4% to 1.5%; p = 0.3). With median follow-up of 3.1 yr, there was no significant difference in the rate of biochemical recurrence between the groups (hazard ratio 1.04, 95% CI 0.93-1.15; p = 0.5). Rates for grade 2 and 3 complications were similar at 7.3% for limited versus 6.4% for extended PLND; there were no grade 4 or 5 complications. Extended PLND did not improve freedom from biochemical recurrence over limited PLND for men with clinically localized prostate cancer. However, there were smaller than expected differences in nodal count and the rate of positive nodes between the two templates. A randomized trial comparing PLND to no node dissection is warranted. In this clinical trial we did not find a difference in the rate of biochemical recurrence of prostate cancer between limited and extended dissection of lymph nodes in the pelvis. This study is registered on ClinicalTrials.gov as NCT01407263.


Crucial role of vinexin for keratinocyte migration in vitro and epidermal wound healing in vivo.
In the process of tissue injury and repair, epithelial cells rapidly migrate and form epithelial sheets. Vinexin is a cytoplasmic molecule of the integrin-containing cell adhesion complex localized at focal contacts in vitro. Here, we investigated the roles of vinexin in keratinocyte migration in vitro and wound healing in vivo. Vinexin knockdown using siRNA delayed migration of both HaCaT human keratinocytes and A431 epidermoid carcinoma cells in scratch assay but did not affect cell proliferation. Induction of cell migration by scratching the confluent monolayer culture of these cells activated both EGFR and ERK, and their inhibitors AG1478 and U0126 substantially suppressed scratch-induced keratinocyte migration. Vinexin knockdown in these cells inhibited the scratch-induced activation of EGFR, but not that of ERK, suggesting that vinexin promotes cell migration via activation of EGFR. We further generated vinexin (-/-) mice and isolated their keratinocytes. They similarly showed slow migration in scratch assay. Furthermore, vinexin (-/-) mice exhibited a delay in cutaneous wound healing in both the back skin and tail without affecting the proliferation of keratinocytes. Together, these results strongly suggest a crucial role of vinexin in keratinocyte migration in vitro and cutaneous wound healing in vivo.


Clearing a path through the patent thicket.
Patents do not always promote innovation, particularly when they restrict access to fundamental scientific discoveries and the tools of basic research. However, there are legal and policy approaches that may help to ameliorate problems associated with patenting these sorts of inventions.


Ultrasound therapy for treatment of lower extremity intermittent claudication.
While often thought of as a diagnostic tool, ultrasound (US) can also potentially be used as a therapeutic modality. US applies mechanical stress on endothelial cells and induces nitric oxide synthase, which regulates the secretion of nitric oxide, a potent vasodilator. In animal ischemic models, US has been shown to improve hindlimb, myocardial, and cerebral perfusion. We performed a pilot trial of US therapy in the lower extremities of human subjects with intermittent claudication. 10 subjects (5 male, 5 female, mean age 69.7 ± 10.3) with intermittent claudication were recruited. Both legs were placed in a specially designed boot with a water interface between US transducers and the legs. Subjects underwent pulsed US therapy at 250 kHz frequency for 30 min for three treatments a week for six weeks. Pre and post treatment ankle:brachial index (ABI), 6-min walk (6 MW), Walking Impairment Questionnaire (WIQ), and Short Form 36 (SF36) were performed. Pre and post-treatment results were compared with paired t-test. Six minute walking distance at baseline was 352 ± 70 m, after one treatment session 353 ± 70 m (p = 0.99), and at completion 372 ± 71 m (p = 0.015). There was a trend toward improved ABI after 6 weeks of treatment (0.53 ± 0.17 vs 0.64 ± 0.12, p = 0.083). After six weeks, significant improvements were noted in overall WIQ score (2.00 ± 1.48 vs 2.63 ± 1.38, p = 0.0001), WIQ (distance) 2.07 ± 1.54 vs 2.73 ± 1.42 (p = 0.036), and WIQ (stair) 2.00 ± 1.67 vs 2.62 ± 1.24, p = 0.034, with a trend in WIQ (speed), 1.89 ± 1.26 vs 2.46 ± 1.43, p = 0.069. In the SF-36, significant improvements were noted in the domains of physical functioning (44.0 ± 41.6 vs 50.5 ± 41.1, p = 0.009) and role limitations - physical (35.0 ± 48.3 vs 60.0 ± 49.6, p = 0.006) after six weeks. Therapeutic US is a potential noninvasive treatment for intermittent claudication. Pilot study patients noted significant improvements in 6 MW and WIQ results after 6 weeks of treatment. A nonsignificant improvement in ABI was noted. Further research will be needed to clarify optimal treatment frequency and duration.


Introducing a nursing maintenance bundle for patients with pulmonary arterial catheters.
We undertook a quality improvement project to address challenges with pulmonary artery catheter (PAC) line maintenance in a setting of low-baseline central-line infection rates. We observed a subsequent reduction in Staphylococcal PAC line infections and a trend toward a reduction in overall PAC infection rates over 1 year.


[Monoclonal human immunoglobulin (IgG lambda) with antiethinylestradiol activity, oral contraceptives, and arterial pulmonary thrombosis].
In a 36-year-old woman taking an oral contraceptive containing 50 mug of ethinyloestradiol each day, a pulmonary arterial thrombosis and a monoclonal gammapathia were associated. The monoclonal IgI lambda Mai... was prepared. When purified, this IgG lambda binds ethinyloestradiol with strong affinity (Ka= 2.7 times 10(7)M-1) and also 17-beta-oestradiol with a little less affinity (Ka = 0.4 times 10(7)M-1. For those ligands each IgG lambda Mai... molecule has two sites of same affinity and specificity so that a Scatchard plot of the experimental values gives a straight line. It is likely that the antibody sites of the IgG lambda Mai... are the binding sites. These facts support the hypothesis of an immunological mechanism of the thromboembolic disease which may be induced by oral contraceptives.


Physiological investigations of the influences of byproduct pathways on 3-hydroxypropionic acid production in Klebsiella pneumoniae.
Klebsiella pneumoniae can naturally synthesize 3-hydroxypropionic acid (3-HP), 1,3-propanediol (1,3-PD), and 2,3-butanediol (2,3-BD) from glycerol. However, biosynthesis of these industrially important chemicals is constrained by troublesome byproducts. To clarify the influences of byproducts on 3-HP production, in this study, a total of eight byproduct-producing enzyme genes including pmd, poxB, frdB, fumC, dhaT, ilvH, adhP, and pflB were individually deleted from the K. pneumoniae genome. The resultant eight mutants presented different levels of metabolites. In 24-h shake-flask cultivation, the adhP- and pflB-deletion mutants produced 0.41 and 0.44 g/L 3-HP, respectively. Notably, the adhP and pflB double deletion mutant K. pneumoniaeΔadhPΔpflB produced 1.58 g/L 3-HP in 24-h shake-flask cultivation. When K. pneumoniaeΔadhPΔpflB was harnessed as a host strain to overexpress PuuC, a native aldehyde dehydrogenase (ALDH) catalyzing 3-hydroxypropionaldehyde (3-HPA) to 3-HP, the resulting recombinant strain K. pneumoniaeΔadhPΔpflB(pTAC-puuC) (pTAC-puuC is PuuC expression vector) generated 66.91 g/L 3-HP with a cumulative yield of 70.84% on glycerol in 60-h bioreactor cultivation. Additionally, this strain showed 2.3-, 5.1-, and 0.67-fold decrease in the concentrations of 1,3-PD, 2,3-BD, and acetic acid compared with the reference strain K. pneumoniae(pTAC-puuC). These results indicated that the byproducts exerted differential impacts on the production of 3-HP, 1,3-PD, and 2,3-BD. Although combinatorial elimination of byproduct pathways could reprogram glycerol flux, the enzyme 1,3-propanediol oxidoreductase (DhaT) that catalyzes 3-HPA to 1,3-PD and the enzymes ALDHs, especially, PuuC are most pivotal for 3-HP production. This study provides a deep understanding of how byproducts affect the production of 3-HP, 1,3-PD, and 2,3-BD in K. pneumoniae.


The prognostic value of biomarkers after a premature myocardial infarction.
Intravascular thrombogenesis is influenced by a complex interplay of factors related to a procoagulant state, fibrinolysis, endothelial damage/dysfunction and inflammation. We hypothesised that abnormalities of these biological systems would contribute to outcome of coronary artery disease presenting at a young age. We performed a prospective study of 142 subjects presenting with acute myocardial infarction (AMI) at a young age (defined as age ≤45 years), to determine the clinical and laboratory predictors of cardiovascular events during 36 months of follow-up. We assessed conventional risk factors and abnormalities of thrombophilia [total homocysteine (tHcy), lipoprotein (a) [Lp(a)], antiphospholipid antibodies (APA)], as well as lipid profile (total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides), fibrinogen and fibrin D-dimer (as indices of a hypercoagulable state and thrombogenesis), von Willebrand factor (vWF, an index of endothelial damage/dysfunction), tissue plasminogen activator [t-PA antigen] and its inhibitor [PAI-1 antigen] (as indices of fibrinolysis), and C-reactive protein [CRP] (an index of inflammation). In a multivariate analysis, the variables independently associated with cardiovascular events at follow-up were levels of homocysteine (OR 3.73, 95% CI (1.54-9.02); p=0.003), left ventricle systolic dysfunction (OR 3.04, 95% CI (1.00-9.25); p=0.050), and smoking habit (OR 2.79, 95% CI (1.09-7.14) p=0.032). Prognostic markers associated with cardiovascular events in premature CAD (young AMI subjects) were cigarette smoking and EF<50% of left ventricle as conventional clinical risk factors, as well as higher levels of homocysteine.


Coordinated AR and microRNA regulation in prostate cancer.
The androgen receptor (AR) remains a key driver of prostate cancer (PCa) progression, even in the advanced castrate-resistant stage, where testicular androgens are absent. It is therefore of critical importance to understand the molecular mechanisms governing its activity and regulation during prostate tumourigenesis. MicroRNAs (miRs) are small ∼22 nt non-coding RNAs that regulate target gene, often through association with 3' untranslated regions (3'UTRs) of transcripts. They display dysregulation during cancer progression, can function as oncogenes or tumour suppressors, and are increasingly recognised as targets or regulators of hormonal action. Thus, understanding factors which modulate miRs synthesis is essential. There is increasing evidence for complex and dynamic bi-directional cross-talk between the multi-step miR biogenesis cascade and the AR signalling axis in PCa. This review summarises the wealth of mechanisms by which miRs are regulated by AR, and conversely, how miRs impact AR's transcriptional activity, including that of AR splice variants. In addition, we assess the implications of the convergence of these pathways on the clinical employment of miRs as PCa biomarkers and therapeutic targets.


Impact of urban morphology on the spatial and temporal distribution of PM2.5 concentration: A numerical simulation with WRF/CMAQ model in Wuhan, China.
The urban morphology can significantly change the urban microclimate, which in turn affects the diffusion of air pollutants. Urban planning is the most important means of shaping urban morphology. Therefore, this study takes Wuhan as an example and uses the method of WRF/CMAQ coupled UCM model to analyze the spatial and temporal distribution characteristics of PM2.5 in the Wuhan metropolitan area in winter 2015. The six most important urban morphological indicators in urban planning: the floor area ratio and building height, building density and building width, vegetation coverage ratio, and urban fraction, are selected and classified into three groups. Studying their impact on the spatial and temporal distribution of PM2.5 concentration provides support for urban planners to improve air quality. The results show that the maximum value of PM2.5 concentration in Wuhan urban area occurs in the morning rush hour, and PM2.5 is distributed concentrically in the downtown of the city (within the second ring highway) according to the highways around the city. The PM2.5 concentration in the downtown area with the most extensive urban morphological index is the highest, and it decreases with increasing distance from the downtown. Among the six indicators, building density and urban fraction have the most significant impact on PM2.5 concentration because they have the greatest impact on the wind speed at 10 m. The height of the planetary boundary layer is the key factors affect the vertical and horizontal diffusion of air pollutants. Except for the vegetation coverage ratio, the increase of other urban morphological indicators will lead to a decrease of PM2.5 concentration in Wuhan urban area at night. During the daytime, increasing the floor area ratio and building height will cause an increasing of PM2.5 concentration, but other indicators have the opposite effects.


Staphylococcus aureus Bacteremia Risk in Hemodialysis Patients Using the Buttonhole Cannulation Technique: A Prospective Multicenter Study.
Arteriovenous fistulas are the preferred access type for hemodialysis. The buttonhole needling technique has become an alternative to stepladder or area puncture. However, an increased risk for infection has been described. The present study examined the risk for infectious complications with different needling techniques. Prospective multicenter observational cohort study with 5 years of follow-up. In-center hemodialysis patients from 5 hemodialysis units in Denmark, dialyzed on a native arteriovenous fistula. 286 patients were included; 144 cannulated with the buttonhole technique. The buttonhole cannulation technique was compared to the stepladder or area puncture technique. Primary end points: event rates of access-related Staphylococcus aureus bacteremia and the HR for first access-related S aureus bacteremia. Secondary end points: local infections and access-related S aureus bacteremia-related metastatic infections and mortality. Time-to-event analysis using Cox proportional hazards regression to estimate the HR of access-related S aureus bacteremia in buttonhole cannulation compared to stepladder/area puncture. Poisson regression was used for incidence rate ratio calculations. S aureus caused 48 access-related bacteremias; 43 (90%) in the buttonhole group compared with 5 (10%) in the stepladder/area group. The HR for first access-related S aureus bacteremia was significantly higher for buttonhole cannulation compared to stepladder/area needling (unadjusted, 6.8 [95% CI, 2.4-19.1]; adjusted, 8.4 [95% CI, 2.9-24.2]). The incidence rate ratio for access-related S aureus bacteremia was 6.8 (95% CI, 2.9-16.1), and the incidence rate ratio of local cannulation-site infection without access-related S aureus bacteremia was 3.8 (95% CI, 1.3-15.4) for buttonhole cannulation compared to stepladder/area needling. Nonrandomized observational design, prevalent hemodialysis patients. Access-related S aureus bacteremia rates were very high for buttonhole cannulation compared to stepladder/area needling, questioning the use of buttonhole cannulation in routine clinical practice. A restrictive approach to buttonhole use is recommended, with buttonhole cannulation only being used as a second alternative to area technique when stepladder cannulation is not feasible.


Emergency Physicians' Workstation Design: An Observational Study of Interruptions and Perception of Collaboration During Shift-End Handoffs.
Frequent external interruptions and lack of collaboration among team members are known to be common barriers in end-of-shift handoffs between physicians in the emergency department. In spite of being the primary location for this crucial and cognitively demanding task, workstations are not designed to limit barriers and support handoffs. The purpose of this study is to examine handoff characteristics, actual and perceived interruptions, and perceived collaboration among emergency physicians performing end-of-shift handoffs in physician workstations with varying levels of enclosures-(a) open-plan workstation, (b) enclosed workstation, and (c) semi-open workstation. Handoff and interruption characteristics were captured through in-person observations of 60 handoffs using an iPad-based tool. Additionally, physicians participating in the handoffs responded to a survey pertaining to their perception of interruptions and collaboration with clinicians during each phase. Other organizational and demographic data were obtained from the hospital database, surveys, and observations. Physicians working in the open workstation experienced a significantly higher number of interruptions/hour (18.08 int/hr) as compared to the semi-open (13.62 int/hr) and enclosed workstations (11.41 int/hr). Most physicians perceived that they were interrupted in the semi-open and open workstations. In addition, majority of physicians in the enclosed pod perceived high collaboration with clinicians involved in and present in the workstation during handoff. This correlational study showed positive outcomes experienced by physician working in the enclosed workstation as compared to the open and semi-open workstations.


Outsourcing a visual neuropil - The central visual system of the median eyes of Galeodes granti Pocock, 1903 (Arachnida: Solifugae).
Only a few studies have examined the central visual system of Solifugae until now. To get new insights suitable for phylogenetic analysis we studied the R-cell (or retinula cell) projections and visual neuropils of Galeodes granti using various methods. G. granti possesses large median eyes and rudimentary lateral eyes. In this study, only the R-cells and neuropils of the median eyes were successfully stained. The R-cells terminate in two distinct visual neuropils. The first neuropil is located externally to the protocerebrum directly below the retina, the second neuropil lies in the cell body rind of the protocerebrum, and immediately adjacent is the arcuate body. This layout of the median eye visual system differs from Arachnopulmonata (Scorpiones + Tetrapulmonata). However, there are several similarities with Opiliones. In both, (1) the R-cells are connected to a first and second visual neuropil and not to any other region of the brain, (2) the first neuropil is not embedded in the cell body rind of the protocerebrum, it is rather external to the protocerebrum, (3) the second visual neuropil is embedded in the cell body rind, and (4) the second neuropil abuts the arcuate body. These findings may provide important new characters for the discussion on arachnid phylogeny.


A nurse goes shopping for a continuing care retirement community.
The selection of a continuing care retirement community (CCRC) in which to live is not a matter of flipping a coin. It involves serious and time-consuming investigation. For most people, it is a daunting task. Elderly people who decide on a CCRC for their living arrangement need to follow a selection process to arrive at the appropriate choice--the choice that best meets their particular needs and provides quality of life. This personal account gives insight into one person's experience with the process. Nurses who are knowledgeable about the types of CCRCs available are in a position to help their patients make more informed choices.


Structural and dynamic properties of bacterial type IV secretion systems (review).
The type IV secretion systems (T4SS) are widely distributed among the gram-negative and -positive bacteria. These systems mediate the transfer of DNA and protein substrates across the cell envelope to bacterial or eukaryotic cells generally through a process requiring direct cell-to-cell contact. Bacteria have evolved T4SS for survival during establishment of pathogenic or symbiotic relationships with eukaryotic hosts. The Agrobacterium tumefaciens VirB/D4 T4SS and related conjugation machines serve as models for detailed mechanistic studies aimed at elucidating the nature of translocation signals, machine assembly pathways and architectures, and the dynamics of substrate translocation. The A. tumefaciens VirB/D4 T4SS are polar-localized organelles composed of a secretion channel and an extracellular T pilus. These T4SS are assembled from 11 or more subunits. whose membrane topologies, intersubunit contacts and, in some cases, 3-dimensional structures are known. Recently, powerful in vivo assays have identified C-terminal translocation signals, defined for the first time the translocation route for a DNA substrate through a type IV secretion channel, and supplied evidence that ATP energy consumption contributes to a late stage of machine morphogenesis. Together, these recent findings describe the mechanics of type IV secretion in unprecedented detail.


Optimizing the imaging of the monkey auditory cortex: sparse vs. continuous fMRI.
The noninvasive imaging of the monkey auditory system with functional magnetic resonance imaging (fMRI) can bridge the gap between electrophysiological studies in monkeys and imaging studies in humans. Some of the recent imaging of monkey auditory cortical and subcortical structures relies on a technique of "sparse imaging," which was developed in human studies to sidestep the negative influence of scanner noise by adding periods of silence in between volume acquisition. Among the various aspects that have gone into the ongoing optimization of fMRI of the monkey auditory cortex, replacing the more common continuous-imaging paradigm with sparse imaging seemed to us to make the most obvious difference in the amount of activity that we could reliably obtain from awake or anesthetized animals. Here, we directly compare the sparse- and continuous-imaging paradigms in anesthetized animals. We document a strikingly greater auditory response with sparse imaging, both quantitatively and qualitatively, which includes a more expansive and robust tonotopic organization. There were instances where continuous imaging could better reveal organizational properties that sparse imaging missed, such as aspects of the hierarchical organization of auditory cortex. We consider the choice of imaging paradigm as a key component in optimizing the fMRI of the monkey auditory cortex.


Metabolic heterogeneous zone assessed by 18 FDG-PET is predictive of postablation mortality in patients with ventricular tachycardia.
We sought to study the predictive value of the metabolic heterogeneous zone (HZ) as determined by 18 Fluorodeoxyglucose (18 FDG) positron emission tomography (PET) viability studies in ventricular tachycardia (VT) patients. PET studies utilizing 82 Rubidium (82 Rb) tracer for perfusion and 18 FDG tracer for viability were analyzed using PMOD (PMOD Technologies) and further analyzed using 684-segment plots. 18 FDG uptake was normalized to the area with maximal perfusion on the rest 82 Rb study. Metabolic scar, HZ, and healthy segments were defined with perfusion-normalized 18 FDG uptake between 0%-50%, 50%-70%, and >70%, respectively. Thirty-four VT patients (age, 63 ± 12 years) were evaluated with 18 FDG-PET viability study. Most (n = 31) patients underwent VT ablation. Patients were categorized to HZ < median versus HZ ≥ median based on a median HZ area size of 21.0 cm2 . HZ size was significantly larger in the deceased group than the alive group (35.2 cm2 vs. 18.1 cm2 , p = .01). Deaths were significantly higher in HZ ≥ 21 cm2 group than HZ < 21 cm2 group (58.8% vs. 11.8%, p = .005). Survival analysis showed significantly higher mortality in the HZ ≥ 21 cm2 group than the HZ < 21 cm2 group (HR = 4.1, 95% CI: 1.3-12.6, p = .016). In a multivariable analysis, HZ was found to be an independent predictor for all-cause mortality (HR = 1.07, 95% CI: 1.02-1.12, p = .01) CONCLUSIONS: Increased HZ size of myocardium was associated with increased mortality. Metabolic HZ quantification may be of value in risk stratification and management of ischemic and nonischemic patients with VT.


Free calcium wave upon activation in Xenopus eggs.
Eggs of Xenopus laevis were preloaded with aequorin and the spatial and temporal pattern of free calcium release in the egg cortex on artificial activation was determined by the aequorin luminescence emitted from the thin cortical layer of naturally opaque eggs. The aequorin luminescence was detected with a photonic microscope system consisting of a light microscope and a two-dimensional photon-counting system with an image processor. A free calcium increase was initiated around the point of prick activation. The state of increased Ca2+ propagated in the cortical cytoplasm of the egg as a wave with a velocity of about 8 micron/sec at 22 degrees C. This wave reached the antipode by 5 to 6 min of prick activation. The spatial pattern of the Ca2+ wave was similar to that of changes in brightness of the egg surface on activation, termed the "activation wave" by K. Hara and P. Tydeman (1979, Wilhelm Roux's Arch. Dev. Biol. 186, 91-94). To examine the temporal correlation between the Ca2+ wave and the activation wave, images of aequorin luminescence and those of the egg cortex taken by incident light illumination were recorded alternately in the same egg. The zone of free calcium increase corresponded to the light (relaxation) zone of the activation wave, where exocytosis of cortical granules and elongation of microvilli were taking place.


Methyltrioxorhenium-catalyzed epoxidation of homoallylic alcohols with hydrogen peroxide.
Homoallylic alcohols were efficiently converted to the corresponding 3,4-epoxy alcohols in excellent yields by methyltrioxorhenium (MTO)-catalyzed epoxidation with aqueous hydrogen peroxide as the terminal oxidant and 3-methylpyrazole (10 mol %) as an additive. The epoxidations of homoallylic alcohols proceeded under organic solvent-free conditions faster than those in dichloromethane.


MiREDiBase, a manually curated database of validated and putative editing events in microRNAs.
MicroRNAs (miRNAs) are regulatory small non-coding RNAs that function as translational repressors. MiRNAs are involved in most cellular processes, and their expression and function are presided by several factors. Amongst, miRNA editing is an epitranscriptional modification that alters the original nucleotide sequence of selected miRNAs, possibly influencing their biogenesis and target-binding ability. A-to-I and C-to-U RNA editing are recognized as the canonical types, with the A-to-I type being the predominant one. Albeit some bioinformatics resources have been implemented to collect RNA editing data, it still lacks a comprehensive resource explicitly dedicated to miRNA editing. Here, we present MiREDiBase, a manually curated catalog of editing events in miRNAs. The current version includes 3,059 unique validated and putative editing sites from 626 pre-miRNAs in humans and three primates. Editing events in mature human miRNAs are supplied with miRNA-target predictions and enrichment analysis, while minimum free energy structures are inferred for edited pre-miRNAs. MiREDiBase represents a valuable tool for cell biology and biomedical research and will be continuously updated and expanded at https://ncrnaome.osumc.edu/miredibase .


Aetna's California Case Puts New Focus on Prior Authorization.
The CNN story once again puts controversy about prior authorization into the spotlight. But is it necessary to review a patient's entire medical record?


Variation in Polycyclic Aromatic Hydrocarbon Contamination Between Native and Introduced Species of Fishes of Pallikaranai Wetland, Chennai, Tamil Nadu, India.
The impacts of invasive fish species on recipient communities are mostly considered a threat to the biodiversity of freshwater systems. A characteristic of introduced species turning invasive is their higher tolerance to environmental stressors. To understand if non-native fishes in Pallikaranai wetland, Tamil Nadu, India, are more tolerant to anthropogenic pressures in the Wetland, we assessed the variation in Polycyclic Aromatic Hydrocarbon contamination between introduced and native fish species. Mean levels of Naphthalene, fluorene, phenanthrene, anthracene, pyrene, and total PAH in samples of introduced species were higher than native species, while High Molecular weight PAH and carcinogenic PAH4 were higher in native fish species. The data was also analysed to assess if co-variates (organs, seasons, and sex) affected PAH accumulation patterns in non-native species. It was observed that only organs, sex, and year contributed significantly.


Breastfeeding and the risk of life-threatening enterotoxigenic Escherichia coli diarrhea in Bangladeshi infants and children.
To assess the relationship between breastfeeding and the risk of life-threatening enterotoxigenic Escherichia coli (ETEC) diarrhea among Bangladeshi infants and young children <36 months of age. Case-control study. A rural Bangladesh community. A total of 168 cases with clinically severe ETEC diarrhea detected in a treatment center-based surveillance system during 1985 to 1986 and 3679 controls selected in three surveys of the same community during the same calendar interval. Cases and controls were compared for the frequency of antecedent breastfeeding patterns. Compared with other feeding modes, exclusive breastfeeding of infants was associated with significant protection against severe ETEC diarrhea (relative risk [RR] = 0.51; 95% confidence interval [CI]: 0.28,0.96). However, during the second and third years of life, the risk of this outcome was similar in both breastfed and nonbreastfed children (RR = 0.98; 95% CI: 0.45,2.12), and no significant overall protective association between breastfeeding and severe ETEC diarrhea was evident during the first 3 years of life (RR = 0.86; 95% CI: 0.43,1. 74). Exclusive breastfeeding appeared to protect infants against severe ETEC diarrhea, but breastfeeding was not associated with protection after infancy, nor was it associated with a major overall reduction of severe ETEC disease during the first 3 years of life. Although not diminishing the importance of breastfeeding, our findings suggest that other interventions, such as immunization and education about proper food hygiene, may also be required in efforts to prevent this major pediatric disease.


Should the upper tracts be imaged for microscopic haematuria?
To determine the relative prevalence of various definitions of microscopic haematuria (MH) in patients with renal neoplasms and controls, and to predict the likely outcome of renal imaging for those definitions. In a retrospective case-control study 278 adult men and woman seen between 1998 and 2003 with untreated renal neoplasms were compared to controls matched for age and sex. All cases and controls had renal imaging within 6 months of a urine analysis. Patients were excluded for gross haematuria or other conditions associated with MH but not relevant to upper tract imaging. Adjusted odds ratios (OR) computed for 13 definitions of MH by conditional logistic regression were the primary outcome measures. Additional outcome measures were ORs in selected subsets. Hypothetical performance characteristics of a positive urine analysis were then derived to predict the likely results of detecting renal neoplasms for each definition of MH. The OR (95% confidence interval) for the entire series of cases and controls, both symptomatic and asymptomatic, was 2.0 (1.02-3.92, P = 0.04) for MH defined as > or = 4 red blood cells per high-power field (RBC/HPF) and 2.2 (1.09-4.52, P = 0.03) for > or = 5 RBC/HPF. No significant OR was calculated for < or = 3 RBC/HPF, nor for a subgroup of patients with MH in a routine urine analysis obtained during a periodic health examination. Symptomatic patients had an OR of 13.68 (1.6-117.1, P = 0.02) for MH defined as > or = 5 RBC/HPF. The sensitivity of a positive test decreased from 24.8% to 5.04% as the definition for MH became more stringent. The theoretical positive predictive value (assuming a prevalence of renal cell neoplasms of 0.25%) of the most stringent definition of MH was 0.58%. Patients with renal neoplasms have about twice the prevalence of MH with > or = 4 or 5 RBC/HPF in a single urine sample compared with matched controls, but this difference has little impact on the hypothetical detection rate of renal cancer. Imaging the kidney for low-grade MH in a routine urine analysis discovered at a periodic health examination in an otherwise asymptomatic patient is tantamount to screening without cause, and can be deferred for selected patients. The clinical context is as important as the degree of MH when deciding to image the kidneys.


A vaccine against methamphetamine attenuates its behavioral effects in mice.
Vaccines have treatment potential for methamphetamine (MA) addiction. We tested whether a conjugate vaccine against MA (succinyl-methamphetamine-keyhole limpet hemocyanin carrier protein; SMA-KLH) would generate MA antibodies and alter MA-induced behaviors. Mice were injected with SMA-KLH and received booster administrations 3 and 20 weeks later. Serum antibody titers reached peak levels by 4-6 weeks, remained at a modest level through 18 weeks, peaked again at 22 weeks after the second boost, and were still elevated at 35 weeks. At 7 weeks, groups of vaccinated and non-vaccinated mice were administered one of three MA doses (1, 2 or 3 mg/kg) to assess locomotor activity. Non-vaccinated mice showed dose-dependent effects of MA with hypolocomotion at the lowest dose and elevated activity levels at the highest dose. Both dose effects were reduced in SMA-KLH groups, particularly low dose-induced hypolocomotion at later times post MA administration. Separate groups of vaccinated and non-vaccinated mice were trained in MA place conditioning at 30 weeks with either 0 (vehicle) or 0.5mg/kg MA. Although times spent in the MA-paired side did not differ between groups on test vs. baseline sessions, SMA-KLH mice conditioned with MA showed reduced conditioned approach behaviors and decreased conditioned activity levels compared to control groups. These data suggest SMA-KLH attenuates the ability of MA to support place conditioning and reduces or delays its locomotor effects. Overall, results support SMA-KLH as a candidate MA vaccine.


Early-Phase Interventional Trials in Oral Cancer Prevention.
The increasing breadth of molecular targets, promise of immune-targeted therapies and repurposed agents have heightened interest in cancer prevention. While, to date, testing of oral cancer chemoprevention strategies has failed to deliver therapeutic agents for routine clinical practice, there remains an urgent need for further clinical research to overcome this hurdle. Patients at the greatest risk of disease stand to benefit the most from inclusion in clinical trials; therefore, there is a need to carefully define this population using validated clinical and molecular markers. Safety, tolerability and the efficacy of interventions is assessed through carefully selected endpoints. These endpoints may include pharmacodynamic, clinical, histological and on-target molecular modifications as an individual or as a composite endpoint. Early-phase trials provide an area of opportunity to explore novel and repurposed agents in the setting of oral cancer chemoprevention, eventually leading to phase III trials with clinical endpoints such as transformation and clinical outcome; these studies are large, lengthy and expensive and should be reserved for the most promising of agents. This paper will explore current evidence in oral cancer chemoprevention, drug repurposing, selection of appropriate endpoints for early-phase trials and novel therapeutic angles in oral cancer chemoprevention.


Risk factors for intrahepatic and extrahepatic cholangiocarcinoma: A systematic review and meta-analysis.
Cholangiocarcinoma (CCA) carries a poor prognosis, is increasing in incidence and its causes are poorly understood. Although some risk factors are known, they vary globally and collectively account for a minority of cases. The aim of this study was to perform a comprehensive meta-analysis of risk factors for intrahepatic (iCCA) and extrahepatic cholangiocarcinoma (eCCA), from Eastern and Western world studies. A literature search of case-control studies was performed to identify potential risk factors for iCCA and eCCA. Pooled odds ratios (ORs) with 95% CIs and heterogeneity were calculated. Funnel plots were used to assess publication bias, and meta-regression was used to select risk factors for comparison between Eastern and Western studies. A total of 13 risk factors were selected from 25 case-control studies in 7 geographically diverse countries. The strongest risk factors for both iCCA and eCCA were biliary cysts and stones, cirrhosis, hepatitis B and hepatitis C. Choledochal cysts conferred the greatest risk of both iCCA and eCCA with pooled ORs of 26.71 (95% CI 15.80-45.16) and 34.94 (24.36-50.12), respectively. No significant associations were found between hypertension and obesity for either iCCA or eCCA. Comparing Eastern and Western populations, there was a difference for the association of hepatitis B with iCCA (coefficient = -0.15195; 95% CI -0.278 to -0.025; p = 0.022). This is the most comprehensive meta-analysis of CCA risk factors to date. Some risk factors, such as diabetes, although less strong, are increasing globally and may be contributing to rising rates of this cancer. Cholangiocarcinoma (CCA) is a cancer arising in the bile ducts inside (intrahepatic CCA) and connected to the liver (extrahepatic CCA). It is a very aggressive cancer: 95% of patients die within 5 years. CCA rates are increasing globally, but the causes of CCA are poorly understood. The few risk factors that are known account for only a minority of cases. In this study, we found that the strongest risk factors for both intrahepatic and extrahepatic CCA are cysts and stones in the bile ducts, cirrhosis, and hepatitis B and C viruses. Some risk factors for CCA, such as diabetes, although less strong, are increasing globally and may be contributing to rising rates of CCA.


On the validity of memristor modeling in the neural network literature.
An analysis of the literature shows that there are two types of non-memristive models that have been widely used in the modeling of so-called "memristive" neural networks. Here, we demonstrate that such models have nothing in common with the concept of memristive elements: they describe either non-linear resistors or certain bi-state systems, which all are devices without memory. Therefore, the results presented in a significant number of publications are at least questionable, if not completely irrelevant to the actual field of memristive neural networks.


Preparation of allosamidin and demethylallosamidin photoaffinity probes and analysis of allosamidin-binding proteins in asthmatic mice.
Allosamidins, metabolites of Streptomyces with strong inhibitory activities toward family 18 chitinases, show a variety of biological activities in various organisms. We prepared photoaffinity and biotinylated probes of allosamidin and demethylallosamidin, the N-demethyl derivative that shows much stronger anti-asthmatic activity than allosamidin. Mild acid hydrolysis of allosamidins afforded mono-amine derivatives, which were amidated to prepare probes with a photoactivatable aryl azide and/or biotin moieties. The derivatives with an N-acyl group at C-2 of the D-allosamine residue at the non-reducing end of allosamidins inhibited Trichoderma chitinase as strongly as the original compounds. Since the target of allosamidins in asthma is unclear, photoaffinity probes were used to analyze allosamidin-binding proteins in bronchoalveolar lavage (BAL) fluid in IL-13-induced asthmatic mice. Ym1, a chitinase-like protein, was identified as the main allosamidin-binding protein among proteins whose expression was upregulated by IL-13 in BAL fluid. Binding of allosamidins with Ym1 was confirmed by the experiments with photoaffinity probes and recombinant Ym1.


Laparoscopic Vertical Sleeve Gastrectomy.
Bariatric surgery has emerged as the most effective means of achieving weight loss. Obesity surgery is a quickly expanding field. Laparoscopic vertical sleeve gastrectomy is a great option for patients because it is simple, exceedingly safe, has a fairly defined postoperative complication profile, and is as effective as more complex bariatric surgery options. Specific consideration of patients' comorbidities, assessment of surgeon's skill, and knowledge of preoperative, perioperative, and postoperative course is a must for all surgeons who wish to perform this procedure. If properly used, vertical sleeve gastrectomy is a powerful tool in combating obesity and its deleterious effects.


Methotrexate treatment in hand osteoarthritis refractory to usual treatments: A randomised, double-blind, placebo-controlled trial.
To examine the effect of methotrexate (MTX) on pain and structural progression in symptomatic erosive hand osteoarthritis (HOA). This 1-year prospective, single-centre, randomised, double-blind, placebo-controlled study (www.ClinicalTrial.gov, NCT01068405) followed up patients with symptomatic erosive HOA. Patients were randomised into two groups based on the drug that was administered: 10 mg methotrexate (MTX) per week or a placebo. The primary endpoint was the change in pain (determined using a visual analogue scale [VAS]) from baseline to 3 months. The secondary endpoints were pain VAS score at 12 months, clinical features (pain VAS score and function), radiographic features (the anatomical radiographic Verbruggen-Veys [VV] score and Gent University Score System), and magnetic resonance imaging (MRI) at 12 months. Sixty-four patients with HOA were randomised into either the placebo or MTX group. At 3 months, there was no significant difference in the mean decrease in the pain VAS score (mm) (MTX: 21.1 [standard deviation, 27.4], placebo: 11.7 [24.3]; p = 0.2). At 12 months, according to the VV score, erosive joints progressed significantly more to a remodelling phase in the MTX group than in the placebo group (27% vs 15%; p = 0.03). Joints with space loss appeared to be eroding less in the MTX group compared to the placebo group (8% vs 29%; p = 0.2). Synovitis on MRI at baseline could be associated with the erosive structural evolution of non-erosive joints (p = 0.02). Weekly doses of 10-mg MTX showed no superiority over the placebo in terms of pain relief at 3 or 12 months. This study was registered at www.ClinicalTrial.gov (NCT01068405).


Real-time in vivo kinetics of protoporphyrin IX after administration of 5-aminolevulinic acid in meningiomas and comparative analyses with glioblastomas.
The usefulness of 5-aminolevulinic acid (5-ALA)-mediated fluorescence-guided surgery (FGS) in meningiomas is intensely discussed. However, data about kinetics of 5-ALA and protoporphyrin (Pp) IX in meningiomas are lacking. As the first study so far, we performed longitudinal intraoperative real-time ex situ measurements of fluorescence intensity and PpIX concentrations during FGS of ten benign and two atypical meningiomas. Kinetics were subsequently compared with data from 229 glioblastomas. Spectroscopy revealed fluorescence (median 2945.65 a.u.) and PpIX accumulation (median 18.31 μg/ml) in all 43 analyzed samples. Fluorescence intensity (2961.50 a.u. vs 118.41 a.u.; p < .001) and PpIX concentrations (18.72 μg/ml vs .98 μg/ml; p < .001) were higher in samples with (N = 30) than without (N = 2) visible intraoperative tumor fluorescence. ROC curve analyses revealed a PpIX cut-off concentration of 3.85 μg/ml (AUC = .992, p = .005) and a quantitative fluorescence cut-off intensity of 286.73 a.u. (AUC = .983, p = .006) for intraoperative visible tumor fluorescence. Neither fluorescence intensity (p = .356) nor PpIX (p = .631) differed between atypical and benign meningiomas. Fluorescence and PpIX peaked 7-8 h following administration of 5-ALA. Meningiomas displayed a higher fluorescence intensity (p = .012) and PpIX concentration (p = .005) than glioblastomas 5-6 h after administration of 5-ALA. Although fluorescence was basically maintained, PpIX appeared to be cleared faster in meningiomas than in glioblastomas. Kinetics of PpIX and fluorescence intensity differ between meningiomas and glioblastomas in the early phase after 5-ALA administration. Modification of the timing of drug administration might impact visibility of intraoperative fluorescence and helpfulness of FGS and should be investigated in future analyses.


Acyclic nucleoside phosphonates with a branched 2-(2-phosphonoethoxy)ethyl chain: efficient synthesis and antiviral activity.
Series of novel acyclic nucleoside phosphonates (ANPs) with various nucleobases and 2-(2-phosphonoethoxy)ethyl (PEE) chain bearing various substituents in β-position to the phosphonate moiety were prepared. The influence of structural alternations on antiviral activity was studied. Several derivatives exhibit antiviral activity against HIV and vaccinia virus (middle micromolar range), HSV-1 and HSV-2 (lower micromolar range) and VZV and CMV (nanomolar range), although the parent unbranched PEE-ANPs are inactive. Adenine as a nucleobase and the methyl group attached to the PEE chain proved to be a prerequisite to afford pronounced antiviral activity.


Staging of Esophageal Malignancy.
Optimal treatment of esophageal cancer is a complex process dependent on many factors, including stage at diagnosis, medical fitness, physician judgment, and expertise. Despite significant advances in understanding of this cancer, survival remains low. Identifying patients with early-stage disease can enhance their outcomes dramatically. On a broader scale, staging is critical in advancing the quality of care delivered to these patients now and in the future. This article is designed to review clinicians' expertise with staging and to elaborate on the nuances frequently encountered when doing so.


Ultrasound guided vacuum aspiration treatment of caesarean scar ectopic pregnancy
To describe the case of ectopic pregnancy in the scar after caesarean section and its successful treatment using the technique of sonographically controlled vacuum aspiration. The case of a 35-year-old patient with a history of two caesarean sections referred by a district gynecologist in the 6th week of pregnancy with suspected pathological localization of pregnancy in a scar after a previous caesarean section. The procedure for the diagnosis and treatment of ectopic pregnancy in the scar after previous caesarean sections has been successfully resolved using sonographically controlled vaginal vacuum aspiration. Caesarean scar pregnancy occurs as a complication of previous caesarean section or other uterine instrumental performances. The use of the sonographically controlled vacuum aspiration technique appears to be a successful method of treating pregnancy in a scar after a caesarean section.