[The evolutionary principle in histology].
The A.A. Zavarzin's law of parallel lines in tissue evolution showing the appearance of a common tissue organization pattern is the major regularity of the cellular development of animals in phylogenesis. It carries out the metabolism principally similar in all animals in their interaction with the environment. The A.A. Zavarzin's law of parallel lines is also the main regularity of the evolution of tissue interrelations bringing about the principal processes of metabolism. The tissue evolution as well as the evolution of the organism is under the influence of factors of heredity, variability and natural selection. These factors however have an influence mediated by the organism evolution. The precess of cephalization represents a fairly important manifestation of the evolution of animals. The theory of general histogenesis reflects the main regularity of the cellular development in ontogenesis of man and animals. Being a further development of the conception of the A.A. Zavarzin's tissue evolution this theory shows the development of the animal cellular structure as a single general process of histogenesis, beginning with a zigote and continuing till the end of the organism life. The integration, heterochronia and determination manifest themselves in this process of differentiation of cells, tissues and intertissue relations.


Dependence of radiochromic film optical density post-exposure kinetics on dose and dose fractionation.
Radiochromic film (RCF) has been shown to be a precise and accurate secondary planar dosimeter for acute exposure radiation fields. However, its application to low dose-rate brachytherapy has been questioned because of possible dose-rate effects. To address this concern, we have measured the optical density (OD) of Model 55-2 RCF as a function of time (interval between the completion of irradiation and densitometry using a 633 nm laser scanner) following exposure (from less than 1 hour to 90 days) for single and split doses from 1 Gy to 100 Gy. Our work demonstrates that film darkening as a function of post-irradiation time depends significantly on total dose, with films exposed to lower doses developing faster than films given higher doses. At 1 Gy, the OD 90 days after exposure is 200% larger than that measured 1 h after exposure compared to a 20% increase over 90 days for doses larger than 20 Gy. An empirical model with time-independent, fast and slow growth terms was used to fit single exposure data. The dependence of the resulting best-fit parameters on dose was investigated. Splitting the dose into two fractions (20 Gy followed by doses of 1-80 Gy 24 h later) results in modest post-irradiation time-dependent changes in the total optical density (at most 15% at small doses), which dissipates within 20 hours following the second exposure. This experimental finding is consistent with the predictions of a simple cumulative dose superposition model. Overall, both experimental and empirical modeling suggest that dose-rate effects may be relatively small despite the strong dependence of film darkening kinetics on total dose. However, more experimental evaluation of radiochromic film response dependence on dose rate and dose-time-fractionation patterns is needed.


Surveillance Systems to Track and Evaluate Obesity Prevention Efforts.
To address the obesity epidemic, the public health community must develop surveillance systems that capture data at levels through which obesity prevention efforts are conducted. Current systems assess body mass index (BMI), diet, and physical activity behaviors at the individual level, but environmental and policy-related data are often lacking. The goal of this review is to describe US surveillance systems that evaluate obesity prevention efforts within the context of international trends in obesity monitoring, to identify potential data gaps, and to present recommendations to improve the evaluation of population-level initiatives. Our recommendations include adding environmental and policy measures to surveillance efforts with a focus on addressing underserved populations, harmonizing existing surveillance systems, including more sensitive measures of obesity outcomes, and developing a knowledgeable workforce. In addition, the widespread use of electronic health records and new technologies that allow self-quantification of behaviors offers opportunities for innovative surveillance methods.


Inhalational anaesthetics, ozone depletion, and greenhouse warming: the basics and status of our efforts in environmental mitigation.
Following their use for medicinal purposes, volatile inhalational anaesthetic agents are expelled into the atmosphere where they contribute to anthropogenic climate change. We describe recent evidence examining the benefits and harms associated with their use. The environmental harms associated with desflurane and nitrous oxide likely outweigh any purported clinical benefits. Life cycle analyses are beginning to address the many gaps in our understanding, and informing choices made on all aspects of anaesthetic care. There is, however, an urgent need to move beyond the debate about anaesthetic technique A vs. B and focus also on areas such as sustainable procurement, waste management, pharmacological stewardship and joined-up solutions. There is now compelling evidence that anaesthetists, departments and hospitals should avoid desflurane completely, and limit nitrous oxide use to settings where there is no viable alternative, as their environmental harms outweigh any perceived clinical benefit. Life cycle analyses seem supportive of total intravenous and/or regional anaesthesia. There are many other areas where choices can be made by individual anaesthetists that contribute towards reducing the environmental burden of healthcare, such as prioritising the reduction of inappropriate resource use and over-treatment. However, this all requires joined up solutions where all parts of an organisation engage.


Tranilast inhibits vascular smooth muscle cell growth and intimal hyperplasia by induction of p21(waf1/cip1/sdi1) and p53.
Tranilast, which is an antiallergic drug, has a potent effect on preventing postangioplasty restenosis. To elucidate this mechanism, we studied the effect of tranilast on the proliferation of vascular smooth muscle cells (SMCs) in vitro and in vivo. Tranilast decreased the growth rate of SMCs stimulated by either 10% FBS or platelet-derived growth factor. The IC50 value, evaluated as cell number, was 100 micromol/L. These inhibitory effects were associated with inhibition of the retinoblastoma gene product (pRb) phosphorylation. Because pRb phosphorylation is regulated by cyclin-dependent kinases (CDK), we investigated CDK2 and CDK4 activities and the expression of CDK inhibitor p21(waf1/cip1/sdi1) (p21). When SMCs were stimulated by 10% FBS or platelet-derived growth factor, CDK2 and CDK4 activities reached a maximum near the G1/S transition. Tranilast suppressed their activities by >80% without reduction of CDK2/cyclin E and CDK4/cyclin D1 protein levels. These inhibitory effects were associated with enhanced expression of p21 and elevated complexing of p21 with CDK2/CDK4. Next, rat balloon-injured carotid artery was analyzed for intimal thickening and p21 expression. Tranilast-treated rats had a 70% (P<0.001) smaller neointima/media area ratio at 14 days after balloon injury compared with the controls. Immunohistochemical staining demonstrated that, in tranilast-treated rats, p21 was already present in the neointima at day 7 and strongly expressed throughout the neointima at day 14. In control rats, p21 was not observed in the neointima at day 7 but was sparsely expressed at day 14. These data demonstrate that inhibition of CDK2/CDK4 activities by the increased expression of p21 may be one mechanism by which tranilast inhibits SMC proliferation and prevents postangioplasty restenosis.


Mutational escape from CD8+ T cell immunity: HCV evolution, from chimpanzees to man.
The mechanisms by which the hepatitis C virus (HCV) establishes persistence are not yet fully understood. Previous chimpanzee and now human studies suggest that mutations within MHC class I-restricted HCV epitopes might contribute to viral escape from cytotoxic T lymphocyte (CTL) responses. However, there are several outstanding questions regarding the role of escape mutations in viral persistence and their fate in the absence of immune selection pressure.


Could masks curtail the post-lockdown resurgence of COVID-19 in the US?
The community lockdown measures implemented in the United States from late March to late May of 2020 resulted in a significant reduction in the community transmission of the COVID-19 pandemic throughout the country. However, a number of US states are currently experiencing an alarming post-lockdown resurgence of the pandemic, triggering fears for a devastating second pandemic wave. We designed a mathematical model for addressing the key question of whether or not the universal use of face masks can halt such resurgence (and possibly avert a second wave, without having to undergo another cycle of major community lockdown) in the states of Arizona, Florida, New York and the entire US. Model calibration, using cumulative mortality data for the four jurisdictions during their respective pre-lockdown and lockdown periods, show that pre-symptomatic and asymptomatically-infectious individuals are, by far, the main drivers of the COVID-19 pandemic in each of the jurisdictions. The implication of this result is that detecting and isolating individuals with clinical symptoms of the pandemic alone (even if all of them are found) may not be sufficient to effectively curtail the pandemic. To achieve such control, it is crucially-necessary that pre-symptomatic and asymptomatically-infectious individuals are rapidly detected and isolated (and their contacts rapidly traced and tested). Our study highlights the importance of early implementation of the community lockdown measures. In particular, a sizable reduction in the burden of the pandemic would have been recorded in each of the four jurisdictions if the community lockdown measures were implemented a week or two earlier. These reductions are significantly increased if the early implementation of the lockdown measures was complemented with a public face mask use strategy. With all related control measures maintained at their baseline levels, this study shows that the pandemic would have been almost completely suppressed from significantly taking off if the lockdown measures were implemented two weeks earlier, and if a sizable percentage of the residents of the four jurisdictions wore face masks during the respective lockdown periods. The burden of the second wave of the pandemic would have been reduced significantly if the lockdown measures were extended by two weeks. We simulated the pandemic in the four jurisdictions under three levels of lifting of community lockdown, namely mild, moderate and high. For the scenario where the control measures adopted are maintained at their baseline levels during the lockdown period, our simulations show that the states of Arizona and Florida will record devastating second waves of the pandemic by the end of 2020, while the state of New York and the entire US will record milder second waves. If the community lockdown measures were lifted at the mild lifting level (i.e., only limited community contacts and business activities are allowed, in comparison to the levels of these activities allowed during the corresponding lockdown period), only the state of Florida will experience a second wave. It is further shown that the severity of the projected second waves depend on the level of lifting of the community lockdown. For instance, the projected second wave for Arizona and Florida will be more severe than their respective first waves. It is further shown that, for high level of lifting of community lockdown measures, the increased use of face masks after the lockdown period greatly reduces the burden of the pandemic in each jurisdiction. In particular, for this high lockdown lifting scenario, none of the four jurisdictions will experience a second wave if half of their residents wear face masks consistently after their respective lockdown period. A diagnostic testing strategy that increases the maximum detection rate of asymptomatic infected individuals (followed by contact tracing and self-isolation of the detected cases) greatly reduces the burden of the pandemic in all four jurisdictions, particularly if also combined with a universal face mask use strategy. Finally, it is shown that the universal use of face masks in public, with at least moderate level of compliance, could halt the post-lockdown resurgence of COVID-19, in addition to averting the potential for (and severity of) a second wave of the pandemic in each of the four jurisdictions.


Better ventilator settings using a computerized clinical tool.
The Acute Respiratory Distress Syndome (ARDS) Network low tidal volume (VT) trial paved the ground for mechanically ventilating ARDS patients with a VT of 6 mL/kg ideal body weight (IBW). Although there is no consensus that a low VT is advantageous in non-ARDS patients,it is accepted that high VT should be avoided. Because compliance rates with ventilator recommendations are 30%, there is a need for process improvement. We postulated that a computerized screen prompt that recommended VT based on height would improve compliance with low VT.During ventilator order entry, the computerized decision tool prompts the clinician and encourages ventilation of patients at 8 mL/kg IBW, and 6 mL/kg IBW for patients with ARDS. A retrospective review was performed on patients who required volume controlled mechanical ventilation over a 3-y period. Subjects were chosen randomly from the respiratory records of 6 different ICUs at a single tertiary care academic center. Half of the charts selected were before intervention of on-screen prompt, and the other half were after implementation of the computerized decision tool. The initial set VT ranged from 6.26 to 13.45 mL/kg IBW, with a mean of 8.92 mL/kg. After implementation of the on-screen prompt, mean VT decreased by 0.84 mL/kg to 8.07 mL/kg (P= .001) with a lower range of 4.73-11.56 mL/kg IBW. We also noted a significant decrease in the number of subjects placed on an initial VT > 10 mL/kg IBW from 20% to 4% (P= .003). A computerized clinical decision tool with the preferred initial VT settings based on the patients' sex and height is a safe and reliable way to increase low VT strategy compliance across multiple ICUs. Its limitations are similar to those shared by other computer-generated prompts.


Post-COVID-19 pulmonary fibrosis: An ongoing concern.
Coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 rapidly spread across the globe causing over 6 million deaths and major compromization of health facilities. The vast majority of survivors post-COVID-19 are left with variable degrees of health sequelae including pulmonary, neurological, psychological, and cardiovascular complications. Post-COVID-19 pulmonary fibrosis is one of the major concerns arising after the recovery from this pandemic. Risk factors for post-COVID-19 pulmonary fibrosis include age, male sex, and the severity of COVID-19 disease. High-resolution computed tomography provides diagnostic utility to diagnose pulmonary fibrosis as it provides more details regarding the pattern and the extent of pulmonary fibrosis. Emerging data showing similarities between post-COVID-19 pulmonary fibrosis and idiopathic pulmonary fibrosis, finding that needs further exploration. The management of post-COVID-19 pulmonary fibrosis depends on many factors but largely relies on excluding other causes of pulmonary fibrosis, the extent of fibrosis, and physiological impairment. Treatment includes immunosuppressants versus antifibrotics or both.


Commentary and historical perspective of anterior cruciate ligament rehabilitation.
This article serves as the introduction and historical perspective of anterior cruciate ligament surgery and rehabilitation. Several physician-therapist teams have been invited to share their "state of the art" techniques and to contrast their programs to that espoused by Shelbourne and Nitz in 1990. Our commentary/review of "Accelerated Rehabilitation After Anterior Cruciate Ligament Reconstruction" (Shelbourne KD, Nitz P, Am J Sports Med 18:292-299, 1990) is provided to contextualize the reader to what most clinicians would recognize as an extremely aggressive rehabilitation approach that is being popularized in the 1990s. A comparison is then presented of the rehabilitation sequence used in the MacIntosh procedures, demonstrating how early motion/functional rehabilitation was the hallmark of this type of extraarticular rehabilitation sequence and how today's pattern has evolved to follow that philosophy. Each of the teamed authors has attempted to present his surgery and rehabilitation/techniques and highlight differences between his program and that of Shelbourne and Nitz. We hope that the readers find this glimpse of the past and present helpful in formulating their rehabilitation sequences and that the future will be predicated on excellent basic science and clinical judgment. J Orthop Sports Phys Ther 1992;15(6):265-269.


Scaling trends and performance evaluation of 2-dimensional polarity-controllable FETs.
Two-dimensional semiconducting materials of the transition-metal-dichalcogenide family, such as MoS2 and WSe2, have been intensively investigated in the past few years, and are considered as viable candidates for next-generation electronic devices. In this paper, for the first time, we study scaling trends and evaluate the performances of polarity-controllable devices realized with undoped mono- and bi-layer 2D materials. Using ballistic self-consistent quantum simulations, it is shown that, with the suitable channel material, such polarity-controllable technology can scale down to 5 nm gate lengths, while showing performances comparable to the ones of unipolar, physically-doped 2D electronic devices.


Hypertension and genome-wide association studies: combining high fidelity phenotyping and hypercontrols.
Among the common complex diseases, hypertension has been particularly unlucky in the recent surge of positive results from genome-wide association studies. We summarize the evidence that would support continuing the effort in the hunt for a genetic basis for hypertension. The problems facing the genetic studies for hypertension are not unique, but phenotypic characterization, heterogeneity and high prevalence make it a special case requiring a more individualized approach. We argue that, even in the presence of a strong environmental component to hypertension risk, the common disease/common variant model is relevant for hypertension and discuss the issues involved in designing a genome-wide association study for hypertension. It is likely that the individual odds ratios for disease variants will be less than 1.3 and, although individually these effect sizes are minor, the combination of even a few such common polymorphisms can have substantial population attributable risks. The identification of hypertension gene variants should provide new insight into the disease susceptibility, progression and severity. This will lead to the identification of potential targets for lifestyle and pharmacological interventions, with the ultimate goal of improving prevention, diagnosis and treatment.


Quality of Life, Mental Health and Educational Stress of High School Students in the Northeast of Thailand.
Majority of high school students in Thailand aim to study at universities. Therefore, they spend a lot of time studying in both classrooms and tutorial classes, that could cause stress, health problems and deteriorate their Quality Of Life (QOL). However, there has been no study on these issues in Thai context. To describe the status of QOL, mental health, educational stress, well-being and determine factors associated with QOL among high school students in the Northeast of Thailand. This cross-sectional study was conducted in the Northeast of Thailand among 1,112 students of grade 10th, 11th and 12th. Multistage random sampling was used to select high schools in 5 provinces. A self-administered questionnaire was used to assess QOL, educational stress, anxiety, depression and well-being. The association between the covariates was observed by using Generalized Linear Mixed (logistic regression) Model (GLMM). The prevalence of high level of QOL was 36% (95%CI: 32.30 to 41.69); whereas, 26.18% (95% CI: 16.72 to 35.63) had high level of educational stress and 16.41% (95% CI: 2.20 to 30.71) had severe anxiety. Prevalence of depression was 18.55% (95%CI: 9.86 to 27.23) and low level of well-being was 13.41% (95% CI: 0.18 to 27.14). The factors significantly associated with high QOL were; not having depression (Adj. OR= 3.07; 95%CI: (2.23 to 4.22); p<0.001), had high level of general well-being (Adj. OR=3.19; 95% CI:1.99 to 5.09; p<0.001) and had low to moderate level of anxiety (OR=1.60; 95%CI:1.01 to 2.67). Most of the high school students had low to moderate levels of QOL, educational stress and anxiety. Depression, anxiety and general well-being had influences on QOL of high school students.


Mutation in TDRD9 causes non-obstructive azoospermia in infertile men.
Azoospermia is diagnosed when sperm cells are completely absent in the ejaculate even after centrifugation. It is identified in approximately 1% of all men and in 10%-20% of infertile males. Non-obstructive azoospermia (NOA) is characterised by the absence of sperm due to either a Sertoli cell-only pattern, maturation arrest, hypospermatogenesis or mixed patterns. NOA is a severe form of male infertility, with limited treatment options and low fertility success rates. In the majority of patients, the cause for NOA is not known and mutations in only a few genes were shown to be causative. We investigated the cause of maturation arrest in five azoospermic infertile men of a large consanguineous Bedouin family. Using whole genome genotyping and exome sequencing we identified a 4 bp deletion frameshift mutation in TDRD9 as the causative mutation with a Lod Score of 3.42. We demonstrate that the mutation results in a frameshift as well as exon skipping. Immunofluorescent staining with anti-TDRD9 antibody directed towards the N terminus demonstrated the presence of the protein in testicular biopsies of patients with an intracellular distribution comparable to a control biopsy. The mutation does not cause female infertility. This is the first report of a recessive deleterious mutation in TDRD9 in humans. The clinical phenotype recapitulates that observed in the Tdrd9 knockout mice where this gene was demonstrated to participate in long interspersed element-1 retrotransposon silencing. If this function is preserved in human, our data underscore the importance of maintaining DNA stability in the human male germ line.


Neuroendocrine consequences of traumatic brain injury.
This article attempts to summarize findings of recent publications addressing the prevalence, effects, and treatment of pituitary hormone deficiency following traumatic brain injury (TBI). A number of recent studies of TBI victims offer larger samples and much longer follow-up times. However, the prevalence of pituitary hormone deficiency continues to vary widely, underscoring the influence of patient selection, differences in endocrine testing, and patient's comorbidities and age. Growth hormone deficiency (GHD) continues to be the most frequently detected type of pituitary dysfunction. Several reports show the influence of GHD on functional outcomes of TBI victims beyond what is predicted by trauma severity. Emerging data support the notion growth hormone (GH) replacement as a useful intervention to improve symptomatology and functional outcomes among adequately selected GH-deficient patients recovering from TBI. Pituitary dysfunction is prevalent following TBI. Pituitary dysfunction seems to influence functional outcomes in some patients recovering from brain injury. Adequately selected patients could benefit from hormonal replacement.


Perceived gaps in early childhood development-related health policies in Iran: Findings from a qualitative exploratory study.
Background: Early childhood development (ECD) refers to physical, cognitive, emotional, and social development between 0 and 8 years. The aim of this study was to identify the gaps in ECD-related upstream and health policies in Iran. Methods: This qualitative study consisted of 2 main methodologies. First, a document analysis of heath and upstream policies related to ECD in Iran was done. Second, following a purposive sampling approach, 24 semi-structured interviews were conducted with policymakers, managers, academics, and service providers in the field of children health from October 2017 until June 2018 in Tehran, Iran. Also, a mixed approach was used for content and thematic analysis of the data. Results: In this study, 3 categories of gaps and weaknesses were found: (1) the inferior position of ECD in macro policies and lack of health considerations in upstream social development policies; (2) the imbalance approach to policymaking in health, lack of attention to removing inequalities, and centralized policymaking; (3) lack of effective ECD interventions and programs with regards to contextual considerations to improve children's development, and long-standing ignorance of developmental policies for 3-8 year-old children. Conclusion: ECD is a fundamental policy approach for childhood development, which requires the policymakers' deep insight to shift their focus from physical development to combating the shortcomings in children's development cycle, eg, physical, mental, social, emotional, cognitive, and spiritual aspects. Unless policymakers change their approach to decrease nonintegrated and noncomprehensive policymaking for ECD, child development will be compromised, endangering the eventual sustainability of the society.


The MEMIC is an ex vivo system to model the complexity of the tumor microenvironment.
There is an urgent need for accurate, scalable and cost-efficient models of the tumor microenvironment. Here, we detail how to fabricate and use the metabolic microenvironment chamber (MEMIC) - a 3D-printed ex vivo model of intratumoral heterogeneity. A major driver of the cellular and molecular diversity in tumors is accessibility to the blood stream. Whereas perivascular tumor cells have direct access to oxygen and nutrients, cells further from the vasculature must survive under progressively more ischemic environments. The MEMIC simulates this differential access to nutrients, allow co-culturing any number of cell types, and it is optimized for live imaging and other microscopy-based analyses. Owing to a modular design and full experimental control, the MEMIC provides insights into the tumor microenvironment that would be difficult to obtain via other methods. As proof of principle, we show that cells sense gradual changes in metabolite concentration leading to predictable molecular and cellular spatial patterns. We propose the MEMIC as a complement to standard in vitro and in vivo experiments, diversifying the tools available to accurately model, perturb and monitor the tumor microenvironment.


Reforming funding for chronic illness: Medicare-CDM.
Chronic diseases are a major challenge for the Australian health care system in terms of both the provision of quality care and expenditure, and these challenges will only increase in the future. Various programs have been instituted under the Medicare system to provide increased funding for chronic care, but essentially these programs still follow the traditional fee-for-service model. This paper proposes a realignment and extension of current Medicare chronic disease management programs into a framework that provides general practitioners and other health professionals with the necessary "tools" for high quality care planning and ongoing management, and incorporating international models of outcome-linked funding. The integration of social support services with the Medicare system is also a necessary step in providing high quality care for patients with complex needs requiring additional support.


Long term effect of alpha interferon in children with chronic hepatitis B.
The purpose of this study was to better define the long term prognosis of infection and disease in children with chronic hepatitis B treated with interferon (IFN) alpha. A total of 107 children with chronic hepatitis B who received IFN alpha for three or six months in two clinical trials were followed for a mean period of 69 (17) months. Response to treatment was defined as loss of hepatitis B e antigen (HBeAg) within 12 months after stopping treatment. A control group of 59 patients was also followed for a shorter mean time (46 (19) months). Sixteen (15%) treated children responded during therapy and 18 (17%) during post-treatment follow up; 31 (29%) non-responders lost HBeAg during subsequent years. High pretreatment levels of transaminases and a greater histological activity index were predictors of response. Kaplan-Meier estimates of cumulative HBeAg clearance rates at five years were similar between treated patients (60%) and controls (65%). After HBeAg clearance, all cases lost hepatitis B virus DNA and 94% had normal transaminase levels. Loss of hepatitis B surface antigen (HBsAg) occurred in four (25%) patients who responded during treatment but in none of the other treated or untreated patients. After five years' observation, the proportion of treated children with sustained HBeAg clearance comprised an equal number of responders and non-responders and did not differ from that observed in untreated controls, suggesting that IFN simply accelerated a spontaneous event. However, IFN significantly improved the rate of HBsAg loss in cases with more prominent disease activity who were early responders, and may be particularly useful in this type of patient.


Studies in porphyria. VI. Biosynthesis of porphyrins in mammalian skin and in the skin of porphyric patients.
Porphyrin biosynthesis in mammalian skin and in skin obtained from patients with selected types of porphyria has been studied. Cutaneous porphyrinogenesis required the precursor delta-aminolevulinic acid (ALA) which, when added to murine, rat, and human skin in vitro, was rapidly converted to porphyrins. Total porphyrin content was quantitated by fluorescence assay, and spectral studies indicated that more than 80% of the porphyrin produced was protoporphyrin. The majority of skin porphyrinogenesis occurrred in epidermis or in epidermal derivatives such as hair roots. Known inducers of hepatic delta-aminolevulinic acid synthetase (ALAS), the rate-limiting enzyme for heme biosynthesis, were not inducers when added to skin in vitro. Skin from patients with acute intermittent porphyria demonstrated a 43% decrease in cutaneous porphyrin production as compared to unaffected normals. This is consistent with the known deficiency of uroporphyrinogen synthetase that has been previously demonstrated in the liver and red blood cells of these patients. Porphyrinogenesis in skin of patients with porphyria cutanea tarda was not different from controls. These studies demonstrate that skin has the enzymatic capacity to synthesize porphyrins from added ALA and that cutaneous porphyrinogenesis from ALA is deficient in patients with acute intermittent porphyria.


Molecular identification of a taste receptor gene for trehalose in Drosophila.
The molecular nature of sweet taste receptors has not been fully explored. Employing a differential screening strategy, we identified a taste receptor gene, Tre1, that controls the taste sensitivity to trehalose in Drosophila melanogaster. The Tre1 gene encodes a novel protein with similarity to G protein-coupled seven-transmembrane receptors. Disruption of the Tre1 gene lowered the taste sensitivity to trehalose, whereas sensitivities to other sugars were unaltered. Overexpression of the Tre1 gene restored the taste sensitivity to trehalose in the Tre1 deletion mutant. The Tre1 gene is expressed in taste sensory cells. These results provide direct evidence that Tre1 encodes a putative taste receptor for trehalose in Drosophila.


Metabolic pattern of the acute phase of subarachnoid hemorrhage in a novel porcine model: studies with cerebral microdialysis with high temporal resolution.
Aneurysmal subarachnoid hemorrhage (SAH) may produce cerebral ischemia and systemic responses including stress. To study immediate cerebral and systemic changes in response to aneurysm rupture, animal models are needed. To study early cerebral energy changes in an animal model. Experimental SAH was induced in 11 pigs by autologous blood injection to the anterior skull base, with simultaneous control of intracranial and cerebral perfusion pressures. Intracerebral microdialysis was used to monitor concentrations of glucose, pyruvate and lactate. In nine of the pigs, a pattern of transient ischemia was produced, with a dramatic reduction of cerebral perfusion pressure soon after blood injection, associated with a quick glucose and pyruvate decrease. This was followed by a lactate increase and a delayed pyruvate increase, producing a marked but short elevation of the lactate/pyruvate ratio. Glucose, pyruvate, lactate and lactate/pyruvate ratio thereafter returned toward baseline. The two remaining pigs had a more severe metabolic reaction with glucose and pyruvate rapidly decreasing to undetectable levels while lactate increased and remained elevated, suggesting persisting ischemia. The animal model simulates the conditions of SAH not only by deposition of blood in the basal cisterns, but also creating the transient global ischemic impact of aneurysmal SAH. The metabolic cerebral changes suggest immediate transient substrate failure followed by hypermetabolism of glucose upon reperfusion. The model has features that resemble spontaneous bleeding, and is suitable for future research of the early cerebral and systemic responses to SAH that are difficult to study in humans.


Determination of Single-Kidney Glomerular Filtration Rate in Human Subjects by Using CT.
To test the hypothesis that computed tomography (CT)-derived measurements of single-kidney glomerular filtration rate (GFR) obtained in human subjects with 64-section CT agree with those obtained with iothalamate clearance, a rigorous reference standard. The institutional review board approved this HIPAA-compliant study, and written informed consent was obtained. Ninety-six patients (age range, 51-73 years; 46 men, 50 women) with essential (n = 56) or renovascular (n = 40) hypertension were prospectively studied in controlled conditions (involving sodium intake and renin-angiotensin blockade). Single-kidney perfusion, volume, and GFR were measured by using multidetector CT time-attenuation curves and were compared with GFR measured by using iothalamate clearance, as assigned to the right and left kidney according to relative volumes. The reproducibility of CT GFR over a 3-month period (n = 21) was assessed in patients with renal artery stenosis who were undergoing stable medical treatment. Statistical analysis included the t test, Wilcoxon signed rank test, linear regression, and Bland-Altman analysis. CT GFR values were similar to those of iothalamate clearance (mean ± standard deviation, 38.2 mL/min ± 18 vs 41.6 mL/min ± 17; P = .062). Stenotic kidney CT GFR in patients with renal artery stenosis was lower than contralateral kidney GFR or essential hypertension single-kidney GFR (mean, 23.1 mL/min ± 13 vs 36.9 mL/min ± 17 [P = .0008] and 45.2 mL/min ± 16 [P = .019], respectively), as was iothalamate clearance (mean, 26.9 mL/min ± 14 vs 38.5 mL/min ± 15 [P = .0004] and 49.0 mL/min ± 14 [P = .001], respectively). CT GFR correlated well with iothalamate GFR (linear regression, CT GFR = 0.88*iothalamate GFR, r(2) = 0.89, P < .0001), and Bland-Altman analysis was used to confirm the agreement. CT GFR was also moderately reproducible in medically treated patients with renal artery stenosis (concordance coefficient correlation, 0.835) but was unaffected by revascularization (mean, 25.3 mL/min ± 15.2 vs 30.3 mL/min ± 18.5; P = .097). CT assessments of single-kidney GFR are reproducible and agree well with a reference standard. CT can be useful to obtain minimally invasive estimates of bilateral single-kidney function in human subjects.


Aceruloplasminemia: Waiting for an Efficient Therapy.
Aceruloplasminemia is an ultra-rare hereditary disorder caused by defective production of ceruloplasmin. Its phenotype is characterized by iron-restricted erythropoiesis and tissue iron overload, diabetes, and progressive retinal and neurological degeneration. Ceruloplasmin is a ferroxidase that plays a critical role in iron homeostasis through the oxidation and mobilization of iron from stores and subsequent incorporation of ferric iron into transferrin (Tf), which becomes available for cellular uptake via the Tf receptor. In addition, ceruloplasmin has antioxidant properties preventing the production of deleterious reactive oxygen species via the Fenton reaction. Some recent findings suggest that aceruloplasminemia phenotypes can be more heterogeneous than previously believed, varying within a wide range. Within this large heterogeneity, microcytosis with or without anemia, low serum iron and high serum ferritin, and diabetes are the early hallmarks of the disease, while neurological manifestations appear 10-20 years later. The usual therapeutic approach is based on iron chelators that are efficacious in reducing systemic iron overload. However, they have demonstrated poor efficacy in counteracting the progression of neurologic manifestations, and also often aggravate anemia, thereby requiring drug discontinuation. Open questions remain regarding the mechanisms leading to neurological manifestation and development of diabetes, and iron chelation therapy (ICT) efficacy. Recent studies in animal models of aceruloplasminemia support the possibility of new therapeutic approaches by parenteral ceruloplasmin administration. In this review we describe the state of the art of aceruloplasminemia with particular attention on the pathogenic mechanisms of the disease and therapeutic approaches, both current and perspective.


A home program of strength training, movement strategy training and education did not prevent falls in people with Parkinson's disease: a randomised trial.
For people with idiopathic Parkinson's disease, does a 6-week, comprehensive, home exercise program reduce falls and disability and improve health-related quality of life? Is the program cost-effective? Randomised, controlled trial with concealed allocation and assessor blinding. One hundred and thirty-three community-dwelling adults with Parkinson's disease. The experimental group completed a 6-week home program comprising progressive resistance strength training, movement strategy training and falls education. The control group completed 6 weeks of non-specific life skills training. Participants in both groups received weekly therapist-guided sessions for 6 consecutive weeks and a weekly self-directed home program. The primary outcome was the rate of falls, documented for the 12-month period immediately after therapy. Secondary outcomes were disability and health-related quality of life, assessed before and after intervention and at a 12-month follow-up. A total of 2255 falls were reported by the 12-month follow-up. The proportion of fallers in the experimental and control groups was 61 and 72%, respectively, which was not statistically significantly different (RR=0.85, 95% CI 0.66 to 1.09). There was no significant between-group difference in the rate of falls (incidence rate ratio=1.58, 95% CI 0.73 to 3.43). A survival analysis of participant time to first fall did not show a significant between-group difference (log-rank test χ2=0.79, p=0.37). No significant between-group differences occurred for mobility, disability or quality of life. The mean cost of delivering the experimental intervention was AUD1596. A home program of strength and movement strategy training and falls education does not prevent falls when applied at the dose used in this study. Arguably, the dosage of therapy was insufficient. Future trials need to explore further therapy content, repetitions and duration, in order to optimise outcomes and cost-effectiveness. [Morris ME, Taylor NF, Watts JJ, Evans A, Horne M, Kempster P, Danoudis M, McGinley J, Martin C, Menz HB (2017) A home program of strength training, movement strategy training and education did not prevent falls in people with Parkinson's disease: a randomised trial. Journal of Physiotherapy 63: 94-100].


Valsa mali Pathogenic Effector VmPxE1 Contributes to Full Virulence and Interacts With the Host Peroxidase MdAPX1 as a Potential Target.
The Valsa canker, caused by Valsa mali (V. mali), is a destructive disease of apple in Eastern Asia. Effector proteins are important for fungal pathogenicity. We studied a candidate effector VmPxE1 isolated based on the genome information of V. mali. By using the yeast invertase secretion assay system, VmPxE1 was shown to contain a signal peptide with secretory functions. VmPxE1 can suppress BCL-2-associated X protein (BAX)-induced cell death with a high efficacy of 92% in Nicotiana benthamiana. The expression of VmPxE1 was upregulated during the early infection stage and deletion of VmPxE1 led to significant reductions in virulence on both apple twigs and leaves. VmPxE1 was also shown to target an apple ascorbate peroxidase (MdAPX1) by the yeast two-hybrid screening, bimolecular fluorescence complementation and in vivo co-immunoprecipitation. Sequence phylogenetic analysis suggested that MdAPX1 was an ascorbate peroxidase belonging to a subgroup of heme-dependent peroxidases of the plant superfamily. The ectopic expression of MdAPX1 in the mutant of VmPxE1 significantly enhanced resistance to H2O2, while the presence of VmPxE1 seems to disturb MdAPX1 function. The present results provide insights into the functions of VmPxE1 as a candidate effector of V. mali in causing apple canker.


Vaccines for canine leishmaniasis.
Leishmania infantum is the obligatory intracellular parasite of mammalian macrophages and causes zoonotic visceral leishmaniasis (ZVL). The presence of infected dogs as the main reservoir host of ZVL is regarded as the most important potential risk for human infection. Thus the prevention of canine visceral leishmaniasis (CVL) is essential to stop the current increase of the Mediterranean visceral leishmaniasis. Recently considerable advances in achieving protective immunization of dogs and several important attempts for achieving an effective vaccine against CVL lead to attracting the scientists trust in its important role for eradication of ZVL. This paper highlights the recent advances in vaccination against canine visceral leishmaniasis from 2007 until now.


[The development of the omental bursa].
The development of the omental bursa (lesser sac) was studied in serial sections of hybrid mouse embryos of 9-11 1/2 days. The anlage of the bursa is represented by several indentations on the right side of the mesenterium. Further growth and topographic alterations of the adjoining organs determine the definitive shape of the lesser sac. On the left side, the recessus pneumato-entericus is a transient structure which persists only until the 11th day of gestation. In humans, the anlage of the bursa is almost identical with its origin in mice. The bursa facilitates the separation from the digestive tract of several organs developing in the mesentery.


An emerging phenotype of central nervous system involvement in Pompe disease: from bench to bedside and beyond.
Pompe disease (PD) is a lysosomal storage disorder caused by deficiency of the lysosomal enzyme acid-alpha glucosidase (GAA). Pathogenic variants in the GAA gene lead to excessive accumulation of lysosomal glycogen primarily in the cardiac, skeletal, and smooth muscles. There is growing evidence of central nervous system (CNS) involvement in PD. Current research is focused on determining the true extent of CNS involvement, its effects on behavior and cognition, and effective therapies that would correct the disease in both muscle and the CNS. This review article summarizes the CNS findings in patients, highlights the importance of research on animal models, explores the probable success of gene therapy in reversing CNS pathologies as reported by some breakthrough preclinical studies, and emphasizes the need to follow patients and monitor for CNS involvement over time. Lessons learned from animal models (bench) and from the literature available to date on patients will guide future clinical trials in patients (bedside) with PD. Our preliminary studies in infantile PD show that some patients are susceptible to early and extensive CNS pathologies, as assessed by neuroimaging and developmental assessments. This article highlights the importance of neuroimaging which could serve as useful tools to diagnose and monitor certain CNS pathologies such as white matter hyperintense foci (WMF) in the brain. Longitudinal studies with large sample sizes are warranted at this time to better understand the emergence, progression and consequences of CNS involvement in patients with PD.


Metabolic Profiling of Green Tea Treatments in Zucker Diabetic Rats Using 1H NMR.
This study has investigated the metabolic effects of catechin-rich green tea (GT) and its formulation with ascorbic acid (AA) on the Zucker rat model of type 2 diabetes. AA is used to protect the GT catechins during digestion and increase bioavailability. Thirty two Zucker diabetic fatty (ZDF) rats were randomly divided into four groups (n=8 in each group) and treated with water, GT, AA and GT+AA respectively for five weeks. Urinary metabolic profiles were determined using 1H NMR spectroscopy. Fourteen metabolites were identified and their 24-hr excretions were quantified. Changes in the 14 metabolites demonstrated differential treatment effects on the metabolism of ZDF rats. GT and AA were found to be able to independently reduce urinary excretions of most metabolites that were over-excreted in the control diabetic rats, such as oxidative stress marker metabolites and TCA cycle metabolites. GT showed a great potential in controlling metabolic acidosis by suppressing the excretion of lactic acid and acetic acid from diabetic rats and GT+AA showed a remarkably stronger suppression than GT while AA was unable to suppress these two acids. Further investigation is needed to better understand the role of GT and/or formulated GT in altering the metabolic pathways in the diabetic animal model as well as in humans.


Impact of a clinical guidelines program for breast and colon cancer in a French cancer center.
Between 1993 and 1994, the 80 physicians in the French comprehensive cancer center, Léon Bérard, developed and implemented a Clinical Practice Guidelines (CPGs) project based on an analysis of the literature and a consensus of intrainstitutional experts. The aims of this project are to assist community-based oncologists in their decision making and to minimize inappropriate variation in practices. A study was designed to assess the impact of CPGs on management of breast and colon cancer. A "before-after" study, using institutional computerized records of patients with breast or colon cancer. Records for 100 women with localized breast cancer were randomly selected from those available in 1993 and 1995, and those for all patients newly referred with colon cancer in 1993 and 1995 (77 and 81 patients, respectively). Medical decisions on these records were analyzed to assess their compliance with the CPGs. (A systematic search of the literature was performed to determine the scientific evidence for noncompliant decisions.) Of 375 available medical decisions, 350 were assessable. The compliance rate with CPGs for breast cancer was significantly higher in 1995 compared with 1993, 54% (54/99; 95% confidence interval [CI], 44%-64%) vs 19% (18/95; 95% CI, 11%-27%) (P<.001). The compliance rate for colon cancer was also significantly higher in 1995 than in 1993, 70% (62/88; 95% CI, 60%-80%) vs 50% (34/ 68; 95% CI, 38%-62%) (P=.009). In 1993, 42% (40/95; 95% CI, 32%-52%) of medical decisions for breast cancer and in 1995, 68% (67/99; 95% CI, 59%-77%) conformed with the CPGs or were judged to be based on "scientific evidence." In 1993, 71% (48/68; 95% CI, 60%-81%) of medical decisions for colon cancer, and in 1995 81% (71/88; 95% CI, 73%-89%) conformed with the CPGs or were judged to be based on scientific evidence. Compliance rates were significantly higher in 1995 for both cancers. The development and implementation of CPGs for cancer management seem to result in significant changes in medical practice, although a causal relationship between changes and CPGs is not demonstrated in this study.


Temporal pattern of africanization in a feral honeybee population from Texas inferred from mitochondrial DNA.
The invasion of Africanized honeybees (Apis mellifera L.) in the Americas provides a window of opportunity to study the dynamics of secondary contact of subspecies of bees that evolved in allopatry in ecologically distinctive habitats of the Old World. We report here the results of an 11-year mitochondrial DNA survey of a feral honeybee population from southern United States (Texas). The mitochondrial haplotype (mitotype) frequencies changed radically during the 11-year study period. Prior to immigration of Africanized honeybees, the resident population was essentially of eastern and western European maternal ancestry. Three years after detection of the first Africanized swarm there was a mitotype turnover in the population from predominantly eastern European to predominantly A. m. scutellata (ancestor of Africanized honeybees). This remarkable change in the mitotype composition coincided with arrival of the parasitic mite Varroa destructor, which was likely responsible for severe losses experienced by colonies of European ancestry. From 1997 onward the population stabilized with most colonies of A. m. scutellata maternal origin.


[Effects of salvianolate on cardiomyocytes apoptosis and heart function in a swine model of acute myocardial infarction].
To observe the effects of salvianolate on cardiomyocytes apoptosis and heart function in a swine model of acute myocardial infarction (AMI). A total of 21 young swine were randomly divided into untreated group, low-dose salvianolate (LS) group and high-dose salvianolate (HS) group (7 in each group). AMI was induced by ligaturing left anterior descending coronary artery. After the operation, 400 or 200 mg salvianolate dissolved in 250 mL 5% glucose saline was administered by intravenous drip to swine in the HS group and the LS group respectively for 7 days. The swine in the untreated group were administered with 250 mL 5% glucose saline. The left ventricular ejection fraction (LVEF) and myocardial perfusion were measured by gated myocardial perfusion imaging at the end of the 4th week after operation. And myocardial apoptosis was detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. Apoptosis index was calculated under an optical microscope. Myocardial apoptotic index in the edge of myocardial infarction was decreased in the HS group and the LS group, which in the HS group was lower than that in the untreated group (P<0.05). Radioactive defect regions were found by gated myocardial perfusion imaging in all the three groups, but those in the HS group and the LS group were less than those in the untreated group (P<0.05). And the levels of myocardial perfusion and LVEF in the HS group were significantly higher than those in the untreated group (P<0.05). Salvianolate administered by intravenous drip can inhibit the cardiomyocytes apoptosis and improve the function of heart after AMI in swine.


[Chorea and high altitude erythrocytosis: a case report].
Chorea is a movement disorder which is rarely caused by erythrocytosis. Erithrocytosis or polycythemia is the augmentation of the absolute erythrocytic mass and its most common primary cause is polycythemia vera. Some of the secondary causes are erythrocytosis by central hipoxia occurring in people who live in the highlands. Chore is a rare neurological manifestation of erythrocytosis occurring in 0.5 to 2% of these patients. There was a report of a 71 year-old male patient with generalized chorea of sudden onset predominant in mouth, tongue and face and lower limbs, secondary to acquired erythrocytosis, which improved after the number of erythrocytes decreased through successive sessions of phlebotomy.


Development of cancer genetic services in the UK: A national consultation.
Technological advances in DNA sequencing have made gene testing fast and affordable, but there are challenges to the translation of these improvements for patient benefit. The Mainstreaming Cancer Genetics (MCG) programme is exploiting advances in DNA sequencing to develop the infrastructure, processes and capabilities required for cancer gene testing to become routinely available to all those that can benefit. The MCG programme held a consultation day to discuss the development of cancer genetics with senior representation from all 24 UK cancer genetic centres. The current service landscape and capacity for expansion was assessed through structured questionnaires. Workshop discussion addressed the opportunities and challenges to increasing cancer gene testing in the National Health Service (NHS). Services vary with respect to population served and models of service delivery, and with respect to methods and thresholds for determining risk and testing eligibility. Almost all centres want to offer more cancer gene testing (82%) and reported increasing demand for testing from non-genetic clinical colleagues (92%). Reported challenges to increasing testing include the complexity of interpreting the resulting genetic data (79%), the level of funding and complexity of commissioning (67%), the limited capacity of current processes and cross-disciplinary relationships (38%), and workforce education (29%). Priorities to address include the development and evaluation of models of increasing access to gene testing, the optimal process for interpretation of large-scale genetic data, implementation of appropriate commissioning and funding processes, and achieving national consistency. The UK cancer genetics community have high expertise and strong commitment to maximising scientific advances for improved patient benefit and should be pivotally involved in the implementation of increased cancer gene testing.


Heparin fragments induce cervical inflammation by recruiting immune cells through Toll-like receptor 4 in nonpregnant mice.
Inflammation is a hallmark in the human cervix remodelling. A possible candidate inducing the inflammatory driven ripening of the cervix is the matrix component heparan sulphate, which has been shown to be elevated in late pregnancy in the cervix and uterus. Heparin and a glycol-split low molecular weight heparin (gsHep) with low anticoagulant potency has been shown to enhance myometrial contraction and interleukin (IL)-8 production by cervical fibroblasts. The aim of this study was to investigate the mechanism by which heparin promotes cervical inflammation. Wild-type, Toll-like receptor 4 (TLR4), Myeloid differentiation primary response gene 88 (MyD88) and Interferon regulatory factor 3 (IRF3)-deficient mice were treated by deposition of gsHep into the vaginas of nonpregnant mice. To identify which cells that responded to the heparin fragments, a rhodamine fluorescent construct of gsHep was used, which initially did bind to the epithelial cells and were at later time points located in the sub-mucosa. The heparin fragments induced a strong local inflammatory response in wild-type mice shown by a rapid infiltration of neutrophils and to a lesser extent macrophages into the epithelium and the underlying extracellular matrix of the cervix. Further, a marked migration into the cervical and vaginal lumen was seen by both neutrophils and macrophages. The induced mucosal inflammation was strongly reduced in TLR4- and IRF3-deficient mice. In conclusion, our findings suggest that a TLR4/IRF3-mediated innate immune response in the cervical mucosa is induced by gsHep. This low anticoagulant heparin version, a novel TLR4 agonist, could contribute to human cervical ripening during the initiation of labour.


Sulfur and hydrogen isotope anomalies in meteorite sulfonic acids.
Intramolecular carbon, hydrogen, and sulfur isotope ratios were measured on a homologous series of organic sulfonic acids discovered in the Murchison meteorite. Mass-independent sulfur isotope fractionations were observed along with high deuterium/hydrogen ratios. The deuterium enrichments indicate formation of the hydrocarbon portion of these compounds in a low-temperature environment that is consistent with that of interstellar clouds. Sulfur-33 enrichments observed in methanesulfonic acid could have resulted from gas-phase ultraviolet irradiation of a precursor, carbon disulfide. The source of the sulfonic acid precursors may have been the reactive interstellar molecule carbon monosulfide.


Incarceration of a colonoscope in an inguinal hernia: Case report and literature review.
Incarceration of an endoscope in an inguinal hernia may occur during the course of routine colonoscopy. The incarceration may occur on insertion or withdrawal and frequently the hernia is not suspected prior to the colonoscopy. Most commonly, a left sided inguinal hernia is involved, however right inguinal hernias may be implicated in subjects with altered anatomy post abdominal surgery. Incarceration of an endoscope in an inguinal hernia has been seldom reported in the literature which is likely to be related to under reporting. A range of techniques have been suggested by various authors over the last four decades to manage this unusual complication of colonoscopy. These techniques include utilizing fluoroscopy, manual external pressure and/or the fitting of a cap onto the tip of the colonoscope to facilitate colonoscopic navigation. The authors present a case report of incarceration of the colonoscope on withdrawal in an unsuspected left inguinal hernia with a review of the literature on the management of this colonoscopic complication. A management strategy is suggested.


Evolution of a community-centred experiential learning module: A mixed-methods approach to promote social accountability and community partnership in undergraduate medical education.
This article was migrated. The article was marked as recommended. Background: Education in social determinants of health (SDH) has become an important part of medical curricula, facilitated increasingly through students' experiential learning with communities. The Community and Workplace Centred Learning Experience (CWCLE) module of the University of Saskatchewan, Canada, intends to integrate and extend second-year medical students' attitudes, skills, and knowledge about SDH and community resources. We aimed to: 1) Solicit students' self-evaluation of their ability to perform module learning objectives, 2) Assess module impact on student attitudes toward SDH, 3) Obtain feedback from community partners and students about their community experiences, and 4) Use feedback to collaboratively develop recommendations to enhance the CWCLE module. Methods: We used a mixed-method approach to combine quantitative data with stories and personal experiences. We developed an online survey for two cohorts of students who had completed the module, soliciting students to self-evaluate their perceived abilities to perform the module's learning objectives and evaluating students' attitudes towards SDH. We invited representatives from community agencies involved in the CWCLE module to participate in focus groups. We also held separate focus groups with students who participated in the online survey to elaborate on their survey comments. Results: In total, 145 students participated in the online survey (response rate=72.5%). Eleven community agency representatives and seven students participated in five focus groups. Our results demonstrate that medical students benefit from community-based experiential learning of SDH and community resources. We trace evaluations and discussions in the ongoing development of this community-based experiential learning module from its initial, primarily medical-school driven designs, towards a substantial involvement of community-based organizations in its operation and continuing redevelopment. Conclusions: Our mixed method offered us a better understanding of module impact and opportunities for improvement. This module evaluation and reform generated opportunities for community partners to influence decisions in medical education and led to a collaborative evolution of a community-centred learning experience. Medical schools should actively engage community partners in teaching behavioural and social components of the curriculum and acknowledge their partners' expertise to promote community engagement and social accountability in medical education.


Takotsubo syndrome. Case report and literature review.
Takotsubo syndrome is a cardiopathy characterized by a dyskinesia or left ventricular apical ballooning and subsequent recovery of ventricular function. We undertook this study to present a case of Takotsubo syndrome. We present the case of a 60-year-old female with a 26-year history of systemic arterial hypertension treated with 40 mg of termisartan daily. She started with headache after an emotional event (witness to an aggression of her son) associated with elevation of blood pressure that was suddenly accompanied by intense oppressive chest pain and neurovegetative symptoms. Electrocardiogram showed ST-segment elevation in anteroseptal and lateral derivations. Catheterization showed a 50% obstruction in the anterior descending artery and apical dyskinesia with hypercontractile base and 35% left ventricular ejection fraction. We performed stent angioplasty in the responsible artery. Nuclear medicine study showed extensive apical infarction, and echocardiogram agreed with transient apical dyskinesia and basal hypercontractility. After a 1-month evolution, echocardiogram mobility was completely recovered and myocardial perfusion study was normal 4 months later. We show a transitory apical ballooning secondary to acute myocardial infarction associated with a significantly stressful event that resulted in complete improvement of ventricular function with percutaneous coronary intervention during the following 4 months.


Elevated fibroblast growth factor-inducible 14 expression transforms proneural-like gliomas into more aggressive and lethal brain cancer.
High-grade gliomas (HGGs) are aggressive, treatment-resistant, and often fatal human brain cancers. The TNF-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor-inducible 14 (Fn14) signaling axis is involved in tissue repair after injury and constitutive signaling has been implicated in the pathogenesis of numerous solid cancers. The Fn14 gene is expressed at low levels in the normal, uninjured brain but is highly expressed in primary isocitrate dehydrogenase wild-type and recurrent HGGs. Fn14 signaling is implicated in numerous aspects of glioma biology including brain invasion and chemotherapy resistance, but whether Fn14 overexpression can directly promote tumor malignancy has not been reported. Here, we used the replication-competent avian sarcoma-leukosis virus/tumor virus A system to examine the impact of Fn14 expression on glioma development and pathobiology. We found that the sole addition of Fn14 to an established oncogenic cocktail previously shown to generate proneural-like gliomas led to the development of highly invasive and lethal brain cancer with striking biological features including extensive pseudopalisading necrosis, constitutive canonical and noncanonical NF-κB pathway signaling, and high plasminogen activator inhibitor-1 (PAI-1) expression. Analyses of HGG patient datasets revealed that high human PAI-1 gene (SERPINE1) expression correlates with shorter patient survival, and that the SERPINE1 and Fn14 (TNFRSF12A) genes are frequently co-expressed in bulk tumor tissues, in tumor subregions, and in malignant cells residing in the tumor microenvironment. These findings provide new insights into the potential importance of Fn14 in human HGG pathobiology and designate both the NF-κB signaling node and PAI-1 as potential targets for therapeutic intervention. MAIN POINTS: This work demonstrates that elevated levels of the TWEAK receptor Fn14 in tumor-initiating, neural progenitor cells leads to the transformation of proneural-like gliomas into more aggressive and lethal tumors that exhibit constitutive NF-κB pathway activation and plasminogen activator inhibitor-1 overexpression.


Pathophysiology of chronic rejection.
Acute rejection and chronic rejection have entirely different characteristics as immune responses. Episodes of acute rejection are characterized by the strong stimulation of T cells by alloantigen presentation and the subsequent activation and proliferation of immunocompetent antiallograft T cells. In comparison, chronic rejection appears to result as a response to ongoing, low-grade injuries to the allograft vascular endothelium. We have proposed that the parenchymal and endothelial cells in the allograft secrete growth factors in response to this irritation, leading to proliferation of smooth muscle cells and influx of myocytes into the intima. The end product is the clinically observed allograft inflammation and arteriosclerosis. The different biologic mechanisms of acute and chronic rejection demand correspondingly different methods of treatment. Recent research exploring the causes and mechanisms of chronic rejection thus emphasize the importance of several interrelated inflammatory cascades within the vascular walls of the allograft. The complexity of this immune response probably precludes the development of a single therapy for the prophylaxis or treatment of chronic rejection. The best way to control chronic rejection may be to regulate the allograft's local production of cytokines, growth factors, and eicosanoids. No current drug or treatment protocol has been proven to do this effectively. Second-generation immunosuppressants, such as mycophenolate mofetil, may prove helpful by reducing the frequency and intensity of acute rejection episodes, a primary risk factor for chronic rejection. It may be possible in the next few years to develop new treatments that would double the current 7-to 8-year half-life expected for renal transplants. This would be a significant advance in the treatment of chronic rejection.


Are ankylosing spondylitis, psoriatic arthritis and undifferentiated spondyloarthritis associated with an increased risk of cardiovascular events? A prospective nationwide population-based cohort study.
To investigate the risk of first-time acute coronary syndrome (ACS), stroke and venous thromboembolism (VTE) in patients with ankylosing spondylitis (AS), psoriatic arthritis (PsA) and undifferentiated spondyloarthritis (uSpA), compared to each other and to the general population (GP). This is a prospective nationwide cohort study. Cohorts with AS (n = 6448), PsA (n = 16,063) and uSpA (n = 5190) patients and a GP (n = 266,435) cohort, were identified 2001-2009 in the Swedish National Patient and Population registers. The follow-up began 1 January 2006, or 6 months after the first registered spondyloarthritis (SpA) diagnosis thereafter, and ended at ACS/stroke/VTE event, death, emigration or 31 December 2012. Crude and age- and sex-standardized incidence rates (SIRs) and hazard ratios (HRs) were calculated for incident ACS, stroke or VTE, respectively. Standardized to the GP cohort, SIRs for ACS were 4.3, 5.4 and 4.7 events per 1000 person-years at risk in the AS, PsA and uSpA cohort, respectively, compared to 3.2 in the GP cohort. SIRs for stroke were 5.4, 5.9 and 5.7 events per 1000 person-years at risk in the AS, PsA and uSpA cohort compared to 4.7 in the GP cohort. Corresponding SIRs for VTE were 3.6, 3.2 and 3.5 events per 1000 person-years at risk compared to 2.2 in the GP cohort. Age-and sex-adjusted HRs (95% CI) for ACS events were significantly increased in AS (1.54 (1.31-1.82)), PsA (1.76 (1.59-1.95)) and uSpA (1.36 (1.05-1.76)) compared to GP. Age-adjusted HRs for ACS was significantly decreased in female AS patients (0.59 (0.37-0.97)) compared to female PsA patients. Age-and sex-adjusted HRs for stroke events were significantly increased in AS (1.25 (1.06-1.48)) and PsA (1.34 (1.22-1.48)), and nonsignificantly increased in uSpA (1.16 (0.91-1.47)) compared to GP. For VTE the age-and sex-adjusted HRs for AS, PsA and uSpA were equally and significantly increased with about 50% compared to GP. Patients with AS, PsA and uSpA are at increased risk for ACS and stroke events, which emphasizes the importance of identification of and intervention against cardiovascular risk factors in SpA patients. Increased alertness for VTE is warranted in patients with SpA.


[Characteristics of foot blood supply in rigidity of the great toe].
By means of angiography, made in 20 patients with rigidity of the first toes due to osteoarthrosis, there have been revealed chronic deficiency of blood circulation in the foot tissues, especially in its antero-internal section. Blood circulation deficiency is due either to the congenital anomaly of the foot vessels or to the consequences of their traumatic injuries. Blood supply deficiency results in degenerative-dystrophic changes of the first metatarsophalangeal articulations with development of their rigidity, aggravated by post-ischemic contracture of the foot muscles.


A Statistical Assessment of Saturation and Mobile Sampling Strategies to Estimate Long-Term Average Concentrations across Urban Areast.
The objectives of this study were: (1) to quantify the errors associated with saturation air quality monitoring in estimating the long-term (i.e., annual and 5 yr) mean at a given site from four 2-week measurements, once per season; and (2) to develop a sampling strategy to guide the deployment of mobile air quality facilities for characterizing intraurban gradients of air pollutants, that is, to determine how often a given location should be visited to obtain relatively accurate estimates of the mean air pollutant concentrations. Computer simulations were conducted by randomly sampling ambient monitoring data collected in six Canadian cities at a variety of settings (e.g., population-based sites, near-roadway sites). The 5-yr (1998-2002) dataset consisted of hourly measurements of nitric oxide (NO), nitrogen dioxide (NO2), oxides of nitrogen (NOx), sulfur dioxide (SO2), coarse particulate matter (PM10), fine particulate matter (PM2.5), and CO. The strategy of randomly selecting one 2-week measurement per season to determine the annual or long-term average concentration yields estimates within 30% of the true value 95% of the time for NO2, PM10 and NOx. Larger errors, up to 50%, are expected for NO, SO2, PM2.5, and CO. Combining concentrations from 85 random 1-hr visits per season provides annual and 5-yr average estimates within 30% of the true value with good confidence. Overall, the magnitude of error in the estimates was strongly correlated with the variability of the pollutant. A better estimation can be expected for pollutants known to be less temporally variable and/or over geographic areas where concentrations are less variable. By using multiple sites located in different settings, the relationships determined for estimation error versus number of measurement periods used to determine long-term average are expected to realistically portray the true distribution. Thus, the results should be a good indication of the potential errors one could expect in a variety of different cities, particularly in more northern latitudes.


Deciphering the biochemical regulation of metabolic pathways in abating stress-induced damages during co-exposure of arsenic and fluoride in a non-aromatic and an aromatic rice variety.
The aim of the current work was to decipher the systemic damage and biochemical defense machinery due to combined arsenic (5 mg L-1 Na3AsO4) and fluoride (50 mg L-1 NaF) stress in two rice cultivars viz., IR-64 (non-aromatic) and Gobindobhog (aromatic), grown for 14 days, under 16/8 h light/dark photoperiodic cycle at 32 °C. Higher accumulation of arsenic and fluoride in Gobindobhog generated higher levels of H2O2 that caused higher electrolyte leakage, along with malondialdehyde and methylglyoxal formation. Higher oxidative damages severely compromised seed germination and led to chlorophyll loss, inhibition of root and shoot growth and fresh and dry weight of the seedlings. On the contrary, oxidative damage was less pronounced in IR-64, as compared to that of Gobindobhog, which can be attributed to higher accumulation of protective metabolites, i.e., osmolytes and antioxidants. Higher levels of osmolytes (proline, glycine betaine and amino acids) in IR-64 helped in maintaining the osmotic balance of the cells and the integrity of the cell membrane. Additionally, up regulated activity of enzymatic antioxidants (catalase, superoxide dismutase, ascorbate peroxidase, glutathione peroxidase and glutathione S-transferase) along with elevated levels of non-enzymatic antioxidants (flavonoids, phenolics, xanthophylls and carotenoids) played a pivotal role in controlling oxidative damages and strengthening the defense machinery in IR-64, as compared to that of Gobindobhog where lesser enhancement in the level of the above mentioned protective metabolites was noted. The present work illustrated differential phytotoxicity in rice seedlings and elucidated the yet uncharacterized biohazard associated with arsenic and fluoride co-contamination, with better adaptive features of IR-64, compared to Gobindobhog, which appeared as the sensitive variety.


[Azathioprine and 5-ASA in the prevention of postoperative recurrence of Crohn's disease].
Recurrence of Crohn's disease (CD) lesions in the neo-ileum after apparently curative resection frequently occurs after surgery. The most appropriate prophylactic treatment has not been clearly defined. This study evaluated the efficacy of 5-ASA and azathioprine in decreasing postoperative recurrence and analysed the presence of variables associated with recurrence. Thirty-nine patients (mean age 32.8 years, range 18-61) with a history of ileal or ileocecal surgical resection were studied. They received 5-ASA (3 mg/day) or azathioprine (50 mg/day) immediately after the operation and for 2 years thereafter. Patients were followed clinically (Crohn's disease activity index) and serologically every 3 months and by imaging methods every 6 months. The latter included colonoscopy with ileoscopy and if not available, small bowel barium or ultrasonographic study. Laboratory tests included ESR, C-reactive protein, white blood cell and platelet count, fibrinogen and albumin. The end-point evaluated included clinical recurrence (CR), serological recurrence (SR: alteration of at least three of the above-mentioned variables) and morphologic recurrence (MR: endoscopic recurrence > 1 according to Rutgeerts score or radiological or ultrasonographic recurrence). Eighteen patients received azathioprine and 21 received 5-ASA. Thirty-four patients were evaluated. The cumulative proportion of patients with recurrence was 29% (CR), 35% (SR) and 50% (MR). Statistical analysis did not show significant differences between the two groups. Twenty-seven patients completed the 2-year study (11 in the azathioprine group and 16 in the 5-ASA group). Crude relapse rates were 37% (CR), 44% (SR) and 69% (MR) in the 5-ASA group and 36% (CR), 45% (SR) and 64% (MR) in the azathioprine group. No statistically significant differences were observed between groups. No variables associated with recurrence were detected. In conclusion, treatment does not prevent a high percentage of postsurgical recurrence. 5-ASA (3 g/day) and azathioprine (50 mg/day) showed similar efficacy in the prevention of recurrence.


Prescribing safe supply: ethical considerations for clinicians.
The COVID-19 pandemic has exacerbated the drug poisoning epidemic in a number of ways: individuals use alone more often, there is decreased access to harm reduction services and there has been an increase in the toxicity of the unregulated drug supply. In response to the crisis, clinicians, policy makers and people who use drugs have been seeking ways to prevent the worst harms of unregulated opioid use. One prominent idea is safe supply. One form of safe supply enlists clinicians to prescribe opioids so that people have access to drugs of known composition and strength. In this paper, we assess the ethical case for clinicians providing this service. As we describe, there is much that is unknown about safe supply. However, given the seriousness of the overdose death epidemic and the current limited evidence for safe supply's efficacy, we argue that it is ethically permissible for clinicians to begin prescribing opioids for some select patients.


Resistance to Propofol Induction in a Patient Taking Modafinil: A Case Report.
After sedation with midazolam, induction of anesthesia with propofol was attempted in a patient taking modafinil. However, even after administration of a total of 6 mg/kg propofol IV, the patient continued to respond to tactile stimulation. Concurrently, the bispectral index was 72. Subsequent administration of low concentration sevoflurane by facemask induced an anesthetic depth that allowed unproblematic insertion of a laryngeal mask airway. Anesthesia for ophthalmologic surgery was maintained with sevoflurane. Modafinil may have caused resistance to propofol because of its effect on neural pathways that activate consciousness. The concentration of sevoflurane required to induce or maintain anesthesia remained unaltered.


Mycobacterium indicus pranii and Mycobacterium bovis BCG lead to differential macrophage activation in Toll-like receptor-dependent manner.
Mycobacterium indicus pranii (MIP) is an atypical mycobacterial species possessing strong immunomodulatory properties. It is a potent vaccine candidate against tuberculosis, promotes Th1 immune response and protects mice from tumours. In previous studies, we demonstrated higher protective efficacy of MIP against experimental tuberculosis as compared with bacillus Calmette-Guérin (BCG). Since macrophages play an important role in the pathology of mycobacterial diseases and cancer, in the present study, we evaluated the MIP in live and killed form for macrophage activation potential, compared it with BCG and investigated the underlying mechanisms. High levels of tumour necrosis factor-α, interleukin-12p40 (IL-12p40), IL-6 and nitric oxide were produced by MIP-stimulated macrophages as compared with BCG-stimulated macrophages. Prominent up-regulation of co-stimulatory molecules CD40, CD80 and CD86 was also observed in response to MIP. Loss of response in MyD88-deficient macrophages showed that both MIP and BCG activate the macrophages in a MyD88-dependent manner. MyD88 signalling pathway culminates in nuclear factor-κB/activator protein-1 (NF-κB/AP-1) activation and higher activation of NF-κB/AP-1 was observed in response to MIP. With the help of pharmacological inhibitors and Toll-like receptor (TLR) -deficient macrophages, we observed the role of TLR2, TLR4 and intracellular TLRs in MIP-mediated macrophage activation. Stimulation of HEK293 cells expressing TLR2 in homodimeric or heterodimeric form showed that MIP has a distinctly higher level of TLR2 agonist activity compared with BCG. Further experiments suggested that TLR2 ligands are well exposed in MIP whereas they are obscured in BCG. Our findings establish the higher macrophage activation potential of MIP compared with BCG and delineate the underlying mechanism.


Evidence that platelet and tumour heparanases are similar enzymes.
In order to enter tissues, blood-borne metastatic tumour cells and leucocytes need to extravasate through the vascular basal lamina (BL), a process which involves a battery of degradative enzymes. A key degradative enzyme is the endoglycosidase heparanase, which cleaves heparan sulphate (HS), an important structural component of the vascular BL. Previously, tumour-derived heparanase activity (which has been shown to be related to the metastatic potential of murine and human melanoma cell lines) was reported to cleave HS and be inhibited by heparin, as distinct from human platelet heparanase, which cleaved both substrates [Nakajima, Irimura and Nicolson (1988) J. Cell Biochem. 36, 157-167]. We recently reported the purification of human platelet heparanase and showed that the enzyme is a 50-kDa endoglucuronidase [Freeman and Parish (1998) Biochem. J. 330, 1341-1350]. We now report the purification and characterization of heparanase activity from highly metastatic rat 13762 MAT mammary adenocarcinoma and human HCT 116 colonic carcinoma cells and from rat liver using essentially the same procedure that was reported for purification of the human platelet enzyme. The rat 13762 MAT tumour enzyme, which has a native M(r) of 45 kDa when analysed by gel-filtration chromatography and by SDS/PAGE, was observed to be an endoglucuronidase that degraded heparin and HS to fragments of the same sizes as the human platelet enzyme does. N-deglycosylation of both the human platelet and rat 13762 MAT tumour enzymes gave, in each case, a 41-kDa band by SDS/PAGE analysis, demonstrating that the observed difference in M(r) between the platelet and tumour enzymes may have been due largely to differences in the relative amounts of N-glycosylation. Two peptides were isolated following Endoproteinase Lys-C digestion of both the human platelet and rat 13762 MAT tumour heparanases and were shown to be highly similar. Both the rat liver and human colonic carcinoma heparanases also degraded both heparin and HS to fragments of the same sizes as the human platelet enzyme does. Western-blot analysis of an SDS/PAGE gel using antibodies raised against human platelet heparanase demonstrated that human platelet, human tumour and rat tumour heparanases were immunochemically cross-reactive. In conclusion, because of the similarities in their sizes, substrate specificities, peptide sequences and immunoreactivities, we propose that heparanase activities present in human platelets, rat liver and in rat and human tumour cells are, in fact, mediated by a similar enzyme.


Transport and sorting of the solanum tuberosum sucrose transporter SUT1 is affected by posttranslational modification.
The plant sucrose transporter SUT1 from Solanum tuberosum revealed a dramatic redox-dependent increase in sucrose transport activity when heterologously expressed in Saccharomyces cerevisiae. Plant plasma membrane vesicles do not show any change in proton flux across the plasma membrane in the presence of redox reagents, indicating a SUT1-specific effect of redox reagents. Redox-dependent sucrose transport activity was confirmed electrophysiologically in Xenopus laevis oocytes with SUT1 from maize (Zea mays). Localization studies of green fluorescent protein fusion constructs showed that an oxidative environment increased the targeting of SUT1 to the plasma membrane where the protein concentrates in 200- to 300-nm raft-like microdomains. Using plant plasma membranes, St SUT1 can be detected in the detergent-resistant membrane fraction. Importantly, in yeast and in plants, oxidative reagents induced a shift in the monomer to dimer equilibrium of the St SUT1 protein and increased the fraction of dimer. Biochemical methods confirmed the capacity of SUT1 to form a dimer in plants and yeast cells in a redox-dependent manner. Blue native PAGE, chemical cross-linking, and immunoprecipitation, as well as the analysis of transgenic plants with reduced expression of St SUT1, confirmed the dimerization of St SUT1 and Sl SUT1 (from Solanum lycopersicum) in planta. The ability to form homodimers in plant cells was analyzed by the split yellow fluorescent protein technique in transiently transformed tobacco (Nicotiana tabacum) leaves and protoplasts. Oligomerization seems to be cell type specific since under native-like conditions, a phloem-specific reduction of the dimeric form of the St SUT1 protein was detectable in SUT1 antisense plants, whereas constitutively inhibited antisense plants showed reduction only of the monomeric form. The role of redox control of sucrose transport in plants is discussed.


[Circadian rhythm of biosynthetic activity of the epiphysis in relatively wild and domesticated silver foxes].
Circadian dynamics of biosynthetic activity in pineal glands of adult relatively wild and domesticated silver fox females was studied beyond the reproductive season using radioimmune and fluorometric methods. The level of melatonin, the principal pineal hormone; activities of enzymes controlling its biosynthesis; the level of its precursor and one of its metabolites, as well as those of neurotransmitters involved in the regulation of biosynthesis exhibited more or less pronounced circadian changes. The concentration of melatonin in the pineal gland at night was considerably higher in domesticated foxes than in relatively wild ones. The most likely reason for the elevated concentration of melatonin at night is its slow secretion from the pineal glands of domesticated foxes, because its concentration in plasma does not differ from that in relatively wild animals. The results obtained suggest that selection for domestic behavior has affected the adrenergic mechanism of regulation of pineal gland rhythms, as well as other functions of sympathetic divisions of the central nervous system.


Near-UV mutagenesis: photoreactivation of 365-nm-induced mutational lesions in Escherichia coli WP2s.
Reversion to tryptophan independence induced by 365-nm and 254-nm radiation was studied in Escherichia coli WP2s (B/r trp uvrA). Under aerobic conditions, the mutant frequency responses was of the fluence-square or "two-hit" type at both 365 and 254 nm when revertants were assayed on minimal agar supplemented with 2% nutrient broth (SEM plates). In contrast, when mutants were assayed on minimal agar supplemented with tryptophan only, the revertant yield was reduced to very low values at 365 nm, whereas values substantially greater than with SEM plates were obtained at 254 nm. Premutational lesions induced by both 365-nm and 254-nm radiation were photoreactivated more than 10-fold when assayed on SEM plates, implicating pyrimidine dimers as premutational lesions at both wavelengths. The strong photoreactivation of 365-nm-induced mutagenesis contrasted strikingly with the complete absence of photoreactivation of 365-nm-induced lethality in this strain.


Actomyosin stress fiber mechanosensing in 2D and 3D.
Mechanotransduction is the process through which cells survey the mechanical properties of their environment, convert these mechanical inputs into biochemical signals, and modulate their phenotype in response. These mechanical inputs, which may be encoded in the form of extracellular matrix stiffness, dimensionality, and adhesion, all strongly influence cell morphology, migration, and fate decisions. One mechanism through which cells on planar or pseudo-planar matrices exert tensile forces and interrogate microenvironmental mechanics is through stress fibers, which are bundles composed of actin filaments and, in most cases, non-muscle myosin II filaments. Stress fibers form a continuous structural network that is mechanically coupled to the extracellular matrix through focal adhesions. Furthermore, myosin-driven contractility plays a central role in the ability of stress fibers to sense matrix mechanics and generate tension. Here, we review the distinct roles that non-muscle myosin II plays in driving mechanosensing and focus specifically on motility. In a closely related discussion, we also describe stress fiber classification schemes and the differing roles of various myosin isoforms in each category. Finally, we briefly highlight recent studies exploring mechanosensing in three-dimensional environments, in which matrix content, structure, and mechanics are often tightly interrelated. Stress fibers and the myosin motors therein represent an intriguing and functionally important biological system in which mechanics, biochemistry, and architecture all converge.


A rapid and sensitive fluorometric screening assay using YO-PRO-1 to quantify tumour cell invasion through Matrigel.
A new quantitative assay for the study of tumour cell invasion in vitro is described. Employing the novel fluorescent dye YO-PRO-1, cells that penetrate Matrigel-coated transwells are counted on the basis of dye-bound cellular nucleic acid content. Following transmigration, the cells in the lower compartments are lysed by freezing in water. After a brief incubation with YO-PRO-1, nucleic acid or DNA content is measured as fluorescence intensity in 96-well microplates and quantitated by a cell- or DNA-calibration curve. Using standard curves, a linear relationship between fluorescence intensity and cell number was found in the range tested (from 100 to 80 000 cells). The mean relative intra- and inter-assay variability of the cell quantitation in this range was 3.5 and 4.2%, respectively. When applied to Matrigel invasion studies, as few as 400 cells could be counted. The quantitation could be performed within 3 h. HCT 116, MDA MB 231 and HT 29 cells were investigated as examples of tumour cells with different invasive abilities in the 48-h Matrigel invasion assay. Using YO-PRO-1, 6.5 +/- 0.6% invasive HCT 116 cells and 52.6 +/- 4.5% MDA MB 231 cells (percentage of the inoculated cell population) were measured. HT 29 cells were practically non-invasive. These results were confirmed by visual scoring of DAPI-stained nuclei. In conclusion, the main advantages of the assay are its sensitive, reproducible and rapid quantitation of tumour cell invasion in vitro and the applicability to extended sample numbers by measuring in 96-well microplates.


The aryl hydrocarbon receptor is a regulator of cigarette smoke induction of the cyclooxygenase and prostaglandin pathways in human lung fibroblasts.
Cigarette smoking can lead to chronic lung inflammation and lung cancer. Chronic inflammation, associated with expression of cyclooxygenase-2 (COX-2) and prostaglandins, predisposes to malignancy. We recently demonstrated that human lung fibroblasts are activated by cigarette smoke to express COX-2 and prostaglandin E(2) (PGE(2)). Little is known about the mechanism whereby smoke activates human lung fibroblasts to produce proinflammatory mediators. Herein, we report the central role of the aryl hydrocarbon receptor (AHR) in cigarette smoke extract (CSE)-induced COX-2, microsomal PGE(2) synthase (mPGES), and PGE(2) production in human lung fibroblasts. Western blot analysis revealed that primary strains of human lung fibroblasts express AHR and aryl hydrocarbon nuclear translocator protein, supporting the possibility that smoke activates lung fibroblasts through this pathway. Experiments were subsequently performed to determine whether the AHR was activated by CSE. Immunocytochemistry and EMSA analysis revealed that CSE induced nuclear translocation of the AHR in human lung fibroblasts. CSE decreased protein levels of the AHR, consistent with AHR ligand-induced proteosome-mediated degradation. CSE also induced mPGES-1 and COX-2 protein and increased PGE(2) production. Treatment of human fibroblasts with AHR antagonists in the presence of CSE inhibited AHR nuclear translocation as well as COX-2, mPGES-1, and PGE(2) production. These data indicate that the AHR pathway plays an important role in cigarette smoke-mediated COX-2 and PG production in human lung fibroblasts and may contribute to tobacco-associated inflammation and lung disease.


A novel point-of-care system for high-speed real-time polymerase chain reaction testing for epidermal growth factor receptor mutations in bronchial lavage fluids after transbronchial biopsy in patients with non-small cell lung cancer.
Epidermal growth factor receptor (EGFR) gene mutation testing is essential for choosing appropriate treatment options in patients with advanced non-small cell lung cancer (NSCLC). However, a time delay occurs between histological diagnosis and molecular diagnosis in clinical situations. To minimize this delay, we developed a novel point-of-care test for EGFR mutations, based on a high-speed real-time polymerase chain reaction (PCR) system designated here as ultrarapid PCR combined with highly accurate bronchoscopic sampling. We investigated whether our system for detecting EGFR mutations was valid by comparing test results with those obtained using a commercialized EGFR mutation test. We obtained small amounts of bronchial lavage fluids after transbronchial biopsies (TBBs) were performed on enrolled patients (n=168) who underwent endobronchial ultrasonography using a guide sheath (EBUS-GS). EGFR mutation analysis was performed by ultrarapid PCR immediately after EBUS-GS-TBBs were obtained (on the same day). After pathological diagnoses of NSCLC, EGFR mutation status in formalin-fixed, paraffin- embedded samples was confirmed by the PCR-invader method, and the concordance rates between the PCR methods were compared. The total diagnostic yield of EBUS-GS-TBB was 91.0%. The positive concordance rates for detecting 19del and L858R with the ultrarapid PCR and PCR-invader methods were both 100%. Negative concordance rates were 97.2 and 98.1%, respectively. We also demonstrated a dramatic effect of early erlotinib administration, based on ultrarapid PCR results, for a 52-year-old woman suffering from respiratory failure due to severe intrapulmonary metastases with poor performance status. In conclusion, ultrarapid PCR combined with EBUS-GS-TBB enabled rapid and reliable point-of-care testing for EGFR mutations.


Variable Persister Gene Interactions with (p)ppGpp for Persister Formation in Escherichia coli.
Persisters comprise a group of phenotypically heterogeneous metabolically quiescent bacteria with multidrug tolerance and contribute to the recalcitrance of chronic infections. Although recent work has shown that toxin-antitoxin (TA) system HipAB depends on stringent response effector (p)ppGppin persister formation, whether other persister pathways are also dependent on stringent response has not been explored. Here we examined the relationship of (p)ppGpp with 15 common persister genes (dnaK, clpB, rpoS, pspF, tnaA, sucB, ssrA, smpB, recA, umuD, uvrA, hipA, mqsR, relE, dinJ) using Escherichia coli as a model. By comparing the persister levels of wild type with their single gene knockout and double knockout mutants with relA, we divided their interactions into five types, namely A "dependent" (dnaK, recA), B "positive reinforcement" (rpoS, pspF, ssrA, recA), C "antagonistic" (clpB, sucB, umuD, uvrA, hipA, mqsR, relE, dinJ), D "epistasis" (clpB, rpoS, tnaA, ssrA, smpB, hipA), and E "irrelevant" (dnaK, clpB, rpoS, tnaA, sucB, smpB, umuD, uvrA, hipA, mqsR, relE, dinJ). We found that the persister gene interactions are intimately dependent on bacterial culture age, cell concentrations (diluted versus undiluted culture), and drug classifications, where the same gene may belong to different groups with varying antibiotics, culture age or cell concentrations. Together, this study represents the first attempt to systematically characterize the intricate relationships among the different mechanisms of persistence and as such provide new insights into the complexity of the persistence phenomenon at the level of persister gene network interactions.


Improved Noninterferometric Test of Collapse Models Using Ultracold Cantilevers.
Spontaneous collapse models predict that a weak force noise acts on any mechanical system, as a consequence of the collapse of the wave function. Significant upper limits on the collapse rate have been recently inferred from precision mechanical experiments, such as ultracold cantilevers and the space mission LISA Pathfinder. Here, we report new results from an experiment based on a high-Q cantilever cooled to millikelvin temperatures, which is potentially able to improve the current bounds on the continuous spontaneous localization (CSL) model by 1 order of magnitude. High accuracy measurements of the cantilever thermal fluctuations reveal a nonthermal force noise of unknown origin. This excess noise is compatible with the CSL heating predicted by Adler. Several physical mechanisms able to explain the observed noise have been ruled out.