Efficacy of pasireotide in controlling severe hypercortisolism until cardiac transplantation.
Cushing's syndrome is associated with increased morbidity and mortality. Although surgery is the first-line treatment, drugs can still play a role as an ancillary treatment to be employed while waiting for surgery, after unsuccessful operation or in patients unsuitable for surgery. We were asked to evaluate a 32-year-old male waiting for cardiac transplantation. Idiopathic hypokinetic cardiomyopathy had been diagnosed since 6 years. He was on treatment with multiple drugs, had a pacemaker, an implantable cardioverter and an external device for the support of systolic function. Physical examination showed severely impaired general status, signs of hypercortisolism and multiple vertebral compression fractures. We administered teriparatide, and the few evaluable parameters supported the diagnosis of ACTH-dependent hypercortisolism: serum cortisol was 24.2 µg/dL in the morning and 20.3 µg/dL after overnight 1 mg dexamethasone, urinary free cortisol (UFC) was 258 µg/24 h and ACTH 125 pg/mL. Pituitary CT was negative. Pasireotide 300 µg bid was administered and uptitrated to 600 µg bid. Treatment was well tolerated, achieving dramatic improvement of clinical picture with progressive normalization of serum cortisol and ACTH levels as well as UFC. After 4 months, the patient underwent successful heart transplantation. Many complications ensued and were overcome. Pituitary MRI was negative. On pasireotide 300 µg bid and prednisone 2.5 mg/day (as part of immunosuppressive therapy), morning serum cortisol and ACTH were 15.6 µg/dL and 54 pg/mL respectively, UFC was 37 µg/24 h, fasting glucose: 107 mg/dL and HbA1c: 6.5%. In conclusion, primary treatment with pasireotide achieved remission of hypercortisolism, thus allowing the patient to undergo heart transplantation. Untreated Cushing's syndrome is associated with ominous prognosis.First-line treatment is surgery (at pituitary or adrenal, according to disease localization).A few drugs are available to treat hypercortisolism.Pasireotide is a multi-ligand somatostatin analog approved for treatment of hypercortisolism.Primary treatment with pasireotide was effective in a patient with severe Cushing's syndrome, allowing him to undergo heart transplantation.


Maternal behaviour in domestic dogs.
Mammalian parental care, in most of the cases, is given by the female, who provides food, warmth, and protection. In domestic dogs, maternal behaviour shown by the dam mainly consists of contact, nursing, grooming/licking, play, punishment, thermoregulation, and motion. Peer-reviewed literature published between 1952 and 2018 was retrieved from CAB Abstracts, PubMed, ISI Web of Knowledge, Scopus and book chapters. Keywords for this search included the following terms: behaviour, bonding, altricial, precocial, offspring, maternal, whelping, nursing, domestic dogs, female dog, aggression, puppies, anogenital licking. In this review, we reported and discussed scientific information about maternal behaviour in domestic bitches, comparing altricial vs precocial species; the importance of the bonding, grooming/licking and nursing, and their impacts on puppies' behaviour; altered maternal behaviours such as aggression, cannibalism, rejection, and also the relation between hormones and maternal care behaviours. We concluded that the level of interactions between the dam and the puppies influences the physiological, cognitive and behavioural development of the litter, and the main hormones in the bitch for inducing maternal care behaviours are estradiol, oxytocin, prolactin and progesterone.


Clinical outcome of severe head injury in different protocol-driven therapies.
In the past 5 years cerebral perfusion pressure (CPP) management has become mainstream in the treatment of severe head injuries. The American Association of Neurological Surgeons guidelines (2000) suggest that CPP should be maintained at least 70 mmHg; however, there is still debate about optimal CPP level. The purpose of this study is to evaluate the effectiveness of three widely used therapies: intracranial pressure (ICP)-targeted therapy, CPP-targeted therapy with CPP > 70 mmHg, and modified CPP-targeted therapy with CPP > 60 mmHg. The clinical procedures, complications, and patient outcomes are compared. Data including patient age, sex, initial Glasgow Coma Score (GCS), ICP, CPP, fluid status, amount of mannitol and vasopressor used, daily intake and output, complications, and clinical results were collected from 213 patients with severe head injuries over a 12-year period. Patients were categorized into three groups (ICP, CPP, modified CPP [mCPP]) according to treatment protocol used. Retrospective data collection was by chart review. The mortality rate was 28.6%, 14.3%, and 13.5% in groups ICP, CPP, and mCPP, respectively. Highest intake/output ratio, amount of vasopressor used, and pulmonary complication rates were seen in group CPP patients. Group mCPP patients showed the best clinical outcome and lowest complication rate. Though CPP-targeted therapy is the most recommended therapeutic protocol, our data showed that the outcome is as good in the mCPP-targeted group with CPP > 60 mmHg as in the CPP-targeted group, but complications are fewer in the mCPP group.


Macroautophagy Proteins Control MHC Class I Levels on Dendritic Cells and Shape Anti-viral CD8(+) T Cell Responses.
The macroautophagy machinery has been implicated in MHC class II restricted antigen presentation. Here, we report that this machinery assists in the internalization of MHC class I molecules. In the absence of the autophagy factors Atg5 and Atg7, MHC class I surface levels are elevated due to decreased endocytosis and degradation. Internalization of MHC class I molecules occurs less efficiently if AAK1 cannot be recruited via Atg8/LC3B. In the absence of Atg-dependent MHC class I internalization, dendritic cells stimulate CD8(+) T cell responses more efficiently in vitro and in vivo. During viral infections, lack of Atg5 results in enhanced influenza- and LCMV-specific CD8(+) T cell responses in vivo. Elevated influenza-specific CD8(+) T cell responses are associated with better immune control of this infection. Thus, the macroautophagy machinery orchestrates T cell immunity by supporting MHC class II but compromises MHC class I restricted antigen presentation.


Gender differences in associations between cancer-related problems and relationship dissolution among cancer survivors.
Research suggests that a cancer diagnosis predicts marital dissolution more strongly for women survivors than men, but there is a paucity of research on potential processes underlying this vulnerability. The present cross-sectional study examined whether specific cancer-related problems were associated with the odds of relationship breakup following diagnosis and whether these relationships differed between male and female cancer survivors. A national cross-sectional quality of life study assessed self-reported cancer-related problems and relationship change among survivors who were either 2, 6, or 10 years post-diagnosis (n = 6099). Bivariate analyses indicated that cancer-related problems (e.g., emotional distress) were greater for divorced/separated survivors compared to those with intact relationships and were greater for women versus men. Logistic regressions indicated that for both male and female survivors, lower income, younger age, and longer time since diagnosis were associated with greater odds of divorce or separation after diagnosis (ORs > 2.14, p < .01). For women only, greater emotional distress (OR = 1.14, p < 0.01) and employment and financial problems (OR = 1.23, p < 0.0001) were associated with greater odds of post-diagnosis divorce or separation. For men only, fear of cancer recurrence was associated with greater odds of divorce or separation (OR = 1.32, p < 0.001). Female and male survivors differed in the extent to which emotional or financial/employment problems attributed to the cancer diagnosis were associated with the likelihood of reporting relationship dissolution. Although directions of causality could not be ascertained, results suggest the possibility that helping male and female cancer survivors cope with specific cancer-related problems may benefit the quality and stability of their relationships with significant others following diagnosis.


Fabrication and integration of a low-cost 3D printing-based glucose biosensor for bioprinted liver-on-a-chip.
In the last two decades, significant progress has been made in the development of more physiologically relevant organ-on-a-chip (OOC) systems that can mimic tissue microenvironments. Despite the advantages of these microphysiological systems, such as portability, ability to mimic physiological flow conditions, and reduction of the number of reagents required for preparation and detection, they lack real-time analyte detection with high accuracy. To address this limitation, biosensor technologies have been integrated with OOC systems to facilitate simultaneous analysis of different analytes with a single device. However, the integration of biosensors with OOC systems is challenging because of the competing demands of low-cost, simple fabrication processes and speed. In this study, we fabricate a glucose-sensing device and integrate it with a liver-on-a-chip (LOC) platform. A carbon black-polylactic acid-based three-electrode system was printed using fused deposit molding 3D printing technology to simplify the fabrication process. The sensitivity of the fabricated glucose biosensing device was enhanced by coating the electrodes with multi-walled carbon nanotubes. A biosensing integration study performed using a perfusion-based LOC demonstrated the stability, biocompatibility, and sensitivity of the proposed glucose sensing device. Furthermore, drug-toxicity studies conducted using the LOC platform demonstrated the ability of the device to detect a broad range of glucose concentrations and its enhanced sensitivity.


Procalcitonin and blood lactate level as predictive biomarkers in pediatric multiple trauma patients' pediatric intensive care outcomes: A retrospective observational study.
Pediatric trauma represents a significant source of morbidity and mortality in children, encompassing a broad spectrum of injuries. Despite advancements in the treatment and prevention of injuries, the risk of trauma in children remains a persistent concern. Severe trauma cases often necessitate admission to a pediatric intensive care unit (PICU). Procalcitonin, an essential biomarker that elevates bacterial infections and trauma, and elevated lactate levels can signal adverse outcomes in critically ill patients. This study retrospectively examined pediatric patients with multiple trauma treated at the Başakşehir Çam and Sakura City Hospital PICU between 2021 and 2023. The analysis sought to evaluate the relationship between initial procalcitonin and lactate levels with the duration of stay in the PICU, the length of invasive mechanical ventilation (IMV), and the duration of inotropic support. Furthermore, a comparison was made between procalcitonin and lactate levels in survivors and non-survivors, analyzing their potential influence on PICU outcomes and mortality. For pediatric multi-trauma patients, the median duration of stay in the PICU was found to be 3 days. Among these patients, 32% necessitated IMV support and utilized it for a median of 5 days. Additionally, 36% of these patients were provided inotropic drug support for a median time of 6 days. The observed mortality rate was 11%. Procalcitonin and blood lactate levels were found to have significant predictive power for mortality with odds ratios of 1.05 (P = .04) and 1.87 (P = .02), respectively. Both blood lactate and procalcitonin levels were significantly associated with the duration of IMV support, the period of inotropic drug administration, and the length of PICU stay (P < .01; P < .01; P < .01, respectively). this research underscores the prognostic value of initial procalcitonin and lactate levels about the intensive care trajectory of pediatric trauma patients. The findings suggest that both procalcitonin and lactate levels may play pivotal roles as potential biomarkers in predicting and managing clinical outcomes in this population.


Repair of upper-GI fistulas and anastomotic leakage by the use of endoluminal vacuum-assisted closure.
GI perforations, leaks, and fistulas are types of full-thickness mural defects that frequently occur as adverse events from GI surgeries such as esophagectomy for malignancy and bariatric surgery. Historically, treatment has entailed a combination of reoperation, percutaneous drainage, and bowel rest. Recently, there has been a changing paradigm in the management of these defects. Endoscopic interventions, including endoclipping and placement of self-expanding metal stents (SEMSs), have been increasingly used with good success. Despite this, some defects remain refractory to these techniques. Endoscopic vacuum-assisted closure (EVAC) is a new, promising endoscopic approach to repairing these defects. EVAC works through applying continuous, controlled negative pressure at the defect with the use of an endoscopically placed polyurethane sponge connected to an electronic vacuum device. EVAC has been shown to be feasible, safe, and effective. We present a video series of 3 cases demonstrating the successful application of EVAC for the treatment of anastomotic leakage after esophagectomy and of fistula formation after bariatric surgery. Two patients experienced anastomotic leakage after esophagectomy for esophageal adenocarcinoma, and 1 patient experienced a chronic gastric fistula after Roux-en-Y gastric bypass. The gastric bypass patient's fistula failed to resolve with over-the-scope-clip placement, and all 3 patients' defects did not heal despite SEMS placement; therefore, EVAC was performed. The bariatric surgery patient required 9 sponge exchanges over 35 days, and the 2 esophagectomy patients each required 3 sponge exchanges over 13 days. All 3 patients had resolution of their defects with EVAC. No adverse events occurred, and all patients have had no recurrence for several months. These cases help to highlight the feasibility, safety, and efficacy of EVAC for the closure of full-thickness GI defects. On the basis of our experience, the use of EVAC should be considered for these complex and refractory cases.


Sex expression and breeding strategy in Commelina benghalensis L.
This paper describes the results of a series of experiments conducted to unravel the patterns of sex expression and reproductive output in a fascinating species with high variation in sexuality. Commelina benghalensis L., an andromonoecious rainy season weed, bears male and bisexual flowers in axillary spathes of all the plants investigated. Bisexual flowers are of two types; chasmogamous (CH) and cleistogamous (CL). The former are borne on subaerial and the latter on subterranean shoots, in addition to those on aerial spathes. Three populations of the species, designated JU1, JU2 and JU3, were scanned for three consecutive years from 1996 to 1998, and the number and distribution of male, CH and CL flowers per plant were found to vary. The mere number of CH/CL flowers per plant is by itself not an accurate measure of mixed mating. It is necessary to confirm that CH flowers actually outcross and, if they do so, to what extent. Comparison of the pollen/ovule (P/O) ratio and percentage pollen germination on the stigmas of the CH and CL flowers have been used as indices of the pollination system. Confirmation of this was sought from the fruit and seed sets obtained after manual pollination of emasculated flowers with self- and cross-pollen. Results so obtained were compared with those of natural pollination. In the majority of CH flowers, the male and female reproductive phases (i.e. anther dehiscence and stigma receptivity) overlap, providing for self-pollination. However, two exceptions to this general behaviour were found in some plants of all the three populations. In some CH flowers, the female phase matures prior to anther dehiscence while in others, the anthers are sterile. Such plants, designated as variants 1 and 2, respectively, facilitate cross-pollination. While the CL flowers contribute to the production of selfed progeny, the variants of CH ones permit formation of outcrossed progeny, indicating a mixed mating strategy in C. benghalensis.


Prognostic plasma protein panel for Aβ deposition in the brain in Alzheimer's disease.
Alzheimer's disease (AD) is the most common age-associated dementia. Many studies have sought to predict cerebral amyloid deposition, the major pathological hallmark of AD, using body fluids such as blood or cerebral spinal fluid (CSF). The use of blood in diagnostic procedures is widespread in medicine; however, existing blood biomarkers for AD remain unreliable. We sought to discover blood biomarkers that discriminate Aβ deposition status in the brain. This study used 107 individuals who were cognitively normal (CN), 107 patients with mild cognitive impairment (MCI), and 40 AD patients with Pittsburg compound B positron emission tomography (PiB-PET) amyloid imaging data available. We found five plasma biomarker candidates via mass spectrometry (MS) based-proteomic analysis and validated these proteins using enzyme-linked immunosorbent assay (ELISA). Our integrated models were highly predictive of brain amyloid deposition, exhibiting 0.871 accuracy with 79% sensitivity and 84% specificity overall, and 0.836 accuracy with 68% sensitivity and 90% specificity in patients with MCI. These results indicated that a combination of proteomic-based blood proteins might be a possible biomarker set for predicting cerebral amyloid deposition.


Long-term follow-up of a cohort with post sleeve gastrectomy leaks: results of endoscopic treatment and salvage surgery.
Laparoscopic sleeve gastrectomy (LSG) is the most performed bariatric procedure worldwide. The most challenging postoperative complication is gastric leak. The objectives of this study are to examine the efficacy and morbidity of different therapeutic strategies addressing leakage, and the long-term outcomes of a cohort of LSG leaks. A retrospective review of patients treated for LSG leaks between September 2014 and January 2023 at our high-volume bariatric surgery center was performed. The charts of 37 patients (29 women and 8 men) were reviewed, with a mean age of 43 years and a median follow-up of 24 months. The mean preoperative body mass index was 45.1 kg/m2. Overall, 30/37 (81%) patients were successfully treated with endoscopic management, and 7/37 (19%) ultimately underwent salvage surgery. If the leak was diagnosed earlier than 6 weeks, endoscopic treatment had a 97% success rate. The median number of endoscopic procedures was 2 per patient, and included internal pigtails, stents, septoplasty, endoluminal vacuum therapy and over-the-scope clips. Complications included stent-related ulcers (10), esophageal stenosis requiring endoscopic dilatations (4), stent migrations (2) and kinking requiring repositioning (1), and internal pigtail migration (3). Revisional surgery consisted of proximal gastrectomy and Roux-en-Y esophago-jejunal anastomosis, Roux-en-Y fistulo-jejunostomy or classic Roux-en-Y gastric bypass proximal to the gastric stricture. In 62% of the cases, the axis/caliber of the LSG was abnormal. Beyond 4 attempts, endoscopy was unsuccessful. The success rate of endoscopic management dropped to 25% when treatment was initiated more than 45 days after the index surgery. Purely endoscopic management was successful in 81% of cases; with 97% success rate if diagnosis earlier than 6 weeks. After four failed endoscopic procedures, a surgical approach should be considered. Delayed diagnosis appears to be a significant risk factor for failure of endoscopic treatment.


Comparative analysis of chromosome-encoded microcins.
Microcins are ribosomally synthesized peptide antibiotics that are produced by enterobacterial strains. Although the first studies concentrated on plasmid-encoded activities, in the last years three chromosome-encoded microcins have been described: H47, E492, and M. Here, a new microcin, I47, is presented as a fourth member of this group. Common features exhibited by chromosome-encoded microcins were searched for. The comparison of the genetic clusters responsible for microcin production revealed a preserved general scheme. The clusters essentially comprise a pair of activity-immunity genes which determine antibiotic specificity and a set of microcin maturation and secretion genes which are invariably present and whose protein products are highly homologous among the different producing strains. A strict functional relationship between the maturation and secretion pathways of microcins H47, I47, and E492 was demonstrated through genetic analyses, which included heterologous complementation assays. The peptide precursors of these microcins share a maturation process which implies the addition of a catecholate siderophore of the salmochelin type. Microcins thus acquire the ability to enter gram-negative cells through the catechol receptors. In addition, they employ a common mode of secretion to reach the external milieu by means of a type I export apparatus. The results presented herein lead us to propose that chromosome-encoded microcins constitute a defined subgroup of peptide antibiotics which are strictly related by their modes of synthesis, secretion, and uptake.


Efflux Pump Control Alters Synthetic Gene Circuit Function.
Synthetic biology aims to design new biological systems for predefined purposes, such as the controlled secretion of biofuels, pharmaceuticals, or other chemicals. Synthetic gene circuits regulating an efflux pump from the ATP-binding cassette (ABC) protein family could achieve this. However, ABC efflux pumps can also drive out intracellular inducer molecules that control the gene circuits. This will introduce an implicit feedback that could alter gene circuit function in ways that are poorly understood. Here, we used two synthetic gene circuits inducible by tetracycline family molecules to regulate the expression of a yeast ABC pump (Pdr5p) that pumps out the inducer. Pdr5p altered the dose-responses of the original gene circuits substantially in Saccharomyces cerevisiae. While one aspect of the change could be attributed to the efflux pumping function of Pdr5p, another aspect remained unexplained. Quantitative modeling indicated that reduced regulator gene expression in addition to efflux pump function could fully explain the altered dose-responses. These predictions were validated experimentally. Overall, we highlight how efflux pumps can alter gene circuit dynamics and demonstrate the utility of mathematical modeling in understanding synthetic gene circuit function in new circumstances.


A Three-dimensional Simulation Based on Radiofrequency Electrothermal Coupling fields for Skin Rejuvenation.
Radiofrequency (RF) current is used as an effective non-ablative method for skin rejuvenation. However, mixed results have been reported using different home-use RF devices. In order to evaluate the safety and effectiveness of home-use RF devices, this study has provided a three-dimensional (3D) simulation procedure based on the electrothermal coupling model for home-use RF devices. Firstly, the tissue geometric model with the setting electrode shapes was established and then imported into the simulation software. Secondly, electrical and thermal boundary conditions with excitation voltages were loaded to the corresponding components. In addition, the items of 3D temperatures at all locations and key temperatures of the tissue were assessed. The results have shown the temperature distributions of four commercial RF products, respectively. This 3D RF electrothermal coupling simulation can be conducted quickly and effectively to obtain the temperature and electric distribution of the home-use RF devices at different using periods, which is also useful for the design of home-use RF devices.Clinical Relevance- This study provides a simple and effective simulation procedure for device developers to evaluate the home-use RF devices when designing products. This simulation is also helpful for customer decision-making and performance evaluation considering different devices.


De facto long-term opioid therapy for noncancer pain.
This paper describes characteristics of opioid use episodes for noncancer pain and defines thresholds for de facto long-term opioid therapy. CONSORT (CONsortium to Study Opioid Risks and Trends) includes adult members of 2 health plans serving over 1% of the US population. Opioid use episodes beginning in the years 1997 to 2005 were classified as acute, episodic, long-term/lower dose, or long-term/higher dose. On the basis of evaluation of the likelihood of opioid use continuing, long-term opioid therapy was defined by episodes lasting longer than 90 days with 10+ opioid prescriptions or 120+ days supply of opioids dispensed. Long-term/higher dose episodes (<1.5% of all opioid use episodes) were characterized by daily or near daily use, a mean duration of about 1000 days, and an average daily dose of about 55 mg. They accounted for more than half the total morphine equivalents dispensed from the years 1997 to 2006. Short-acting, non-Schedule II opioids (eg, hydrocodone with acetaminophen) were, by far, the most commonly prescribed medications for acute, episodic, and long-term episodes. Long-acting (sustained-release) opioids were the predominately prescribed medication in a minority of long-term episodes (6% to 12%). Long-term opioid therapy was characterized by the diversity in medications prescribed, dosage levels, and frequency of use. The proposed threshold for long-term opioid therapy provides a checkpoint for physicians to review whether an explicit decision to sustain opioid therapy has been reached, and to ensure that a documented treatment plan and provisions for monitoring medication use and patient outcomes are in place.


In vitro and in vivo studies of a bone morphogenetic protein-2 expressing adenoviral vector.
Bone morphogenetic proteins (BMPs) play important roles in the migration of osteoblast progenitor cells, the proliferation of mesenchymal cells, and their differentiation into chondrogenic and osteogenic cells. However, the optimum procedure to deliver BMPs remains unknown. To examine the effectiveness of a gene transfer procedure for the delivery of BMP-2, we constructed a human BMP-2-expressing replication-deficient adenoviral vector, AxCAOBMP-2, and evaluated its osteoinductive activity in vitro and in vivo. C2C12 myoblasts were infected in vitro with this viral vector or an Escherichia coli LacZ gene-expressing control adenovirus vector (AxCALacZ). Twenty-four hours after the infection, indirect immunofluorescence was performed. On day 5 after the infection, alkaline phosphatase (ALP) in the cells and osteocalcin in the culture medium were measured. Furthermore, to examine the effectiveness of gene transfer of BMP-2 in vivo, we evaluated osteoinduction by AxCAOBMP-2, under transient immunosuppression with cyclophosphamide, given at a dose of 125 mg/kg intraperitoneally the day before injection of the adenoviral vector. Twenty-five microliters of AxCAOBMP-2 (8.75 x 10(8) plaque-forming units [pfu], Group I) and AxCALacZ (1.75 x 10(8) pfu, control group) and 5 microl of AxCAOBMP-2 (1.75 x 10(8) pfu, Group II) were injected into a right calf muscle of Wistar rats. On day 21, bone formation in each group was investigated radiologically and histologically. Abundant BMP-2 expression in C2C12 cells infected with this viral vector was confirmed by immunofluorescence. C2C12 cells transferred with the BMP-2 gene by this vector produced ALP in the cells and also produced and secreted osteocalcin in the culture medium. Osteoinduction was found only in the AxCAOBMP-2 treated groups with immunosuppression. Osteoinduction activity was higher in Group I than in Group II. This study demonstrated the osteoinductive activity in vitro and in vivo by an adenoviral vector carrying the BMP-2 gene. Gene therapy with AxCAOBMP-2 under transient immunosuppression may be useful for bone reconstruction.


Do interactions between stress and immune responses lead to symptom exacerbations in irritable bowel syndrome?
Irritable bowel syndrome (IBS) is a common, debilitating gastrointestinal (GI) disorder, with a worldwide prevalence of between 10% and 20%. This functional gut disorder is characterized by episodic exacerbations of a cluster of symptoms including abdominal pain, bloating and altered bowel habit, including diarrhea and/or constipation. Risk factors for the development of IBS include a family history of the disorder, childhood trauma and prior gastrointestinal infection. It is generally accepted that brain-gut axis dysfunction is fundamental to the development of IBS; however the underlying pathophysiological mechanisms remain elusive. Additional considerations in comprehending the chronic relapsing pattern that typifies IBS symptoms are the effects of both psychosocial and infection-related stresses. Indeed, co-morbidity with mood disorders such as depression and anxiety is common in IBS. Accumulating evidence points to a role for a maladaptive stress response in the initiation, persistence and severity of IBS-associated symptom flare-ups. Moreover, mechanistically, the stress-induced secretion of corticotropin-releasing factor (CRF) is known to mediate changes in GI function. Activation of the immune system also appears to be important in the generation of IBS symptoms and increasing evidence now implicates low-grade inflammation or immune activation in IBS pathophysiology. There is a growing body of research focused on understanding at a molecular, cellular and in vivo level, the relationship between the dysregulated stress response and immune system alterations (either individually or in combination) in the etiology of IBS and to the occurrence of symptoms.


A novel antioxidant multitarget iron chelator M30 protects hepatocytes against ethanol-induced injury.
The multitarget iron chelator, M30, is a novel antioxidant and protective agent against oxidative stress in a spectrum of diseases. However, there is no report regarding its role in liver diseases. Since oxidative stress is one of the major pathological events during the progression of alcoholic liver diseases, the protective effects and mechanisms of M30 on ethanol-induced hepatocyte injury were investigated in this study. Rat hepatocyte line BRL-3A was pretreated with M30 prior to ethanol treatment. Cell death, apoptosis, oxidative stress, and inflammation were examined. Specific antagonists and agonists were applied to determine the involvements of hypoxia inducible factor-1 alpha (HIF-1α) and its upstream adenylate cyclase (AC)/cyclic AMP (cAMP)/protein kinase A (PKA)/HIF-1α/NOD-like receptor 3 (NLRP3) inflammasome pathway. We found that M30 significantly attenuated ethanol-induced cellular death, apoptosis, production of reactive oxygen species (ROS), and secretion of inflammatory cytokines and inhibited activation of the AC/cAMP/PKA/HIF-1α/NLRP3 inflammasome pathway. Inhibition and activation of the AC/cAMP/PKA/HIF-1α pathway mimicked and abolished the effects of M30, respectively. In conclusion, inhibition of the AC/cAMP/PKA/HIF-1α/NLRP3 inflammasome pathway by M30 partially contributes to its attenuation of hepatocyte injury caused by ethanol exposure.


Characterization of the interaction of lambda exonuclease with the ends of DNA.
Lambda exonuclease processively degrades one strand of double-stranded DNA (dsDNA) in the 5"-3" direction. To understand the mechanism through which this enzyme generates high processivity we are analyzing the first step in the reaction, namely the interaction of lambda exonuclease with the ends of substrate DNA. Endonuclease mapping of lambda exonuclease bound to DNA has shown that the enzyme protects approximately 13-14 bp on dsDNA, and no nucleo-tides on the single-stranded tail of the DNA product. We have developed a rapid fluorescence-based assay using 2-aminopurine and measured the steady-state rate constants for different end-structures of DNA. The relative k(cat)for 5" ends decreases in the order 5" recessed > blunt >> 5" overhang. However, k(cat)/K(m)remains relatively constant for these different structures suggesting they are all used equally efficiently as substrates. From these data we propose that a single-stranded 5" overhang end can bind non-productively to the enzyme and the non-hydrolyzed strand is required to aid in the proper alignment of the 5" end. We have also measured the length-dependence of the steady-state rate para-meters and find that they are consistent with a high degree of processivity.


Exuberant cellular reaction of the optic nerves in experimental Creutzfeldt-Jakob disease.
We report here on the exuberant glial reaction in the optic nerves affected by prion diseases. Optic nerves from CJD- and GSS-, and scrapie-infected mice and hamsters showed severe pathology. These lesions were qualitatively indistinguishable from each other but were more intense in the Fujisaki model than in the hamsters inoculated with Echigo-1. Exuberant cellular reaction comprised of macrophages containing numerous mitochondria, abundant rough endoplasmic reticulum, and secondary lysosomes filled with digested myelin debris, electron-dense material and occasionally, entire myelin-bound vacuoles were readily observed in both models. Macrophages actively digesting myelin fragments and containing lyre-like bodies and paracrystalline inclusions were frequently noted. Some macrophages extended long filopodia to form labyrinth-like structures, and within a few macrophages, concentric arrays of cisterns and channels sequestrated part of the cytoplasm. An analogous network of narrow cisterns was seen to surround whole segments of the myelinated fibers.


Development and validation of an instrument to measure the effects of a mistletoe preparation on quality of life of cancer patients: the Life Quality Lectin-53 (LQL-53) Questionnaire.
Mistletoe preparations standardised to the content of mistletoe lectin are supposed to improve quality of life (QoL) in patients with cancer. To obtain a validated and sensitive research instrument, the Life Quality Lectin-53 (LQL-53) Questionnaire was developed in three phases: item generation via interviews with 42 patients, item selection by means of a study with 109 cancer patients, and psychometric testing. The LQL-53 includes 46 items assigned to the subscales 'General well-being', 'Emotional well-being', 'Vitality' 'Hope', 'Locus of control', Social relationships', and 'Physical complaints', plus seven items dealing with possible adverse effects of mistletoe treatment. Psychometric testing was carried out in a study with 112 patients with solid tumours who were treated with a mistletoe preparation standardised to mistletoe lectins for 12 weeks. Internal consistency (Cronbach's alpha) was between 0.72 and 0.94. Test-retest reliability was > or = r of 0.72. Subscales correlated highly with external criteria. Construct validity, as determined by Multitrait Scaling Analysis, resulted in an optimal assignment of items to subscales (scaling success) for four of the subscales. During the course of the therapy, significant improvement in QoL was found in all subscales. In two of the subscales, effect size was high (>0.80), and in five other subscales it varied between 0.53 and 0.78.


Dietary supplementation with 1-kestose induces altered locomotor activity and increased striatal dopamine levels with a change in gut microbiota in male mice.
1-Kestose (KES), a dietary fiber and prebiotic carbohydrate, benefits various physiological functions. This study aimed to examine whether diets supplemented with KES over three consecutive generations could significantly affect some host physiological aspects, including behavioral phenotypes and gut microbial ecology. Mice that received KES-supplemented diets for three generations demonstrated increased activity compared with those fed diets lacking KES. Furthermore, the KES group showed increased striatal dopamine (DA) and serotonin (5-HT) levels. The observed increase in DA levels within the striatum was positively correlated with locomotor activity in the KES group but not in the control (CON) group. The α-diversities were significantly lower in the KES group compared to the CON group. The three-dimensional principal coordinate analysis revealed a substantial distinction between the KES and CON groups across each generation. At the genus level, most gut microbiota genera exhibited lower abundances in the KES group than in the CON group, except for Bifidobacteria and Akkermansia. Spearman's rank-order analysis indicated significant negative correlations between the striatal DA levels and α-diversity values. These findings suggest that prolonged supplementation with KES may stimulate increased locomotor activity along with elevated striatal DA levels, which are potentially associated with KES-induced alterations in the gut microbiota.


COVID-19 Infection: Concise Review Based on the Immunological Perspective.
The outbreak of coronavirus disease 2019 (COVID-19) has posed a serious threat to public health. There is an urgent need for discovery methods for the prevention and treatment of COVID-19 infection. Understanding immunogenicity together with immune responses are expected to provide further information about this virus. We hope that this narrative review article may create new insights for researchers to take great strides toward designing vaccines and novel therapies in the near future. The functional properties of the immune system in COVID-19 infection is not exactly clarified yet. This is compounded by the many gaps in our understanding of the SARS-CoV-2 immunogenicity properties. Possible immune responses according to current literature are discussed as the first line of defense and acquired immunity. Here, we focus on proposed modern preventive immunotherapy methods in COVID-19 infection.


[Studies on the virulent factor of Escherichia coli isolated from urogenital infection--pilus type and adherence to human exfoliated uroepithelial cells etc].
One hundred and twenty three strains of Escherichia coli isolated from the urine of patients with urogenital infections and 55 strains isolated from the fecal samples of healthy individuals were assayed for 0 antigen and hemolysin production as virulence factors, and for pilus type and in vitro tests of adhesion to human exfoliated uroepithelial cells as colonization factors. The incidence of MS pili in Escherichia coli isolated from patients with acute uncomplicated cystitis, chronic complicated cystitis, acute uncomplicated pyelonephritis and acute prostatitis was 60.9%, 22.7%, 87.5%, 68.2% and 30.9%, respectively. The incidence of P pili was 69.7%, 51.2%, 62.5%, 54.5% and 25.5%, respectively. The strains showing haemolysin production had MS pili and P pili, which show strong virulence. MS pili strains and P pili strains isolated from the patients with acute uncomplicated cystitis and acute uncomplicated pyelonephritis adhered to human exfoliated uroepithelial cells well. Consequently, the pilus type might be the most significant colonization factor in uncomplicated urogenital infection which is shown by the normal defense mechanism in host side.


Cumulate xenolith in oahu, hawaii: implications for deep magma chambers and hawaiian volcanism.
The maximum depth at which large (>1000 km(3)) terrestrial mafic magma chambers can form has generally been thought to be the Moho, which occurs at a mean depth of about 35 kilometers beneath the continents and 8 kilometers beneath ocean basins. However, the presence of layers of cumulus magnesium-rich spinel and olivine and intercumulus garnet in an unusual mantle xenolith from Oahu, Hawaii, suggests that this rock is a fragment of a large magma chamber that formed at a depth of about 90 kilometers; Hawaiian shield-building magmas may pond and fractionate in such magma chambers before continuing their ascent. This depth is at or near the base of the 90-million-year-old lithosphere beneath Oahu; thus, rejuvenated stage alkalic magmas containing mantle xenoliths evidently also originate below the lithosphere.


Organisational intervention to reduce occupational stress and turnover in hospital nurses in the Northern Territory, Australia.
To evaluate the impact of an organisational intervention aimed to reduce occupational stress and turnover rates of 55% in hospital nurses. The evaluation used a pre- and post-intervention design, triangulating data from surveys and archival information. Two public hospitals (H1 and H2) in the Northern Territory (NT) Australia participated in the intervention. 484 nurses from the two NT hospitals (H1, Wave 1, N = 103, Wave 2, N = 173; H2, Wave 1, N = 75, Wave 2, N = 133) responded to questionnaires administered in 2008 and in 2010. The intervention included strategies such as the development and implementation of a nursing workload tool to assess nurse workloads, roster audits, increased numbers of nursing personnel to address shortfall, increased access to clinical supervision and support for graduates, increased access to professional development including postgraduate and short courses, and a recruitment campaign for new graduates and continuing employees. We used an extended Job Demand-Resources framework to evaluate the intervention and 17 evaluation indicators canvassing psychological distress, emotional exhaustion, work engagement, job satisfaction, job demands, job resources, and system factors such as psychosocial safety climate. Turnover rates were obtained from archival data. Results demonstrated a significant reduction in psychological distress and emotional exhaustion and a significant improvement in job satisfaction, across both hospitals, and a reduction in turnover in H2 from 2008 and 2010. Evidence suggests that the intervention led to significant improvements in system capacity (adaptability, communication) in combination with a reduction in job demands in both hospitals, and an increase in resources (supervisor and coworker support, and job control) particularly in H1. The research addresses a gap in the theoretical and intervention literature regarding system/organisation level approaches to occupational stress. The approach was very successful on a range of health, work outcome, and job design indicators with results providing compelling evidence for the success of the system/organisational level intervention. The quasi-experimental design enabled us to conclude that improvements for the nurses and midwives could be attributed to the organisational intervention by the NT Department of Health (DoH). Further research should be undertaken to explore longer-term impacts, and particularly the influence on turnover. Levels of stress in hospital nurses remain high and present important implications for the psychological well-being of staff.


Assessing Fatty Infiltration of Paraspinal Muscles in Patients With Lumbar Spinal Stenosis: Goutallier Classification and Quantitative MRI Measurements.
Objective: Fatty infiltration of paraspinal muscle is associated with spinal disorders. It can be assessed qualitatively (i.e., Goutallier classification) and quantitatively using image processing software. The aims of this study were to compare paraspinal muscle fatty infiltration as assessed using the Goutallier classification vs. quantitative magnetic resonance images (MRI) measurements and to investigate the association between anthropometric parameters and paraspinal muscle morphology and fatty infiltration in patients with symptomatic lumbar spinal stenosis (LSS). Methods: Patients affected by symptomatic LSS scheduled for surgery with available MRI of the lumbar spine were included in this retrospective cross-sectional study. Fatty infiltration at each lumbar level was rated qualitatively according to the Goutallier classification and quantified based on the cross-sectional area (CSA) of the paraspinal muscle, of its lean fraction (LeanCSA), and the ratio between LeanCSA and CSA and the CSA relative to the CSA of vertebral body (RCSA). Considering the muscle as a single unit, overall fatty infiltration according to Goutallier, overall CSA, LeanCSA, LeanCSA/CSA, and RCSA were computed as averages (aGoutallier, aCSA, aLeanCSA, aLeanCSA/aCSA, and aRCSA). Associations among parameters were assessed using Spearman's respective Pearson's correlation coefficients. Results: Eighteen patients, with a mean age of 71.3 years, were included. aGoutallier correlated strongly with aLeanCSA and aLeanCSA/aCSA (R = -0.673 and R = -0.754, both P < 0.001). There was a very strong correlation between values of the left and right sides for CSA (R = 0.956, P < 0.001), LeanCSA (R = 0.900, P < 0.001), and LeanCSA/CSA (R = 0.827, P < 0.001) at all levels. Among all anthropometric measurements, paraspinal muscle CSA correlated the most with height (left: R = 0.737, P < 0.001; right: R = 0.700, P < 0.001), while there was a moderate correlation between vertebral body CSA and paraspinal muscle CSA (left: R = 0.448, P < 0.001; right: R = 0.454, P < 0.001). Paraspinal muscle CSA correlated moderately with body mass index (BMI; left: R = 0.423, P < 0.001; right: R = 0.436, P < 0.001), and there was no significant correlation between aLeanCSA or aLeanCSA/CSA and BMI. Conclusions: The Goutallier classification is a reliable yet efficient tool for assessing fatty infiltration of paraspinal muscles in patients with symptomatic LSS. We suggest taking body height as a reference for normalization in future studies assessing paraspinal muscle atrophy and fatty infiltration.


Functional Transcranial Doppler Ultrasound for Measurement of Hemispheric Lateralization During Visual Memory and Visual Search Cognitive Tasks.
Functional transcranial Doppler ultrasound (fTCD) is a noninvasive sensing modality that measures cerebral blood flow velocity (CBFV) with high temporal resolution. CBFV change is correlated to changes in cerebral oxygen uptake, enabling fTCD to measure brain activity and lateralization with high accuracy. However, few studies have examined the relationship of CBFV change during visual search and visual memory tasks. Here a protocol to compare lateralization between these two similar cognitive tasks using fTCD is demonstrated. Ten healthy volunteers (age 21±2 years) were shown visual scenes on a computer and performed visual search and visual memory tasks while CBFV in the bilateral middle cerebral arteries was monitored with fTCD. Each subject completed 40 trials, consisting of baseline (25 s), calibration (variable), instruction (2.5 s), and task (20 s) epochs. Lateralization was computed for each task by calculating the bilateral CBFV envelope percent change from baseline and subtracting the right side from the left side. The results showed significant lateralization ( ) of the visual memory and visual search tasks, with memory reaching lateralization of 1.6% and search reaching lateralization of 0.5%, suggesting that search is more right lateralized (and therefore may be related to "holistic" or global perception) and memory is more left lateralized (and therefore may be related to local perception). This method could be used to compare cerebral activity for any related cognitive tasks as long as the same stimulus is used in all tasks. The protocol is straightforward and the equipment is inexpensive, introducing a low-cost high temporal resolution technique to further study lateralization of the brain.


Puerarin protects rat brain against ischemia/reperfusion injury by suppressing autophagy via the AMPK-mTOR-ULK1 signaling pathway.
Puerarin suppresses autophagy to alleviate cerebral ischemia/reperfusion injury, and accumulating evidence indicates that the AMPK-mTOR signaling pathway regulates the activation of the autophagy pathway through the coordinated phosphorylation of ULK1. In this study, we investigated the mechanisms underlying the neuroprotective effect of puerarin and its role in modulating autophagy via the AMPK-mTOR-ULK1 signaling pathway in the rat middle cerebral artery occlusion model of cerebral ischemia/reperfusion injury. Rats were intraperitoneally injected with puerarin, 50 or 100 mg/kg, daily for 7 days. Then, 30 minutes after the final administration, rats were subjected to transient middle cerebral artery occlusion for 90 minutes. Then, after 24 hours of reperfusion, the Longa score and infarct volume were evaluated in each group. Autophagosome formation was observed by transmission electron microscopy. LC3, Beclin-1 p62, AMPK, mTOR and ULK1 protein expression levels were examined by immunofluorescence and western blot assay. Puerarin substantially reduced the Longa score and infarct volume, and it lessened autophagosome formation in the hippocampal CA1 area following cerebral ischemia/reperfusion injury in a dose-dependent manner. Pretreatment with puerarin (50 or 100 mg/kg) reduced Beclin-1 expression and the LC3-II/LC3-I ratio, as well as p-AMPK and pS317-ULK1 levels. In comparison, it increased p62 expression. Furthermore, puerarin at 100 mg/kg dramatically increased the levels of p-mTOR and pS757-ULK1 in the hippocampus on the ischemic side. Our findings suggest that puerarin alleviates autophagy by activating the APMK-mTOR-ULK1 signaling pathway. Thus, puerarin might have therapeutic potential for treating cerebral ischemia/reperfusion injury.


Pressure-independent effects of AT1-receptor antagonism on cardiovascular remodeling in aortic-banded rats.
To determine the role of angiotensin II-receptor blockade on cardiovascular remodeling in a pressure-overload model of cardiac hypertrophy, a subdiaphragmatic aortic band was placed in adult male Sprague-Dawley rats. Aortic-banded (AB) rats were left untreated or were losartan (Los; 250 mg/l) treated (AB-Los). Sham-operated (S) controls were either left untreated or treated with Los (S-Los). After 4 wk, rats were catheterized for measurement of mean arterial pressures [carotid (CMAP) and femoral (FMAP), in mmHg]. Hearts were perfused on a modified Langendorff system, and minimal coronary resistance (MCR) was determined. Hearts were then perfusion fixed, total and regional heart weights were recorded, and sections were processed for morphology. Changes in coronary artery medial thickness and perivascular fibrosis were assessed by quantitative image analysis. CMAP was significantly higher in AB and AB-Los than in S or S-Los (P < 0.05). There was no difference in FMAP in AB vs. S, but AB-Los and S-Los had lower FMAPs than S. Total heart weight and left ventricular weight-to-body weight ratios were increased in AB and AB-Los compared with S and S-Los (P < 0.05). MCR of AB was greater than S and S-Los. MCR of AB-Los was significantly lower than AB and was not significantly different from S and S-Los. In coronary vessels, medial thickness was greatest in AB, whereas there was no difference among AB-Los, S, and S-Los. Similarly, the increase in perivascular fibrosis was greatest in AB, and there was no difference among AB-Los, S, and S-Los. These data suggest that angiotensin II, independent of increased arterial pressure, is critical for the development of the vascular and fibrotic changes that occur in this model of pressure-overload hypertrophy.


Orbital osteoma: three case reports.
Osteoma is the most frequent benign tumor of paranasal sinuses. It is generally asymptomatic and usually discovered by chance during radiological imaging. Three cases with orbital extension are reported and discussed. 1st observation: Mrs. M.N. is an 18 years-old caucasian female who presented a stony orbital deformity associated with chronical dacryocystitis. Tomography of the orbit revealed a probable ethmoidal osteoma compressing the lacrimal canals. The management consisted in a surgical excision of the whole tumor by external approach, associated with a dacryocystorhinostomy. The patient's follow-up for the last 12 months was normal. 2nd observation: Miss K.A. is a 16 years-old caucasian female who came to consultation for a swelling of the medial angle of the left eye. Computed tomography images showed a fronto-ethmoidal process displaying a bone density consistent with an osteoma. The tumor was removed through an external ethmoidectomy. The patient was free of symptoms at 6 months follow-up. 3rd observation: Mrs. F. Z. is a 45 years-old patient who presented a 5 years history of right painful exophthalmos. The computed tomography was consistent with a fronto-ethmoidal osteoma with intraorbital extension near the optic nerve. The surgical excision was limited to the intraorbital portion. Postoperative complications included ptosis and diplopia. Osteomas most commonly affect the fronto-ethmoid sinuses. They rarely show intraorbital extension or cause intracranial complications. They are generally asymptomatic. Symptoms are generally of late onset and are a consequence of tumoral growth and compression of neighbouring organs, as it is the case in our patients. Tumoral exophtalmos is the major ophthalmological sign. The computed tomography is of a great contribution, not only for diagnosis but also for the choice of the surgical approach. The classical surgical technique consists generally in a surgical excision of the osteoma. This surgery may induce ocular or neurochirurgical complications. Osteomas of the paranasal sinuses are usually asymptomatic. If they become voluminous, they may cause orbital manifestations and serious complications. The rarity of ethmoidal osteoma with orbital growth made our cases interesting to report.


Profiling the regulatory lipids: another systemic way to unveil the biological mystery.
The regulatory lipids are a class of bioactive lipids which regulate various important biological processes. Profiling these regulatory lipids is an attractive method to understand the role of these metabolites. This is especially true because most of these regulatory lipids are derived from several important pharmacological targets: cyclooxygenase, lipoxygenase, and cytochrome P450 enzymes. This review highlights the development of methods to profile these regulatory lipids and the recent publications employing these profiling methods. The recent development of methods for the profiling of regulatory lipids target two different directions: to expand coverage for discovery studies (fingerprinting) and to make the quantitative method more accurate, sensitive, and faster for diagnostic or more detailed studies. Recent applications of these profiling methods including assessment of in-vivo drug engagement, pathways crosstalk, and possible mechanisms for side-effects of a withdrawn anti-inflammatory drug rofecoxib are also reviewed here. The profiling of regulatory lipids is a useful tool for many investigations. The breadth of coverage, throughput limits with detection, and reproducibility of quantitation are being improved. The resulting data will assist with fundamental investigation, disease biomarker discovery, drug discovery, and drug development.


Case report: dramatic response to lamivudine therapy following corticosteroid priming in chronic hepatitis B.
A 21 year-old male patient with chronic hepatitis B was treated with lamivudine 150 mg daily after withdrawal of a short course of oral prednisolone (30 mg daily for 3 weeks, 15 mg daily for 1 week). Serum hepatitis B virus (HBV)-DNA increased during prednisolone pretherapy and serum alanine aminotransferase (ALT) was increasing after withdrawal of prednisolone. Clearance of HBV-DNA with hepatitis B e antigen seroconversion and ALT normalization occurred within 2 months after starting lamivudine therapy. If this dramatic response to lamivudine therapy after corticosteroid priming is confirmed by further studies, the regimens used in this particular case might become a powerful therapeutic tool for chronic HBV infection.


High prevalence of fecal carriage of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in a pediatric unit in Madagascar.
Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have spread worldwide but there are few reports on carriage in hospitals in low-income countries. ESBL-producing Enterobacteriaceae (ESBL-PE) have been increasingly isolated from nosocomial infections in Antananarivo, Madagascar. we conducted a prevalence survey in a pediatric unit from March to April 2008 Patient rectal swabs were sampled on the first and the last day of hospitalization. Medical staff and environment were also sampled. Rectal and environmental swabs were immediately plated onto Drigalski agar supplemented with 3 mg/liter of ceftriaxon. Fecal carriage was detected in 21.2% of 244 infants on admission and 57.1% of 154 on discharge, after more than 48 hours of hospitalization (p < 0.001). The species most frequently detected on admission were Escherichia coli and Klebsiella pneumoniae (36.9%), whereas, on discharge, K. pneumoniae was the species most frequently detected (52.7%). ESBL-associated resistances were related to trimethoprim-sulfamethoxazole (91.3%), gentamicin (76.1%), ciprofloxacin (50.0%), but not to amikacin and imipenem. The increased prevalence of carriage during hospitalization was related to standard antimicrobial therapy. The significant emergence of multidrug-resistant enteric pathogens in Malagasy hospitals poses a serious health threat requiring the implementation of surveillance and control measures for nosocomial infections.


The treatment of systemic lupus erythematosus (SLE) with the T-cell immunostimulant drug levamisole: A case report.
A 23-year-old woman with a nine-month history of SLE, uncontrolled with high daily doses of steroids, was treated with the T-cell immunostimulant drug, levamisole. Following a single oral three-day course of therapy with this agent her immunologic parameters and clinical manifestations underwent marked regression. Continuous two-weekly course of this drug at three days per course have allowed rapid reduction of her steroid and resulted in complete remission of her symptoms. The theoretical considerations for the use of levamisole dovetailed perfectly with the observed response to this agent.


Karyotype complements the International Prognostic Scoring System for primary myelofibrosis.
The International Prognostic Scoring System (IPSS) for primary myelofibrosis (PMF) is based on five independent predictors of inferior survival: age >65 yr, hemoglobin <10 g/dL, leukocyte count >25 x 10(9)/L, circulating blasts > or =1%, and presence of constitutional symptoms. The presence of 0, 1, 2, and > or =3 adverse factors defines low, intermediate-1, intermediate-2, and high risk disease, respectively. We examined the additional prognostic relevance of karyotype. World Health Organization criteria were used for PMF diagnosis. Only patients with bone marrow cytogenetic studies at the time or within 1 yr of diagnosis and a minimum of 20 evaluable metaphases were considered. Cytogenetic findings were categorized as 'normal' vs. 'abnormal' or 'favorable' (normal or with sole abnormalities of 13q- or 20q-) vs. 'unfavorable' (all other abnormalities). A total of 109 patients were studied (median age 63 yr). Numbers of patients in the above-listed four IPSS risk groups were 26, 31, 28, and 24, respectively. Cytogenetic results were abnormal in 33% of the patients and unfavorable in 21%. At a median follow-up of 35 months, 45 (41%) deaths were recorded. 'Unfavorable' (P = 0.008) but not 'abnormal' (P = 0.19) karyotype predicted shortened survival and its significance remained on multivariable analysis that included the IPSS or other prognostic tools as covariates. JAK2V617F, detected in 63 (58%) patients, was inconsequential to survival. In PMF, specific cytogenetic abnormalities and not the mere presence of an abnormal karyotype provide important prognostic information that is not accounted for by the IPSS or other established risk factors.


APOE and the Association of Fatty Acids With the Risk of Stroke, Coronary Heart Disease, and Mortality.
Background and Purpose- The role of dietary fat on cardiovascular health and mortality remains under debate. Because the APOE is central to the transport and metabolism of lipids, we examined associations between plasma fatty acids and the risk of stroke, coronary heart disease, and mortality by APOE-ε4 genotype. Methods- We included 943 FHS (Framingham Heart Study) and 1406 3C (Three-City) Bordeaux Study participants. Plasma docosahexaenoic, linoleic, arachidonic, and palmitic fatty acids were measured at baseline by gas chromatography. All-cause stroke, ischemic stroke, coronary heart disease, and all-cause mortality events were identified prospectively using standardized protocols. Each cohort used Cox models to separately relate fatty acid levels to the risk of developing each event during ≤10 years of follow-up adjusting for potential confounders and stratifying by APOE genotype (ε4 carriers versus noncarriers). We then meta-analyzed summary statistics using random-effects models. Results- On average, participants had a mean age of 74 years, 61% were women, and 21% (n=483) were APOE-ε4 carriers. Meta-analysis results showed that, only among APOE-ε4 carriers, every SD unit increase in linoleic acid was associated with a reduced risk of all-cause stroke (hazard ratio [HR], 0.54 [95% CI, 0.38-0.78]), ischemic stroke (HR, 0.48 [95% CI, 0.33-0.71]), and all-cause mortality (HR, 0.70 [95% CI, 0.57-0.85]). In contrast, every SD unit increase in palmitic acid was related to an increased risk of all-cause stroke (HR, 1.58 [95% CI, 1.16-2.17]), ischemic stroke (HR, 1.76 [95% CI, 1.26-2.45]), and coronary heart disease (HR, 1.48 [95% CI, 1.09-2.01]), also in APOE-ε4 carriers only. Results for docosahexaenoic acid and arachidonic acid were heterogeneous between cohorts. Conclusions- These exploratory results suggest that APOE-ε4 carriers may be more susceptible to the beneficial or adverse impact of fatty acids on cardiovascular disease and mortality. In this subgroup, higher linoleic acid was protective for stroke and mortality, whereas palmitic acid was a risk factor for stroke and coronary heart disease. The mechanisms underlying these novel findings warrant further investigation.


Validation of a home environment test battery for supporting assessments in advanced Parkinson's disease.
Test sequences in a test battery for Parkinson's disease patients, consisting of self-assessments and motor tests, were carried out repeatedly in a telemedicine setting, during week-long test periods and results were summarized in an 'overall score'. 35 patients in stable and fluctuating conditions (15 age- and gender-matched pairs) used the test battery for 1 week, and were then assessed with UPDRS and PDQ-39. This procedure was repeated 1 week later, without treatment changes. Reliability was assessed by intraclass correlation coefficients and Cronbach's alpha. Convergent validity was assessed by Spearman rank correlations and known-groups' validity, by the Mann-Whitney test. According to anonymous usability questionnaires, the patients could easily complete the tasks. Median compliance (93%) and test-retest reliability (0.88) were good. The correlations between overall score and total UPDRS (-0.64) and PDQ-39 (-0.72) were adequate. Median overall score was 18% better in the stable compared to the fluctuating group (p = 0.0014).


Myeloablative vs reduced intensity T-cell-replete haploidentical transplantation for hematologic malignancy.
In the absence of prospective studies that examine the effect of conditioning regimen intensity after T-cell-replete haploidentical transplant for acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), and myelodysplastic syndrome (MDS), a retrospective cohort analysis was performed. Of the 1325 eligible patients (AML, n = 818; ALL, n = 286; and MDS, n = 221), 526 patients received a myeloablative regimen and 799 received a reduced-intensity regimen. Graft-versus-host disease prophylaxis was uniform with posttransplant cyclophosphamide, a calcineurin inhibitor, and mycophenolate mofetil. The primary end point was disease-free survival. Cox regression models were built to study the effect of conditioning regimen intensity on transplant outcomes. For patients aged 18 to 54 years, disease-free survival was lower (hazard ratio [HR], 1.34; 42% vs 51%; P = .007) and relapse was higher (HR, 1.51; 44% vs 33%; P = .001) with a reduced-intensity regimen compared with a myeloablative regimen. Nonrelapse mortality did not differ according to regimen intensity. For patients aged 55 to 70 years, disease-free survival (HR, 0.97; 37% vs 43%; P = .83) and relapse (HR, 1.32; 42% vs 31%; P = .11) did not differ according to regimen intensity. Nonrelapse mortality was lower with reduced-intensity regimens (HR, 0.64; 20% vs 31%; P = .02). Myeloablative regimens are preferred for AML, ALL, and MDS; reduced-intensity regimens should be reserved for those unable to tolerate myeloablation.


High catalytic efficiency and resistance to denaturing in bacterial Rho GTPase-activating proteins.
Several major bacterial pathogens use the type III secretion system (TTSS) to deliver virulence factors into host cells. Bacterial Rho GTPase activating proteins (RhoGAPs) comprise a remarkable family of type III secreted toxins that modulate cytoskeletal dynamics and manipulate cellular signaling pathways. We show that the RhoGAP activity of Salmonella SptP and Pseudomonas ExoS toxins is resistant to variations in the concentration of NaCl or MgCl(2), unlike the known salt dependant nature of the activity of some eukaryotic GAPs such as p190, RanGAP and p120GAP. Furthermore, SptP-GAP and ExoS-GAP display full activity after treatment at 80°C or with 6 m urea, which suggests that these protein domains are capable of spontaneous folding into an active state following denaturing such as what might occur upon transit through the TTSS needle. We determined the catalytic activity of bacterial GAPs for Rac1, CDC42 and RhoA GTPases and found that ExoS, in addition to Yersinia YopE and Aeromonas AexT toxins, display higher catalytic efficiencies for Rac1 and CDC42 than the known eukaryotic GAPs, making them the most catalytically efficient RhoGAPs known. This study expands our knowledge of the mechanism of action of GAPs and of the ways bacteria mimic host activities and promote catalysis of eukaryotic signaling proteins.


Osteopontin: a component of matrix in capsular opacification and subcapsular cataract.
To examine whether tissues of human capsular opacification and subcapsular cataract contain osteopontin, an adhesive matrix protein, and whether mouse lens epithelium expresses osteopontin after injury. An immunohistochemical examination was conducted to determine whether matrices in human postoperative capsular specimens and anterior subcapsular cataract contain osteopontin. The spatial and temporal protein expression patterns of osteopontin were then determined in epithelium of a healing mouse lens after a capsular incision. Human lens epithelial cells in the specimens extracted at the time of vitrectomy 10 days after cataract surgery and also after longer healing intervals were labeled with an anti-osteopontin antibody, whereas uninjured lens epithelium was not. In the later healing phase, matrix of capsular opacification was positive for osteopontin. Lens cells amid anterior subcapsular cataract tissue were also positive. Osteopontin was detected in the cell surface and membrane and the cytoplasm of lens cells, as well as in the matrix. Unlike normal uninjured specimens, anterior lens capsule of some of the healing postoperative specimens and anterior subcapsular cataract specimens also faintly or weakly stained for osteopontin. Mouse lens epithelium started to express osteopontin protein at 8 hours after injury, before the cells changed their shape from epithelial cell type to fibroblast type. Expression of osteopontin lasted during the healing interval, even after the cells transformed into fibroblast-like cells. Extracellular matrix in human postoperative capsular opacification and anterior subcapsular cataract contains osteopontin. Epithelial cells of a mouse lens also ectopically express osteopontin in response to capsular injury.


The permeability of endplate channels to monovalent and divalent metal cations.
The relative permeability of endplate channels to monovalent and divalent metal ions was determined from reversal potentials. Thallium is the most permeant ion with a permeability ratio relative to Na+ of 2.5. The selectivity among alkali metals is weak with a sequence, Cs+ greater than Rb+ greater than K+ greater than Na+ greater than Li+, and permeability ratios of 1.4, 1.3, 1.1, 1.0, and 0.9. The selectivity among divalent ions is also weak, with a sequence for alkaline earths of Mg++ greater than Ca++ greater than Ba++ greater than Sr++. The transition metal ions Mn++, Co++, Ni++, Zn++, and Cd++ are also permeant. Permeability ratios for divalent ions decreased as the concentration of divalent ion was increased in a manner consistent with the negative surface potential theory of Lewis (1979 J. Physiol. (Lond.). 286: 417--445). With 20 mM XCl2 and 85.5 mM glucosamine.HCl in the external solution, the apparent permeability ratios for the alkaline earth cations (X++) are in the range 0.18--0.25. Alkali metal ions see the endplate channel as a water-filled, neutral pore without high-field-strength sites inside. Their permeability sequence is the same as their aqueous mobility sequence. Divalent ions, however, have a permeability sequence almost opposite from their mobility sequence and must experience some interaction with groups in the channel. In addition, the concentrations of monovalent and divalent ions are increased near the channel mouth by a weak negative surface potential.


Investigation of the composition and antioxidant activity of acetone and methanol extracts of Daphne sericea L. and Daphne gnidioides L.
The compositions of methanol and acetone extracts obtained from Daphne sericea L. and Daphne gnidioides L. were investigated. The antioxidant activities of each extract were determined by various test methods. Phenolic composition profile analysis by HPLC-DAD showed that D. gnidioides extracts contain more phenolic compounds than D. sericea extracts. Among the components, syringic acid was found to be the most abundant one in D. gnidioides extracts (42.8 and 38.4 mg per g dry extract of methanol and acetone, respectively). Total flavonoid, total phenolic and total carotenoid contents of methanolic D. gnidioides extracts were found to be 244.5 mg rutin/g dry weight of extract, 1219.3 mg GAE/g dry weight of extract and 11.9 mg/g dry weight of extract, respectively. DPPH·, ABTS·+ and O2·- assays were applied to plant extracts as radical scavenging activity tests. Methanolic extracts of D. sericea and D. gnidioides showed the highest radical scavenging activities according to DPPH· and ABTS·+ tests (61.6 and 147.2 in terms of IC50, respectively). Antioxidant activity tests for measuring lipid oxidation inhibiting capacity were carried in low density lipoprotein (LDL) and bovine brain extract (BBE). Methanolic extracts of D. gnidioides and D. sericea demonstrated remarkable lipid oxidation inhibiting capacity in LDL and BBE tests.


Blindness and visual impairment in western Bulgaria.
The Sofia Eye Survey was a population-based blindness and visual impairment survey of 6,275 randomly selected men and women from the Sofia district and Sofia city in western Bulgaria. A complete house-to-house census on all adults over age 40 years was performed, and visual acuity performed on 98%. We referred 171 subjects for a complete dilated ophthalmologic examination because the pinhole vision in the better eye was 6/18 or worse. Trained ophthalmologists refracted all referred subjects and then examined them with a slit lamp and ophthalmoscopy to determine the cause of visual loss. The rate of blindness (defined as vision in the better eye worse than 3/60) in this population was 0.49% and the rate of visual impairment (6/18 to 3/60 in the better eye) was 0.83%. The leading cause of both blindness and visual impairment was cataract. These data from the first community-based survey of blindness in Eastern Europe suggest that the rates of blindness and visual impairment are similar to those reported from other developed countries.


The nature and timing of coagulation dysfunction in a cohort of trauma patients in the Australian pre-hospital setting.
Acute Traumatic Coagulopathy (ATC) is a complex pathological process that is associated with patient mortality and increased blood transfusion requirements. It is evident on hospital arrival, but there is a paucity of information about the nature of ATC and the characteristics of patients that develop ATC in the pre-hospital setting. The objective of this study was to describe the nature and timing of coagulation dysfunction in a cohort of injured patients and to report on patient and pre-hospital factors associated with the development of ATC in the field. This was a prospective observational study of a convenience sample of trauma patients. Patients had blood taken during the pre-hospital phase of care and evaluated for derangements in Conventional Coagulation Assays (CCA) and Rotational Thromboelastometry (ROTEM). Associations between coagulation derangement and pre-hospital factors and patient outcomes were evaluated. A total of 216 patients who had either a complete CCA or ROTEM were included in the analysis. One hundred and eighty (83 %) of patients were male, with a median injury severity score of 17 [interquartile range (IQR) 10-27] and median age of 34 years [IQR = 25.0-52.0]. Hypofibrinogenemia was the predominant abnormality seen, (CCA Hypofibrinogenemia: 51/193, 26 %; ROTEM hypofibrinogenemia: 65/204, 32 %). Increased CCA derangement, the presence of ROTEM coagulopathy, worsening INR, worsening FibTEM and decreasing fibrinogen concentration, were all associated with both mortality and early massive transfusion. Clinically significant, multifaceted coagulopathy develops early in the clinical course, with hypofibrinogenemia being the predominant coagulopathy. In keeping with the ED literature, pre-hospital coagulation dysfunction was associated with mortality and early massive transfusion. Further work is required to identify strategies to identify and guide the pre-hospital management of the coagulation dysfunction seen in trauma.


Pathway-based biomarker search by high-throughput proteomics profiling of secretomes.
An efficient means for the identification of prognostic and predictive biomarkers is essential in today's cancer management. A new approach toward biomarker discovery has therefore been proposed, where pathways instead of individual proteins would be monitored and targeted. Recently, the 'secretome', a biological fluid that may be enriched with secreted and/or shed proteins from adjacent disease-relevant cancer cells, has been targeted for biomarker discovery. We describe a novel method for secretome analysis using "stacking gels", label-free relative quantitation, and pathway analysis. The protocol presented here increases the throughput of secretome analysis by approximately 1 order of magnitude compared to earlier methodologies. In the first application, six cancer cell lines from three different tissues were studied. The global secretome data sets obtained were analyzed using pathway analysis software to attempt integrating the experimental findings into a cellular signaling context. This suggested that several secretome proteins might be interconnected with intracellular canonical pathways. This, in turn, may eventually allow the use of secretomes for discovery of pathway-based biomarkers. When this strategy was applied to two breast cancer cell lines, it appeared that the IGF signaling and the plasminogen activating system may be differentially regulated in invasive breast cancer, but this remains speculative until it is verified in a clinical setting. In summary, the methodology proposed optimizes cell culture with sample fractionation and LC-MS to obtain the highest yield from cultured cell secretomes, with a focus on rational biomarker discovery through putative linkage with cancer relevant pathways.


Measurement of electrical current perception threshold of sensory nerves for pruritus in atopic dermatitis patients and normal individuals with various degrees of mild damage to the stratum corneum.
Patients with atopic dermatitis (AD) are well known to be sensitive to irritation from the environment due to the impaired function of the stratum corneum (SC). Electrical current perception threshold (CPT) evaluation quantifies the sensory threshold to transcutaneous electrical stimulation of the sensory nerves. To study the CPT in a noninvasive fashion using Neurometer CPT/C, together with measurements of various functions of the SC. We measured the CPT on the flexor forearm and cheek of AD patients and normal individuals. Subsequently, we evaluated the CPT and skin sensitivity to a 30% aqueous solution of lactic acid after the infliction of various mildly disruptive measures on the SC on the flexor forearm of healthy individuals by the following three methods: (1) removal of the superficial sebum with acetone/ether, (2) scarification with a needle and (3) tape stripping of the SC. Finally, we examined the effect of topical applications of emollients such as petrolatum or a moisturizing cream to the scratched skin. AD patients showed a lower barrier function and lower CPT than normal individuals. In subsequent studies conducted in normal individuals, the CPT was found to be inversely correlated with transepidermal water loss (TEWL) levels after tape stripping. However, most of all, the partial superficial scarification with a needle decreased the CPT and increased the lactic acid stinging response. Prolonged removal of lipids from the SC with acetone/ether for 30 min that increased the TEWL levels for only 1 day decreased the high-frequency conductance value for 2 days and the CPT only on the 2nd day after treatment. Topical applications of emollients were effective to prevent the increased sensitivity caused by scratching. AD patients showed functional abnormalities of the SC and tended to have more sensitive skin on the cheek and flexor forearm than healthy controls. Even focal SC damage caused by superficial cracking may lead to further disruption of the already damaged SC in AD patients, by eliciting scratching and facilitating the permeation of various environmental allergens and also the induction of hypersensitive skin.


Bone marrow abnormalities in HIV infected children, report of three cases and review of the literature.
Bone marrow abnormalities in HIV infected adults include hypocellularity, myelodysplasia and poor marrow recovery. Data in children is limited. We report a series of three HIV infected with varied bone marrow abnormalities. First child was a 7-year-old boy with pulmonary tuberculosis, anemia, thrombocytopenia and bone marrow examination showed hypoplastic marrow. He succumbed to his disease within seven days of hospitalization. Second child was a three and a half year old girl who had severe anemia and her bone marrow examination showed dyserythropoiesis. Third child was a 7-year-old boy who had splenic abscesses and pancytopenia and bone marrow examination showed myelofibrosis with increased plasma cells. He also succumbed due to a fatal pulmonary bleed. Thus, advanced HIV disease in children can lead to bone marrow suppression in form of hypoplasia or myelofibrosis which can be fatal.


The Role of the Pulmonary Embolism Response Team: How to Build One, Who to Include, Scenarios, Organization, and Algorithms.
Pulmonary embolism response teams (PERTs) are multidisciplinary response teams aimed at delivering a range of diagnostic and therapeutic modalities to patients with pulmonary embolism. These teams have gained traction on a national scale. However, despite sharing a common goal, individual PERT programs are quite individualized-varying in their methods of operation, team structures, and practice patterns. The tendency of such response teams is to become intensely structured, algorithmic, and inflexible. However, in their current form, PERT programs are quite the opposite. They are being creatively customized to meet the needs of the individual institution based on available resources, skills, personnel, and institutional goals. After a review of the essential core elements needed to create and operate a PERT team in any form, this article will discuss the more flexible feature development of the nascent PERT team. These include team planning, member composition, operational structure, benchmarking, market analysis, and rudimentary financial operations.


Hyperpolarized xenon in NMR and MRI.
Hyperpolarized gases have found a steadily increasing range of applications in nuclear magnetic resonance (NMR) and NMR imaging (MRI). They can be regarded as a new class of MR contrast agent or as a way of greatly enhancing the temporal resolution of the measurement of processes relevant to areas as diverse as materials science and biomedicine. We concentrate on the properties and applications of hyperpolarized xenon. This review discusses the physics of producing hyperpolarization, the NMR-relevant properties of 129Xe, specific MRI methods for hyperpolarized gases, applications of xenon to biology and medicine, polarization transfer to other nuclear species and low-field imaging.


Treatment of co-occurring posttraumatic stress disorder and substance use: Does order of onset influence outcomes?
Posttraumatic stress disorder (PTSD) and substance use disorders (SUDs) represent major public health concerns, particularly among veterans. They are associated with significant distress and impairment, and are highly comorbid. Little is known, however, about what role the temporal order of diagnostic onset may play in severity of presenting symptomatology and treatment outcomes. The aim of this study, therefore, was to examine treatment outcomes by order of onset. Participants were 46 U.S. military veterans (91.3% male) enrolled in a larger randomized controlled trial examining the efficacy of an integrated, exposure-based treatment (Concurrent Treatment of PTSD and Substance Use Disorders Using Prolonged Exposure; COPE). Participants were grouped into 2 categories: (a) primary PTSD (i.e., PTSD developed before the onset of SUD) or (b) primary SUD (i.e., SUD developed before the onset of PTSD). No significant associations between order of onset and baseline symptomatology were observed. The findings revealed that participants with primary PTSD were significantly more likely than participants with primary SUD to report higher levels of PTSD symptoms at the end of treatment. However, there was no effect of order of onset on SUD outcomes. The findings suggest that individuals with earlier PTSD onset are a particularly high-risk group in terms of their trauma-related symptoms. Implications for treatment of comorbid PTSD/SUD are discussed. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


The phenotype of human Th17 cells and their precursors, the cytokines that mediate their differentiation and the role of Th17 cells in inflammation.
T helper 17 (T(h)17) cells represent a new subset of CD4+ effector T cells which have been described in both mice and humans. However, some differences seem to exist between murine and human T(h)17 cells with regard to their features, origin and role in immunopathology. Murine T(h)17 cells share their developmental origin with Foxp3+ Treg cells, indeed naive T-cell precursors can be differentiated to regulatory T (Treg) cells by transforming growth factor-beta (TGF-beta) alone, whereas the contemporaneous presence of TGF-beta and IL-6 gives origin to T(h)17 cells. Human T(h)17 cells which consistently express the CC chemokine receptor 6 and the equivalent of the murine NK1.1, CD161, appear to exclusively originate in response to IL-1beta and IL-23 from a small subset of CD161+CD4+ T-cell precursors detectable in the thymus and in umbilical cord blood. These cells constitutively express the T(h)17-driving transcription factor retinoic acid-related orphan receptor (ROR)gamma t and the IL-23R and can also give origin to T(h)1 cells or T(h)2 cells under the appropriate polarizing conditions. By contrast, human CD161-naive T cells only give rise to T(h)1 and T(h)2 cells, but not T(h)17 cells. TGF-beta may not exert a direct critical role in human T(h)17 cell differentiation, but indirectly favours their development by inhibiting the development of T(h)1 cells, which are much more susceptible than T(h)17 cells to its suppressive activity on cell proliferation. Moreover, while murine T(h)17 are pathogenic in some murine models of autoimmunity where T(h)1 cells seem to play a protective role, both T(h)17 and T(h)1 certainly contribute to the pathogenesis of human autoimmune and other chronic inflammatory disorders.


Apical membrane antigen 1 mediates apicomplexan parasite attachment but is dispensable for host cell invasion.
Apicomplexan parasites invade host cells by forming a ring-like junction with the cell surface and actively sliding through the junction inside an intracellular vacuole. Apical membrane antigen 1 is conserved in apicomplexans and a long-standing malaria vaccine candidate. It is considered to have multiple important roles during host cell penetration, primarily in structuring the junction by interacting with the rhoptry neck 2 protein and transducing the force generated by the parasite motor during internalization. Here, we generate Plasmodium sporozoites and merozoites and Toxoplasma tachyzoites lacking apical membrane antigen 1, and find that the latter two are impaired in host cell attachment but the three display normal host cell penetration through the junction. Therefore, apical membrane antigen 1, rather than an essential invasin, is a dispensable adhesin of apicomplexan zoites. These genetic data have implications on the use of apical membrane antigen 1 or the apical membrane antigen 1-rhoptry neck 2 interaction as targets of intervention strategies against malaria or other diseases caused by apicomplexans.


Ocular Posterior Segment Involvement in Patients with Antiphospholipid Syndrome and Systemic Lupus Erythematosus.
Purpose: To describe posterior segment findings of antiphospholipid syndrome (APS) and compare them with systemic lupus erythematosus (SLE).Methods: A total of 11 patients with primary APS, 29 secondary APS patients, and 29 SLE patients without APS were included. All patients were referred from rheumatology clinic for detailed ophthalmologic examination. When patients had suspicious lesions, fundus fluorescein angiography was performed (n = 56).Results: The most common retinal examination finding was peripheral venous tortuosity (17.5%) in APS, which was not observed in SLE group. Common FFA findings were pigment epithelial window defects (10%) and vascular filling delays (7.5%) in APS, which were observed in 27.5% and 3.5% of patients with SLE consecutively.Conclusion: Venous tortuosity was significantly more in patients with APS. There was no significant difference for other ocular findings between the groups. Ocular complication rate was lower compared to earlier reports, probably due to better management of disease activity with current treatment protocols.


Ethanol-regulated gene expression of neuroendocrine specific protein in mice: brain region and genotype specificity.
Neuroendocrine specific protein or reticulon 1 (NSP/RTN1) was identified as a putative ethanol-regulated gene using mRNA differential display in mice genetically selected for severe ethanol withdrawal (withdrawal seizure-prone, WSP). One transcript of RTN1 (3.0 kb) showed a statistically significant increase (13%) in relative abundance in whole brain of ethanol-treated WSP mice but not in mice selected for resistance to ethanol withdrawal convulsions (WSR). We hypothesized that ethanol-induced regulation of gene expression of mRTN1 is specific to mice predisposed to exhibit severe ethanol withdrawal and that the gene might be regulated differentially in specific brain regions. WSP and WSR selected lines and DBA/2J and C57BL/6J inbred strains of mice were exposed to ethanol vapor or air for 72 h. mRNA steady-state expression of RTN1 was assessed in hippocampus, cortex, and cerebellum. Results indicated that the pattern of ethanol-induced changes in gene expression was dependent upon transcript size, brain region, and genotype. Modest increases in the relative abundance of both transcripts of RTN1 were observed in the hippocampus and cortex of all ethanol-treated mice. Results from cerebellum showed a moderate decrease in expression of RTN1 (3.0 kb transcript) in WSP and DBA/2J mice, but not in the mice resistant to ethanol withdrawal (C57BL/6J and WSR). These results suggest a genotype-specific effect of chronic ethanol exposure on steady-state mRNA levels of RTN1 in the cerebellum. Overall, the results indicate a complex pattern of ethanol-induced regulation of the putative mouse homologue of RTN1 and suggest that specific brain regional changes may be involved in the expression of physical dependence.


Suppression of myasthenia gravis by antigen-specific mucosal tolerance and modulation of cytokines and costimulatory factors.
We have shown that mucosal administration of recombinant fragments corresponding to the human acetylcholine receptor (AChR) alpha subunit suppresses chronic ongoing experimental autoimmune myasthenia gravis (EAMG) in rats. Treated animals exhibit a Th1 to Th2/Th3 shift in their cytokine profile and downregulation of costimulatory factors. However, application of a xenogeneic recombinant fragment may have limitations when considered as a possible approach for the treatment of MG in humans. We therefore tested the potential of a syngeneic fragment and of long synthetic peptides to suppress EAMG. We found that a syngeneic fragment corresponding to the extracellular region of the rat AChR alpha subunit was as effective as the formerly described human xenogeneic fragment in suppressing ongoing EAMG. This is encouraging in view of the potential use of mucosally administered recombinant AChR fragments for the treatment of MG in humans. However, in severely affected individuals, this antigen-specific approach may need to be supported by direct modulation of cytokines and costimulatory factors known to be involved in the pathogenesis of EAMG. To test the potential of this approach, myasthenic rats were injected by antibodies either to the proinflammatory cytokine IL-18 or to the costimulatory factor CD40L. These treatments act via different mechanisms, but both lead to the alleviation of clinical symptoms even when given at the chronic phase of EAMG. We suggest that antagonists to key cytokines and/or costimulatory factors be used to augment antigen-specific treatments of myasthenia such as mucosal administration of AChR recombinant fragments.


A synthetic oligonucleotide model for evaluating the oxidation and crosslinking propensities of natural furan-modified DNA.
We have previously developed a crosslinking methodology for oligonucleotides based on the incorporation of furan moieties, which can be selectively oxidised to reactive intermediates that will quickly react with the opposite bases in DNA, forming toxic interstrand crosslinks (ICLs). Furan moieties also occur in natural DNA, as a result of oxidative stress. Moreover, the furan-containing degradation product of this modified DNA-kinetin-has been found to display beneficial anti-ageing effects. To investigate the apparent discrepancy between the effects of the synthetic and the natural furan modifications in DNA, a quick and easy postsynthetic method providing access to the natural modification in short synthetic oligonucleotides was developed. On checking for potential crosslinking propensity, we found that the furan moiety does indeed undergo oxidation, in this way functioning as an important scavenger for oxidative stress. The reactive intermediate, however, was shown to degrade without producing toxic crosslinked products.


Apoptosis of ligamentous cells of the cranial cruciate ligament from stable stifle joints of dogs with partial cranial cruciate ligament rupture.
To describe the presence and amount of apoptotic ligamentous cells in different areas of partially ruptured canine cranial cruciate ligaments (prCCLs) and to compare these findings with apoptosis of ligamentous cells in totally ruptured cranial cruciate ligaments (trCCLs). 20 dogs with prCCLs and 14 dogs with trCCLs. Dogs with prCCLs or trCCLs were admitted to the veterinary hospital for stifle joint treatment. Biopsy specimens of the intact area of prCCLs (group A) and the ruptured area of prCCLs (group B) as well as specimens from trCCLs (group C) were harvested during arthroscopy. Caspase-3 and poly (ADP-ribose) polymerase (PARP) detection were used to detect apoptotic ligamentous cells by immunohistochemistry. No difference was found in the degree of synovitis or osteophytosis between prCCLs and trCCLs. No difference was found in degenerative changes in ligaments between groups A and B. A substantial amount of apoptotic cells could be found in > 90% of all stained slides. A correlation (r(s) = 0.71) was found between the number of caspase-3-and PARP-positive cells. No significant difference was found in the amount of apoptotic cells among the 3 groups. No significant correlation could be detected between the degree of synovitis and apoptotic cells or osteophyte production and apoptotic cells. The lack of difference between the 3 groups indicates that apoptosis could be a factor in the internal disease process leading to CCL rupture and is not primarily a consequence of the acute rupture of the ligament.


A randomized controlled trial to assess the efficacy and cost-effectiveness of urinary catheters with silver alloy coating in spinal cord injured patients: trial protocol.
Patients with non-acute spinal cord injury that carry indwelling urinary catheters have an increased risk of urinary tract infection (UTIs). Antiseptic Silver Alloy-Coated Silicone Urinary Catheters seems to be a promising intervention to reduce UTIs; however, actual evidence cannot be extrapolated to spinal cord injured patients. The aim of this trial is to make a comparison between the use of antiseptic silver alloy-coated silicone urinary catheters and the use of standard urinary catheters in spinal cord injured patients to prevent UTIs. The study will consist in an open, randomized, multicentre, and parallel clinical trial with blinded assessment. The study will include 742 spinal cord injured patients who require at least seven days of urethral catheterization as a method of bladder voiding. Participants will be online centrally randomized and allocated to one of the two study arms (silver alloy-coated or standard catheters). Catheters will be used for a maximum period of 30 days or removed earlier if the clinician considers it necessary. The main outcome will be the incidence of UTIs by the time of catheter removal or at day 30 after catheterization, the event that occurs first. Intention-to-treat analysis will be performed, as well as a primary analysis of all patients. The aim of this study is to assess whether silver alloy-coated silicone urinary catheters improve ITUs in spinal cord injured patients. ESCALE is intended to be the first study to evaluate the efficacy of the silver alloy-coated catheters in spinal cord injured patients. NCT01803919.


Workplace mental health: developing an integrated intervention approach.
Mental health problems are prevalent and costly in working populations. Workplace interventions to address common mental health problems have evolved relatively independently along three main threads or disciplinary traditions: medicine, public health, and psychology. In this Debate piece, we argue that these three threads need to be integrated to optimise the prevention of mental health problems in working populations. To realise the greatest population mental health benefits, workplace mental health intervention needs to comprehensively 1) protect mental health by reducing work-related risk factors for mental health problems; 2) promote mental health by developing the positive aspects of work as well as worker strengths and positive capacities; and 3) address mental health problems among working people regardless of cause. We outline the evidence supporting such an integrated intervention approach and consider the research agenda and policy developments needed to move towards this goal, and propose the notion of integrated workplace mental health literacy. An integrated approach to workplace mental health combines the strengths of medicine, public health, and psychology, and has the potential to optimise both the prevention and management of mental health problems in the workplace.