Evaluation of the degradation and metabolic effects of the gut peptide xenin on insulin secretion, glycaemic control and satiety.
Recently, glucagon-like peptide 1 (GLP1) and glucose-dependent insulinotropic polypeptide (GIP) have received much attention regarding possible roles in aetiology and treatment of type 2 diabetes. However, peptides co-secreted from the same enteroendocrine cells are less well studied. The present investigation was designed to characterise the in vitro and in vivo effects of xenin, a peptide co-secreted with GIP from intestinal K-cells. We examined the enzymatic stability, insulin-releasing activity and associated cAMP production capability of xenin in vitro. In addition, the effects of xenin on satiety, glucose homoeostasis and insulin secretion were examined in vivo. Xenin was time dependently degraded (t(1/2)=162±6 min) in plasma in vitro. In clonal BRIN-BD11 cells, xenin stimulated insulin secretion at 5.6 mM (P<0.05) and 16.7 mM (P<0.05 to P<0.001) glucose levels compared to respective controls. Xenin also exerted an additive effect on GIP, GLP1 and neurotensin-mediated insulin secretion. In clonal β-cells, xenin did not stimulate cellular cAMP production, alter membrane potential or elevate intra-cellular Ca(2)(+). In normal mice, xenin exhibited a short-acting (P<0.01) satiety effect at high dosage (500 nmol/kg). In overnight fasted mice, acute injection of xenin enhanced glucose-lowering and elevated insulin secretion when injected concomitantly or 30 min before glucose. These effects were not observed when xenin was administered 60 min before the glucose challenge, reflecting the short half-life of the native peptide in vivo. Overall, these data demonstrate that xenin may have significant metabolic effects on glucose control, which merit further study.


Leptin receptor gene polymorphisms and sex modify the association between acetaminophen use and asthma among young adults: results from two observational studies.
Epidemiologic studies have demonstrated associations between acetaminophen use and asthma. This investigation sought to determine whether sex modifies the acetaminophen-asthma association and whether leptin (LEP) and leptin receptor (LEPR) gene polymorphisms modulate the sex-specific associations. Data from the Isle of Wight birth cohort (IOW; n = 1456, aged 18 years) and Kuwait University Allergy (KUA; n = 1154, aged 18-26 years) studies were analyzed. Acetaminophen use and current asthma were self-reported. Genotype information for eighteen polymorphisms in LEP and LEPR genes were available in the IOW study. Associations between acetaminophen use and asthma were stratified by sex and genotype. Poisson regression models with robust variance estimation were evaluated to estimate adjusted prevalence ratios (aPR) and 95% confidence intervals (CI). Acetaminophen use was dose-dependently associated with an increased prevalence of current asthma in the IOW and KUA studies. In both studies, sex-stratified analysis showed that acetaminophen use was associated with asthma among males, but not in females (Pinteraction < 0.05). Moreover, a sex- and genotype-stratified analysis of the IOW data indicated that acetaminophen was associated with asthma to a similar extent among males and females carrying two common alleles of LEPR polymorphisms. In contrast, among those carrying at least one copy of the minor allele of LEPR polymorphisms, the magnitude of association between acetaminophen use and asthma was pronounced among males (aPR = 6.83, 95% CI: 2.87-16.24), but not among females (aPR = 1.22, 95% CI: 0.61-2.45). The identified sex-related effect modification of the acetaminophen-asthma association varied across LEPR genotypes, indicating that the sex-specific association was confined to individuals with certain genetic susceptibility. If the acetaminophen-asthma association is causal, then our findings will aid susceptibility-based stratification of at-risk individuals and augment preventive public health efforts.


Circulating levels of allopregnanolone in humans: gender, age, and endocrine influences.
Allopregnanolone is a neuroactive steroid involved in modulating behavioral functions, stress, and neuroendocrine axes in rats. Changes in plasma allopregnanolone levels throughout the menstrual cycle have been reported in healthy women, but there exists no information on the possible gender or age-related changes or on the source(s) of circulating allopregnanolone. The aim of the present study was to assess serum allopregnanolone concentrations according to gender, menstrual cycle, age, and menopause in normal men and women; serum progesterone (P) and dehydroepiandrosterone (DHEA) levels were evaluated in the same specimens. In addition, the possible source of circulating allopregnanolone in fertile women was investigated by using stimulatory and inhibitory endocrine tests acting on the ovary and/or adrenal cortex. The present study included 189 fertile women, 112 postmenopausal women, and 46 men. Serum steroid levels were determined after extraction, using specific RIAs. Allopregnanolone levels in fertile women in the follicular phase were similar to those in age-matched men; no significant difference was found between fertile women in the follicular phase and postmenopausal women. The highest levels were found in fertile women during the luteal phase (P < 0.01). An age-related decrease was observed in men (P < 0.01), but not in women. P and DHEA levels were significantly higher in women than in men and were higher in fertile women than in postmenopausal women (P < 0.01). Both P and DHEA showed an age-related decrease in men and women (P < 0.01). Serum allopregnanolone and P, but not DHEA, significantly increased in response to a GnRH test, whereas corticotropin-releasing factor and ACTH tests elicited a significant increase in allopregnanolone, P, and DHEA levels (P < 0.01). The suppression of adrenal steroidogenesis by dexamethasone markedly reduced both allopregnanolone and DHEA serum levels (P < 0.01). In conclusion, the present study demonstrated that although men show an age-related decrease, serum allopregnanolone levels in women do not change with age and correlate with P levels during the menstrual cycle and in response to endocrine tests. Ovary and adrenal cortex may be major sources of circulating allopregnanolone in fertile women.


A clinical longitudinal study of multiple sclerosis in Cantabria, Spain.
There are many studies that analyze the clinical course of multiple sclerosis (MS), but only a few of them observed patients from their first episode. We report a MS series of patients longitudinally evaluated from the onset of the disease and compare outcome between immunosuppressive treatment and non-treatment groups. Patients were included when their follow-up was at least three years and serial examination was available. Poser's criteria were used for diagnosis. We measured neurological impairment according to Kurtzke's expanded disability status scale (EDSS). Fifty-five MS patients were followed-up. Forty-six had clinically defined and nine clinically probable MS. Thirty-three patients had relapsing-remitting, 11 had secondary-progressive and 11 had primary-progressive MS. Mean age of onset of the disease was 31.1 years and mean duration of follow-up was seven years. Pyramidal weakness and sensory symptoms were the most common initial manifestations. Median time to reach EDSS-3 and EDSS-6 from onset was 4.5 and 7.5 years, respectively. From a total of 23 patients with follow-up of 10 or more years, 11 (48 %) had benign MS. The average time from onset of MS to secondary-progressive was 6.1 years. There were no significant differences between treatment and non-treatment patients. Longitudinal studies starting from early clinical course of MS are most useful in determining its semeiology, clinical pattern and outcome. Our results call for caution in initiating early immunosuppressive or immunomodulatory therapy.


Biochemical and biological markers: implications for epidemiologic studies.
Given that a major task for environmental epidemiology is to provide clear evidence of immediate and long-term health risks so that appropriate preventive measures can be taken, biochemical and biological markers of potentially hazardous environmental exposures are of great interest and possibly of great value. Such markers fall into two discrete classes: (1) those quantitatively related to the exposure itself, reflecting the magnitude of such exposures or the body burden of the pollutants, and (2) those markers that reflect the biological response to such exposures. In this paper we discuss the use of biochemical and biological markers in epidemiologic studies. Methods are presented for the use of markers to decrease misclassification errors in exposure studies. Relationships are derived that give minimum required values for laboratory sensitivity and specificity. Markers are also discussed in terms of some of the inherent problems in their use (e.g., ethical and legal considerations) and the likelihood of acceptance by participants in epidemiologic studies, researchers, regulators, and health professionals.


Antigen-experienced CD4 T cells display a reduced capacity for clonal expansion in vivo that is imposed by factors present in the immune host.
It is thought that protective immunity is mediated in part by Ag-experienced T cells that respond more quickly and vigorously than naive T cells. Using adoptive transfer of OVA-specific CD4 T cells from TCR transgenic mice as a model system, we show that Ag-experienced CD4 T cells accumulate in lymph nodes more rapidly than naive T cells after in vivo challenge with Ag. However, the magnitude of clonal expansion by Ag-experienced T cells was much less than that of naive T cells, particularly at early times after primary immunization. Ag-experienced CD4 T cells quickly reverted to the slower but more robust clonal expansion behavior of naive T cells after transfer into a naive environment. Conversely, the capacity for rapid clonal expansion was acquired by naive CD4 T cells after transfer into passively immunized recipients. These results indicate that rapid in vivo response by Ag-experienced T cells is facilitated by Ag-specific Abs, whereas the limited capacity for clonal expansion is imposed by some other factor in the immune environment, perhaps residual Ag.


Targeting the FcεRI Pathway as a Potential Strategy to Prevent Food-Induced Anaphylaxis.
Despite attempts to halt it, the prevalence of food allergy is increasing, and there is an unmet need for strategies to prevent morbidity and mortality from food-induced allergic reactions. There are no known medications that can prevent anaphylaxis, but several novel therapies show promise for the prevention of food-induced anaphylaxis through targeting of the high-affinity IgE receptor (FcϵRI) pathway. This pathway includes multiple candidate targets, including tyrosine kinases and the receptor itself. Small molecule inhibitors of essential kinases have rapid onset of action and transient efficacy, which may be beneficial for short-term use for immunotherapy buildup or desensitizations. Short courses of FDA-approved inhibitors of Bruton's tyrosine kinase can eliminate IgE-mediated basophil activation and reduce food skin test size in allergic adults, and prevent IgE-mediated anaphylaxis in humanized mice. In contrast, biologics may provide longer-lasting protection, albeit with slower onset. Omalizumab is an anti-IgE antibody that sequesters IgE, thereby reducing FcϵRI expression on mast cells and basophils. As a monotherapy, it can increase the clinical threshold dose of food allergen, and when used as an adjunct for food immunotherapy, it decreases severe reactions during buildup phase. Finally, lirentelimab, an anti-Siglec-8 antibody currently in clinical trials, can prevent IgE-mediated anaphylaxis in mice through mast cell inhibition. This review discusses these and other emerging therapies as potential strategies for preventing food-induced anaphylaxis. In contrast to other food allergy treatments which largely focus on individual allergens, blockade of the FcϵRI pathway has the advantage of preventing clinical reactivity from any food.


[Vaginal ulcers in Behçet disease].
Incidence of Behçet disease in USA is 0.12 in 100,000; but in Mexico there are just two isolated cases. This paper reports a case of a 30 year-old female, who, when 26, had vaginal ulcers. She received antimicrobial therapy during four years. In March 2007 she came to author's hospital with a cervical vaginitis diagnose study, and presenting thorax with venous collateral circulation secondary to superior and inferior cava vein thrombosis. Colposcopy and vaginal wall biopsy were practiced and found chronic ulcer with vascular damage; oral ulcers were found too, and Pathergy test was positive. Currently there is no ocular damage. Patient received azathioprine therapy without improvement, later she received anticoagulant therapy with corticoids and has good evolution.


Fully automatic identification of AC and PC landmarks on brain MRI using scene analysis.
We describe a method for identification of brain structures from MRI data sets. The bulk of the paper concerns an automatic system for finding the anterior and posterior commissures [(AC) and (PC)] in the midsagittal plane. These landmarks are key for the definition of the Talairach space, commonly used in stereotactic neurosurgery, in the definition of common coordinate systems for the pooling of functional positron emission tomography (PET) images and for neuroanatomy studies. The process works according to a step-by-step procedure: it first analyzes the skull limits. A grey-level histogram is then calculated and allows an automated selection of thresholds. Then, the interhemispheric plane is detected. Following an advanced scene analysis in the midsagittal plane for anatomical structures, the AC and the PC are identified. Experimentally, with a set of 200 patients, the process never failed. Its performances and limits are comparable to that of neuroanatomy experts. Those results are due to a high degree of robustness at each step of the program.


The Lived Experience of the Hospital Discharge "Plan": A Longitudinal Qualitative Study of Complex Patients.
Transitions in care are a high-risk time for patients. Complex patients account for the largest proportion of healthcare costs but experience lower quality and discontinuity of care. The experiences of complex patients can be used to identify gaps in hospital discharge practices and design interventions to improve outcomes. We used a case study approach with serial interviews and chart abstraction to explore the hospital discharge and transition experience over 6 weeks. Participants were recruited from a small hospital in Toronto that provides care to complex patients living with human immunodeficiency virus (HIV). Framework analysis was used to compare data across time-points and sources. Data were collected from 9 cases. Participants presented with complex medical and psychosocial challenges, including substance use (n = 9), mental health diagnoses (n = 8) and a mean of 5 medical comorbidities in addition to HIV. Data were analyzed and reported in 4 key themes: 1) social support; 2) discharge process and transition experience; 3) post-discharge follow-up; and 4) patient priorities. After hospital discharge, the complexity of participants' lives resulted in a change in priorities and subsequent divergence from the discharge plan. Despite the comprehensive discharge plans, with referrals designed to support their health and activities of daily living, participants experienced challenges with social support and referral uptake, resulting in a loss of stability achieved while in hospital. Further investigation and changes in practice are necessary to ensure that discharge plans for complex patients are realistic within the context of their lives outside of the hospital. Journal of Hospital Medicine 2017;12:5-10.


Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions.
Mitogen-activated protein (MAP) kinases comprise a family of ubiquitous proline-directed, protein-serine/threonine kinases, which participate in signal transduction pathways that control intracellular events including acute responses to hormones and major developmental changes in organisms. MAP kinases lie in protein kinase cascades. This review discusses the regulation and functions of mammalian MAP kinases. Nonenzymatic mechanisms that impact MAP kinase functions and findings from gene disruption studies are highlighted. Particular emphasis is on ERK1/2.


"It Is Not the Robot Who Learns, It Is Me." Treating Severe Dysgraphia Using Child-Robot Interaction.
Writing disorders are frequent and impairing. However, social robots may help to improve children's motivation and to propose enjoyable and tailored activities. Here, we have used the Co-writer scenario in which a child is asked to teach a robot how to write via demonstration on a tablet, combined with a series of games we developed to train specifically pressure, tilt, speed, and letter liaison controls. This setup was proposed to a 10-year-old boy with a complex neurodevelopmental disorder combining phonological disorder, attention deficit/hyperactivity disorder, dyslexia, and developmental coordination disorder with severe dysgraphia. Writing impairments were severe and limited his participation in classroom activities despite 2 years of specific support in school and professional speech and motor remediation. We implemented the setup during his occupational therapy for 20 consecutive weekly sessions. We found that his motivation was restored; avoidance behaviors disappeared both during sessions and at school; handwriting quality and posture improved dramatically. In conclusion, treating dysgraphia using child-robot interaction is feasible and improves writing. Larger clinical studies are required to confirm that children with dysgraphia could benefit from this setup.


Ten-Year Clinical Outcome After Circumferential Pulmonary Vein Isolation Utilizing the Hamburg Approach in Patients With Symptomatic Drug-Refractory Paroxysmal Atrial Fibrillation.
Circumferential pulmonary vein isolation (CPVI) is increasingly performed for the treatment of symptomatic drug-refractory paroxysmal atrial fibrillation. Long-term data for >10 years after CPVI are sparse. We investigated the long-term clinical outcome and progression of paroxysmal atrial fibrillation after double-lasso and 3-dimensional electroanatomical mapping-guided CPVI. From 2003 to 2004, 161 patients (mean age: 60±10 years) with symptomatic drug-refractory paroxysmal atrial fibrillation were prospectively enrolled and underwent electroanatomical mapping-guided CPVI. Right-sided and left-sided continuous circular lesions encircling the ipsilateral pulmonary veins were placed with irrigated radiofrequency energy. The procedural end point was the absence of pulmonary vein spikes 30 minutes after CPVI verified by 2 spiral catheters placed within the ipsilateral pulmonary veins (double-lasso technique). Major periprocedural complications occurred in 5 of 161 patients (3.1%). Follow-up was based on outpatient clinic visits, including Holter-ECGs and telephonic interviews. After a single procedure and median follow-up of 129.0 months (interquartile range, 124.7-133.1 months), stable sinus rhythm was present in 53 of 161 (32.9%) patients. Multiprocedural outcome after a mean of 1.73±0.9 procedures and a median follow-up of 123.4 months (interquartile range, 61.0-131.0 months) resulted in stable sinus rhythm in 101 of 161 patients (62.7%). Progression toward persistent atrial fibrillation was observed in 10 of 161 patients (6.2%). Ten-year single-procedural outcome of CPVI in patients with paroxysmal atrial fibrillation resulted in stable sinus rhythm in 32.9% and in 62.7% of patients after multiple procedures. The progression rate to persistent AF was remarkably low.


Infantile haemangiomas: a challenge in paediatric dermatology.
Infantile haemangiomas, common benign vascular tumours of childhood, are characterized by rapid growth during the first year of life and a slow regression that is usually completed at 7-10 years of age. These tumours are composed of endothelial cells with high mitotic rates and stromal components such as fibroblasts, mast cells and pericytes. Haemangiomas become a challenge when they are part of a syndrome, are located in certain areas of the body or when complications develop. The above-mentioned factors also influence the treatment modality used. However, although there remain many uncertainties regarding management, the beta-adrenergic receptor blocker propranolol is a promising new candidate for first-line systemic therapy. It produces such a dramatic and rapid response that the appearance of an infantile haemangioma should impart expeditious consideration of the risks and benefits of its use.


Immune checkpoint inhibitor therapy-associated graft intolerance syndrome in a failed kidney transplant recipient.
Immune checkpoint inhibitors (ICPIs) are monoclonal antibodies against inhibitory receptors on T cells resulting in anticancer activity. In kidney transplant (KT) recipients, ICPI use has been associated with acute allograft rejection. In failed allografts, however, the effects of ICPIs are unknown. We present a case of a 66-year-old man with a history of diabetes, renal cell cancer, left native nephrectomy, and end-stage kidney disease. He received a deceased donor KT which failed after 6 years due to biopsy-proven recurrent diabetic nephrosclerosis. He was started on hemodialysis and his immunosuppression was gradually weaned off. A year later, he was diagnosed with renal cell cancer in his right native kidney requiring nephrectomy. He later developed metastasis and was started on combination ICPIs. He developed hematuria, allograft pain, and malaise consistent with graft intolerance syndrome 28 days after starting ICPIs. Urine culture and cystoscopy were normal. A computed tomography scan of his abdomen revealed an enlarged allograft with patchy enhancement. After a multidisciplinary discussion, he underwent transplant nephrectomy. Histopathology showed chronic active T cell-mediated rejection. As ICPI use becomes prevalent, practitioners need to be aware of its potential complications among KT recipients both with functioning and failed allografts.


Effect of reducing inspired oxygen concentration on oxygenation parameters during general anaesthesia in horses in lateral or dorsal recumbency.
To compare the effects of two concentrations of oxygen delivered to the anaesthetic breathing circuit on oxygenation in mechanically ventilated horses anaesthetised with isoflurane and positioned in dorsal or lateral recumbency. Selected respiratory parameters and blood lactate were measured and oxygenation indices calculated, before and during general anaesthesia, in 24 laterally or dorsally recumbent horses. Horses were randomly assigned to receive 100% or 60% oxygen during anaesthesia. All horses were anaesthetised using the same protocol and intermittent positive pressure ventilation (IPPV) was commenced immediately following anaesthetic induction and endotracheal intubation. Arterial blood gas analysis was performed and oxygenation indices calculated before premedication, immediately after induction, at 10 and 45 min after the commencement of mechanical ventilation, and in recovery. During anaesthesia, the arterial partial pressure of oxygen was adequate in all horses, regardless of position of recumbency or the concentration of oxygen provided. At 10 and 45 min after commencing IPPV, the arterial partial pressure of oxygen was lower in horses in dorsal recumbency compared with those in lateral recumbency, irrespective of the concentration of oxygen supplied. Based on oxygenation indices, pulmonary function during general anaesthesia in horses placed in dorsal recumbency was more compromised than in horses in lateral recumbency, irrespective of the concentration of oxygen provided. During general anaesthesia, using oxygen at a concentration of 60% instead of 100% maintains adequate arterial oxygenation in horses in dorsal or lateral recumbency. However, it will not reduce pulmonary function abnormalities induced by anaesthesia and recumbency.


Application of mixture distributions for identifying thresholds of frequent and high inpatient mental health service use in longitudinal data.
There is a need for greater understanding about frequent and high use of inpatient mental health services, and those with ongoing increased needs. Most studies employ a threshold of frequent use (e.g. numbers of admissions) and high use (e.g. lengthy stays) without justification. To identify model-driven thresholds for frequent/high inpatient mental health service use and contrast characteristics of patients identified using various models and thresholds. Retrospective population-based study using 12 years of longitudinal data for 5631 patients admitted with a mental health diagnosis. Two-component negative binomial and poisson mixture (truncated/untruncated) models identified thresholds for frequent/high use in a 12-month period. The two-component negative binomial mixture model resulted in the best model fit. Using negative binomial-derived thresholds, 5.3% of patients had a period of frequent use (admitted six or more times), 15.8% of high use (hospitalised for 45 or more days) and 3.5% of heavy use (both frequent and high use). The prevalence of specific mental health disorders (e.g. mood disorder and schizophrenia) among frequent and high use cohorts varied across thresholds. This model-driven approach can be applied to identify thresholds in other cohorts. Threshold choice may depend on the magnitude and focus of potential interventions.


Hepatic cirrhosis, alcohol and haemorrhage.
Fifty consecutive patients with cirrhosis of the liver were investigated, including 29 with gastrointestinal haemorrhage. Over two-thirds of these patients gave a history of heavy alcohol intake. The necessity for fibre-endoscopy is stressed in that barium studies alone showed oesophageal varices in only 40 percent of patients and erosive lesions were often present. Gastric varices, demonstrated by gastrophotography in 25 percent of patients with portal hypertension, were not readily detectable by standard barium meal or endoscopy. Radio-isotope scanning of the liver and spleen was invaluable in supporting not only the diagnosis of cirrhosis, but also the presence of portal hypertension, since the spleen was palpable in only 50 percent of such patients.


Interaction of the ADRB2 gene polymorphism with childhood trauma in predicting adult symptoms of posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD), while highly prevalent (7.6% over a lifetime), develops only in a subset of trauma-exposed individuals. Genetic risk factors in interaction with trauma exposure have been implicated in PTSD vulnerability. To examine the association of 3755 candidate gene single-nucleotide polymorphisms with PTSD development in interaction with a history of childhood trauma. Genetic association study in an Ohio National Guard longitudinal cohort (n = 810) of predominantly male soldiers of European ancestry, with replication in an independent Grady Trauma Project (Atlanta, Georgia) cohort (n = 2083) of predominantly female African American civilians. Continuous measures of PTSD severity, with a modified (interview) PTSD checklist in the discovery cohort and the PTSD Symptom Scale in the replication cohort. Controlling for the level of lifetime adult trauma exposure, we identified the novel association of a single-nucleotide polymorphism within the promoter region of the ADRB2 (Online Mendelian Inheritance in Man 109690) gene with PTSD symptoms in interaction with childhood trauma (rs2400707, P = 1.02 × 10-5, significant after correction for multiple comparisons). The rs2400707 A allele was associated with relative resilience to childhood adversity. An rs2400707 × childhood trauma interaction predicting adult PTSD symptoms was replicated in the independent predominantly female African American cohort. Altered adrenergic and noradrenergic function has been long believed to have a key etiologic role in PTSD development; however, direct evidence of this link has been missing. The rs2400707 polymorphism has been linked to function of the adrenergic system, but, to our knowledge, this is the first study to date linking the ADRB2 gene to PTSD or any psychiatric disorders. These findings have important implications for PTSD etiology, chronic pain, and stress-related comorbidity, as well as for both primary prevention and treatment strategies.


New Uses of AbobotulinumtoxinA in Aesthetics.
BotulinumtoxinA (BoNT-A) is now widely established for the main approved indication of reducing glabellar lines, and is also widely used off-label to improve the appearance of wrinkles and lines in other parts of the face. The number of aesthetic procedures continues to increase as the patient population becomes more diverse, in particular with increasing numbers of people of color and men. Further developments in treatment may continue to expand the audience for BoNT-A by making procedures more comfortable and by delivering a more natural, less static appearance. These may be achieved through use of combinations of BoNT-A with other aesthetic procedures, tailoring the dose of toxin to the patient's muscle mass or by using novel injection and application techniques. Beyond amelioration of facial lines, encouraging results have been seen with the use of BoNT-A to improve the appearance of hypertrophic and keloid scars and even to prevent them. Studies have been conducted with scars in various parts of the body and further research is ongoing. Dermatological and other medical uses for BoNT-A are also active areas of research. Injections of BoNT-A have been shown to reduce signs and symptoms of acne, rosacea, and psoriasis, to reduce neuromuscular pain, and to bring about significant improvements in a number of rare diseases that are caused or exacerbated by hyperhidrosis. This paper reviews these new uses for BoNT-A, looking at the rationale for their use and discussing the results of published case studies and clinical trials. These areas have shown great promise to date, but more and larger clinical studies will be required before these treatments become a clinical reality. To this end details are also provided of clinical trials currently listed in the main clinical trials database to highlight research areas of particular interest.


Lived Experience of Women with Severe Dysmenorrhea.
The purpose of this study was to understand the meaning and nature of the lived experience of women with severe dysmenorrhea. Hermeneutic phenomenology by van Mannen and feminist philosophy informed this study involving 20 women who were 10~40 years old. Data was collected by using focus group interview 2 times from 10 women and in-depth interview from other 10 women from September to December, 2010. The essential themes were message from body, deconstruction of negative stereotyped body, and authorship of my body. Participants described their own painful experiences. They recognized that psychological stress impacted on severe dysmenorrhea, so they made self caring time. They had positive attitude to menstruation, said that dysmenorrhea was not illness, but normal life process. They had internal strength, wisdom, and sistership. This study revealed meaning of pain experience in sociocultural context. This finding have implications for health care provider's empathic and holistic practice.


Presurgical columellar elongation and primary retrograde nasal reconstruction in one-stage bilateral cleft lip and nose repair.
We present a new combined approach to primary bilateral cleft lip, nose, and alveolus repair using presurgical nasoalveolar molding combined with a one-stage lip, nose, and alveolus repair. Presurgical alveolar molding is used to bring the protruding premaxilla back into proper alignment with the lateral segments in the maxillary arch. Presurgical nasal molding produces tissue expansion of the short columella and nasal lining. A coordinated surgical approach involves a one-stage repair of the lip, nose, and alveolus. The nasal repair uses a retrograde approach in which the prolabial flap and columella are reflected over the nasal dorsum by continuing the dissection behind the prolabium up the membranous septum and over the septal angle. Tissues are dissected out from between the tip cartilages, and the domes are sutured together in the midline. This method joins a new class of bilateral cleft repairs that place the primary emphasis on correction of the deformity of the nasal tip cartilages.


Integrated assessment of west coast of South Korea by use of benthic bacterial community structure as determined by eDNA, concentrations of contaminants, and in vitro bioassays.
During the past few decades, contamination of sediments by persistent toxic substances (PTSs) has been observed in estuarine and coastal areas on the west coast of South Korea. The contaminants are suspected to cause toxicities in aquatic biota, but little is known about their ecological effects, particularly on benthic microbial communities. In this study, an eDNA-based assessment was applied along with classic assessments of exposure, such as chemistry and in vitro bioassays, to evaluate condition of benthic bacterial communities subjected to PTSs. Two strategies were adopted for the study. One was to conduct a comprehensive assessment in space (by comparing seawater and freshwater sites at five coastal regions) and in time (by following change over a 5-y period). Although we found that bacterial composition varied among and within years, some phyla, such as Proteobacteria (28.7%), Actinobacteria (13.1%), Firmicutes (12.7%), and Chloroflexi (12.5%) were consistently dominated across the study regions. Certain bacterial groups, such as Firmicutes and Verrucomicrobia have been linked to contamination at some sites in the study area and at specific points in time. Bacterial communities were not significantly correlated with salinity or AhR- and ER-mediated potencies, whereas concentrations of PAHs, APs, and certain metals (Cd and Hg) exhibited significant associations to the structure of bacterial communities at the phylum level. In fact, the relative abundance of microbes in the phylum Planctomycetes was significantly and negatively correlated with concentrations of PAHs and metals. Thus, the relative abundance of Planctomycetes could be used as an indicator of sedimentary contamination by PAHs and/or metals. Based on our correlation analyses, Cd and ER-mediated potencies were associated more with bacterial abundances at the class taxonomic level than were other PTSs and metals. Overall, the eDNA-based assessment was useful by augmenting more traditional measures of exposure and responses in a sediment triad approach and has potential as a more rapid screening tool.


Complexes between protein export chaperone SecB and SecA. Evidence for separate sites on SecA providing binding energy and regulatory interactions.
During localization to the periplasmic space or to the outer membrane of Escherichia coli some proteins are dependent on binding to the cytosolic chaperone SecB, which in turn is targeted to the membrane by specific interaction with SecA, a peripheral component of the translocase. Five variant forms of SecB, previously demonstrated to be defective in mediating export in vivo (Gannon, P. M., and Kumamoto, C. A. (1993) J. Biol. Chem. 268, 1590-1595; Kimsey, H. K., Dagarag, M. D., and Kumamoto, C. A. (1995) J. Biol. Chem. 270, 22831-22835) were investigated with respect to their ability to bind SecA both in solution and at the membrane translocase. We present evidence that at least two regions of SecA are involved in the formation of active complexes with SecB. The variant forms of SecB were all capable of interacting with SecA in solution to form complexes with stability similar to that of complexes between SecA and wild-type SecB. However, the variant forms were defective in interaction with a separate region of SecA, which was shown to trigger a change that was correlated to activation of the complex. The region of SecA involved in activation of the complexes was defined as the extreme carboxyl-terminal 21 aminoacyl residues.


Nanostructure of aluminum oxide inclusion and formation of hydrogen bubbles in molten aluminum.
Hydrogen in molten aluminum is one of the major factors that lead to pore formation in the solidification process of aluminum castings. Previous research showed that aluminum oxide inclusion had a close correlation with the hydrogen content in molten aluminum. Though some researchers thought there must have been a relationship between surface morphology of the inclusion and hydrogen concentration in molten aluminum, very few documents have reported on the surface property of aluminum oxide inclusion. In the present work, AFM (Atomic Force Microscope) was first used to investigate surface morphology of aluminum oxide inclusion in molten aluminum. It was found that there were a large number of nanoscale micropores on the surface of aluminum oxide inclusion. The interior of the micropores was approximated as a tapered shape. These micropores were thought to be helpful to heterogeneous nucleation for hydrogen atoms aggregation because they provided necessary concentration fluctuation and energy undulation for the growth of hydrogen bubbles. Based on the nanostructure observed on the surface of aluminum oxide inclusion, a theoretical model was developed to describe the hydrogen pore formation in aluminum casting under the condition of heterogeneous nucleation.


Use of complementary and alternative therapy by women in the first 2 years after diagnosis and treatment of invasive breast cancer.
Our aim was to document patterns of consultation with alternative practitioners, use of complementary and alternative medicine (CAM) commonly used to alleviate menopausal symptoms, and lifestyle changes made by women in the first 2 years after their diagnosis with invasive breast cancer. A total of 1,588 women in the MBF Foundation Health and Wellbeing After Breast Cancer Study completed both the enrollment questionnaire and the first follow-up questionnaire (FQ1), on average, 41 and 92 weeks after diagnosis, respectively. The Menopause Quality of Life Questionnaire and the Psychological General Wellbeing Index were included in the FQ1. Logistic regression analysis was used to examine characteristics associated with consulting a CAM practitioner or using CAMs. At the time of completion of the FQ1, 16.4% of women had consulted a CAM practitioner and 10.6% reported using at least one CAM commonly used to alleviate menopausal symptoms. Women who consulted a CAM practitioner and women who were using at least one CAM at the time of completion of the FQ1 were, on average, younger and more educated and had a higher Menopause Quality of Life Questionnaire score than did women who did not. Of the women, 33.6% reported having made a dietary change, and the most common lifestyle change was an increase in physical activity, which was reported by 29.0% of women. The frequency of use of CAMs in common usage for menopausal symptoms by women with breast cancer was lower than anticipated, increased with time from diagnosis, and is associated with more severe menopausal symptoms.


Quetiapine monotherapy as treatment for anxiety symptoms in patients with bipolar depression: a pooled analysis of results from 2 double-blind, randomized, placebo-controlled studies.
To evaluate the efficacy and tolerability of quetiapine monotherapy for anxiety symptoms in patients with bipolar disorder experiencing depression in the BipOLar DEpRession (BOLDER I and II) studies. A post hoc analysis of anxiety symptoms in 1,051 acutely depressed patients with bipolar I or II disorder (DSM-IV) from 2 double-blind, randomized, placebo-controlled 8-week studies of quetiapine (300 or 600 mg once daily) was conducted. Anxiety symptoms were assessed using Hamilton Anxiety Rating Scale (HARS) total and psychic (items 1-6, 14) and somatic (items 7-13) anxiety subscale scores (mixed-model repeated measure and last-observation-carried-forward analysis of change from baseline at each assessment). The BOLDER I study was conducted between September 2002 and October 2003, and the BOLDER II study was conducted between June 2004 and August 2005. Mean baseline HARS total scores were similar across the treatment groups (300 mg/d: 18.9, 600 mg/d and placebo: both 18.6). There was a significantly greater improvement from baseline in mean HARS total scores at the first evaluation (week 1) in both quetiapine groups compared with placebo (300 mg/d: -4.6, P < .001 and 600 mg/d: -4.1, P = .003 vs placebo: -2.8). These improvements were sustained through week 8 with both quetiapine doses (300 mg/d: -10.1, P < .001 and 600 mg/d: -10.5, P < .001 vs placebo: -6.9). At week 8, there was also significant improvement from baseline in HARS psychic and somatic anxiety subscale scores compared with placebo (P < .001). The baseline severity of anxiety did not impact the improvement in depressive symptoms. Common adverse events included dry mouth, sedation, somnolence, and dizziness. In this pooled analysis, quetiapine monotherapy was more effective than placebo and generally well tolerated for the treatment of both depressive and anxiety symptoms in patients with bipolar disorder. clinicaltrials.gov Identifiers: NCT00060489 (BOLDER I) and NCT00083954 (BOLDER II).


Novel marine actinobacteria from emerald Andaman & Nicobar Islands: a prospective source for industrial and pharmaceutical byproducts.
Andaman and Nicobar Islands situated in the eastern part of Bay of Bengal are one of the distinguished biodiversity hotspot. Even though number of studies carried out on the marine flora and fauna, the studies on actinobacteria from Andaman and Nicobar Islands are meager. The aim of the present study was to screen the actinobacteria for their characterization and identify the potential sources for industrial and pharmaceutical byproducts. A total of 26 actinobacterial strains were isolated from the marine sediments collected from various sites of Port Blair Bay where no collection has been characterized previously. Isolates were categorized under the genera: Saccharopolyspora, Streptomyces, Nocardiopsis, Streptoverticillium, Microtetraspora, Actinopolyspora, Actinokineospora and Dactylosporangium. Majority of the isolates were found to produce industrially important enzymes such as amylase, protease, gelatinase, lipase, DNase, cellulase, urease and phosphatase, and also exhibited substantial antibacterial activity against human pathogens. 77% of isolates exhibited significant hemolytic activity. Among 26 isolates, three strains (NIOT-VKKMA02, NIOT-VKKMA22 and NIOT-VKKMA26) were found to generate appreciable extent of surfactant, amylase, cellulase and protease enzyme. NIOT-VKKMA02 produced surfactant using kerosene as carbon source and emulsified upto E(24)-63.6%. Moreover, NIOT-VKKMA02, NIOT-VKKMA22 and NIOT-VKKMA26 synthesized 13.27 U/ml, 9.85 U/ml and 8.03 U/ml amylase; 7.75 U/ml, 5.01 U/ml and 2.08 U/ml of cellulase and 11.34 U/ml, 6.89 U/ml and 3.51 U/ml of protease enzyme, respectively. High diversity of marine actinobacteria was isolated and characterized in this work including undescribed species and species not previously reported from emerald Andaman and Nicobar Islands, including Streptomyces griseus, Streptomyces venezuelae and Saccharopolyspora salina. The enhanced salt, pH and temperature tolerance of the actinobacterial isolates along with their capacity to secrete commercially valuable primary and secondary metabolites emerges as an attractive feature of these organisms. These results are reported for the first time from these emerald Islands and expand the scope to functionally characterize novel marine actinobacteria and their metabolites for the potential novel molecules of commercial interest.


Acquired warfarin resistance and weight-reducing diet.
A frequent cause of acquired warfarin resistance is drug interaction; however, ingestion of large amounts of vitamin K in food may also be an etiologic factor. A 31-year-old woman on a weight-reducing diet showed evidence of resistance to warfarin sodium therapy. On a regimen of 35 mg of oral warfarin sodium daily, prothrombin time was 14 s (control, 12 s). Pharmacokinetic studies did not reveal any evidence of impaired adsorption or increased catabolism of the drug. The half-life of her oral warfarin was 26 hours (normal, 15 to 56 hours). Although end-organ resistance was not studied fully, a change of her vegetable-rich, weight-reducing diet (vitamin K content, 1,277 microgram) to a regular diet (vitamin K content, 360 microgram) resulted in substantial reduction in her warfarin resistance. We conclude that in patients on vegetable-rich, weight-reducing diets, a relative resistance to warfarin may be secondary to their increased dietary intake of vitamin K.


Broad-range glycosidase activity profiling.
Plants produce hundreds of glycosidases. Despite their importance in cell wall (re)modeling, protein and lipid modification, and metabolite conversion, very little is known of this large class of glycolytic enzymes, partly because of their post-translational regulation and their elusive substrates. Here, we applied activity-based glycosidase profiling using cell-permeable small molecular probes that react covalently with the active site nucleophile of retaining glycosidases in an activity-dependent manner. Using mass spectrometry we detected the active state of dozens of myrosinases, glucosidases, xylosidases, and galactosidases representing seven different retaining glycosidase families. The method is simple and applicable for different organs and different plant species, in living cells and in subproteomes. We display the active state of previously uncharacterized glycosidases, one of which was encoded by a previously declared pseudogene. Interestingly, glycosidase activity profiling also revealed the active state of a diverse range of putative xylosidases, galactosidases, glucanases, and heparanase in the cell wall of Nicotiana benthamiana. Our data illustrate that this powerful approach displays a new and important layer of functional proteomic information on the active state of glycosidases.


Healing of a Chronic Pressure Injury in a Patient Treated With Medical Cannabis for Pain and Sleep Improvement: A Case Report.
A small body of evidence suggests medical cannabis may facilitate wound healing, but the exact mechanism of this effect is unclear. This case report describes a patient with a pressure injury (PI) who received cannabis oil treatment for pain management and sleep improvement. A 37-year-old woman with multiminicore disease, scoliosis, short-chain acyl-CoA dehydrogenase deficiency, and epilepsy presented to the Neurology Centre of Toronto with chronic pain and sleep disturbance, including difficulty initiating and maintaining sleep. She also had a 5-year history of a PI between her right iliac crest and right rib cage that had progressively worsened. The patient received a medical cannabis oil protocol that used a combination of cannabidiol and tetrahydrocannabinol. Cannabis oil was effective in treating pain and sleep difficulties. Unexpectedly, during the first 2 weeks of treatment, the PI started to heal and was almost completely closed at the 2-month follow-up. Although it is unknown if the observed healing of this refractory PI was indirectly or directly related to the cannabidiol and tetrahydrocannabinol treatment, the potential relationships among pain, sleep disturbance, cannabis treatment, and healing should be explored.


Immunologic constant of rejection signature is prognostic in soft-tissue sarcoma and refines the CINSARC signature.
Soft-tissue sarcomas (STSs) are heterogeneous and aggressive tumors, with high metastatic risk. The immunologic constant of rejection (ICR) 20-gene signature is a signature of cytotoxic immune response. We hypothesized that ICR might improve the prognostic assessment of early-stage STS. We retrospectively applied ICR to 1455 non-metastatic STS and searched for correlations between ICR classes and clinicopathological and biological variables, including metastasis-free survival (MFS). Thirty-four per cent of tumors were classified as ICR1, 27% ICR2, 24% ICR3, and 15% ICR4. These classes were associated with patients' age, pathological type, and tumor depth, and an enrichment from ICR1 to ICR4 of quantitative/qualitative scores of immune response. ICR1 class was associated with a 59% increased risk of metastatic relapse when compared with ICR2-4 class. In multivariate analysis, ICR classification remained associated with MFS, as well as pathological type and Complexity Index in Sarcomas (CINSARC) classification, suggesting independent prognostic value. A prognostic clinicogenomic model, including the three variables, was built in a learning set (n=339) and validated in an independent set (n=339), showing greater prognostic precision than each variable alone or in doublet. Finally, connectivity mapping analysis identified drug classes potentially able to reverse the expression profile of poor-prognosis tumors, such as chemotherapy and targeted therapies. ICR signature is independently associated with postoperative MFS in early-stage STS, independently from other prognostic features, including CINSARC. We built a robust prognostic clinicogenomic model integrating ICR, CINSARC, and pathological type, and suggested differential vulnerability of each prognostic group to different systemic therapies.


Thermal-exfoliated synthesis of N-rich carbon-based nanosheets from layered bulk crystals of a metal-hexamine framework.
We develop a general strategy for the design of a series of 2D N-rich carbon-based nanomaterials through the thermal exfoliation of layered metal-hexamine framework microcrystals. By a facile pyrolysis, the sharply generated gases escape from the layered precursors, leading to the exfoliation of the layers and successful preparation of 2D carbon-based nanomaterials. This strategy can readily skip the complicated morphology engineering process of 2D metal-organic frameworks to produce 2D N-rich carbon-based nanomaterials on a large scale.


Feasibility of external beam radiation therapy to deep-seated targets with kilovoltage x-rays.
Radiation therapy to deep-seated targets is typically delivered with megavoltage x-ray beams generated by medical linear accelerators or 60 Co sources. Here, we used computer simulations to design and optimize a lower energy kilovoltage x-ray source generating acceptable dose distributions to a deep-seated target. The kilovoltage arc therapy (KVAT) x-ray source was designed to treat a 4-cm diameter target located at a 10-cm depth in a 40-cm diameter homogeneous cylindrical phantom. These parameters were chosen as an example of a clinical scenario for testing the performance of the kilovoltage source. A Monte Carlo (MC) model of the source was built in the EGSnrc/BEAMnrc code and source parameters, such as beam energy, tungsten anode thickness, beam filtration, number of collimator holes, collimator hole size and thickness, and source extent were varied. Dose to the phantom was calculated in the EGSnrc/DOSXYZnrc code for varying treatment parameters, such as the source-to-axis distance and the treatment arc angle. The quality of dose distributions was quantified by means of target-to-skin ratio and dose output expressed in D50 (50% isodose line) for a 30-min irradiation in the homogeneous phantom as well as a lung phantom. Additionally, a patient KVAT dose distribution to a left pararenal lesion (~1.6 cm in diameter) was calculated and compared to a 15 MV volumetric modulated arc therapy (VMAT) plan. In the design of the KVAT x-ray source, the beam energy, beam filtration, collimator hole size, source-to-isocenter distance, and treatment arc had the largest effect on the source output and the quality of dose distributions. For the 4-cm target at 10-cm depth, the optimized KVAT dose distribution generated a conformal plan with target-to-skin ratio of 5.1 and D50 in 30 min of 24.1 Gy in the homogeneous phantom. In the lung phantom, a target-to-skin ratio of 7.5 and D50 in 30 min of 25.3 Gy were achieved. High dose conformity of the 200 kV KVAT left pararenal plan was comparable to the 15 MV VMAT plan. The volume irradiated to at least 10% (<240 cGy) of the prescription dose was 2.2 × larger in the 200 kV KVAT plan than in the 15 MV VMAT plan, but considered clinically insignificant. This study demonstrated that conformal treatments of deep-seated targets were achievable with kilovoltage x-rays with dose distributions comparable to MV beams. However, due to the larger volumes irradiated to clinically tolerated low doses, KVAT x-ray source usage for deep-seated lesions will be further evaluated to determine optimal target size.


Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons.
Accurate modeling of human neuronal cell biology has been a long-standing challenge. However, methods to differentiate human induced pluripotent stem cells (iPSCs) to neurons have recently provided experimentally tractable cell models. Numerous methods that use small molecules to direct iPSCs into neuronal lineages have arisen in recent years. Unfortunately, these methods entail numerous challenges, including poor efficiency, variable cell type heterogeneity, and lengthy, expensive differentiation procedures. We recently developed a new method to generate stable transgenic lines of human iPSCs with doxycycline-inducible transcription factors at safe-harbor loci. Using a simple two-step protocol, these lines can be inducibly differentiated into either cortical (i3 Neurons) or lower motor neurons (i3 LMN) in a rapid, efficient, and scalable manner (Wang et al., 2017). In this manuscript, we describe a set of protocols to assist investigators in the culture and genetic engineering of iPSC lines to enable transcription factor-mediated differentiation of iPSCs into i3 Neurons or i3 LMNs, and we present neuronal culture conditions for various experimental applications. © 2018 by John Wiley & Sons, Inc.


Serologic Testing of US Blood Donations to Identify Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)-Reactive Antibodies: December 2019-January 2020.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease 2019 (COVID-19), was first identified in Wuhan, China, in December 2019, with subsequent worldwide spread. The first US cases were identified in January 2020. To determine if SARS-CoV-2-reactive antibodies were present in sera prior to the first identified case in the United States on 19 January 2020, residual archived samples from 7389 routine blood donations collected by the American Red Cross from 13 December 2019 to 17 January 2020 from donors resident in 9 states (California, Connecticut, Iowa, Massachusetts, Michigan, Oregon, Rhode Island, Washington, and Wisconsin) were tested at the Centers for Disease Control and Prevention for anti-SARS-CoV-2 antibodies. Specimens reactive by pan-immunoglobulin (pan-Ig) enzyme-linked immunosorbent assay (ELISA) against the full spike protein were tested by IgG and IgM ELISAs, microneutralization test, Ortho total Ig S1 ELISA, and receptor-binding domain/ACE2 blocking activity assay. Of the 7389 samples, 106 were reactive by pan-Ig. Of these 106 specimens, 90 were available for further testing. Eighty-four of 90 had neutralizing activity, 1 had S1 binding activity, and 1 had receptor-binding domain/ACE2 blocking activity >50%, suggesting the presence of anti-SARS-CoV-2-reactive antibodies. Donations with reactivity occurred in all 9 states. These findings suggest that SARS-CoV-2 may have been introduced into the United States prior to 19 January 2020.


Leprosy in Elderly and Children among New Cases - A 3-Year Retrospective Study.
Leprosy occurs in all age groups, with adults constituting the majority. However, leprosy in children always drew the attention of epidemiologists as a guide to transmission of leprosy. With increasing life expectancy and decreasing prevalence of leprosy, there is going to be a significant rise of leprosy among elderly in India. In elderly leprosy patients, clinical signs are often quiet, which makes it a hidden source of infection. The detection of leprosy in elderly is of epidemiological importance, hence it is critical that due attention be given to leprosy in elderly as a possible contributor to hidden leprosy in India. To analyze leprosy in elderly and in children from the records of new patients seen over the past 3-year period. Analysis of details of "leprosy in elderly" and "leprosy in children'" from the records of new leprosy patients seen at a dermatology OPD of a teaching hospital over a period of 3 years. Out of 157 patients, 34 (21.6%) were above 45 years, while 18 (11.4%) were below 15 years, the difference being statistically significant (P > 0.05). Bacteriological Index (BI) values in elderly were higher compared to children. In addition, elderly had a higher percentage of multibacillary (MB) leprosy compared to children both clinically (35% vs 22%) and histopathologically (38% vs 22%). Leprosy in elderly is an important entity, and there is a need to study it as a distinct group. It will provide information on hidden leprosy load and apprise us on sources of infection in the community.


Management of the posttraumatic arthritic knee.
Excellent long-term outcomes can be achieved with contemporary methods of ligament reconstruction and open reduction and internal fixation for injuries around the knee; nevertheless, posttraumatic arthritis frequently develops. Reconstruction options for symptomatic posttraumatic knee arthritis include osteotomy, arthrodesis, and arthroplasty. Surgical challenges include the presence of extensive (often broken) hardware, scarring, stiffness, bony defects, compromised soft tissues, and malalignment. Patient age and activity and the anatomic location and extent of damage to the articular surface must be taken into account when determining the surgical treatment plan. For younger patients, osteotomy, allograft transplantation, or arthrodesis of the knee is considered, whereas older, low-demand patients are usually treated with arthroplasty. Attention to specific technical details and careful surgical technique are necessary to achieve a successful result. Functional improvement is usually seen following arthroplasty and, sometimes, arthrodesis. However, complications are common, and outcomes following arthroplasty are generally inferior to those reported for other diagnoses.


Protein L: a robust enzyme-conjugated molecule for detection of humanized single chain antibodies.
The current study was aimed at introducing an efficient enzyme-conjugated molecule for use in the detection of humanized single chain antibodies. Because they are composite in their variable domain sequences and also devoid of constant domains, humanized single chain antibodies require a suitable secondary enzyme-conjugated antibody (more precisely enzyme-conjugated protein molecule) to be efficiently recognized and detected in ELISA, dot blot, and other detection tests, guaranteeing a more precise evaluation of their quantity, affinity, and other features. In the current study we examined the ability of three HRP-conjugated protein molecules including protein L, anti-human IgG antibody, and anti-mouse IgG antibody in the detection of three humanized single chain antibodies (anti-CD20, anti-EGFRvIII, and anti-EGFR) in ELISA and dot blot tests. The results signify the HRP-conjugated protein L as the most efficient molecule for detecting humanized single chain antibody.


Lidocaine increases intracellular sodium concentration through voltage-dependent sodium channels in an identified lymnaea neuron.
The local anesthetic lidocaine affects neuronal excitability in the central nervous system; however, the mechanisms of such action remain unclear. The intracellular sodium concentration ([Na]i) and sodium currents (INa) are related to membrane potential and excitability. Using an identifiable respiratory pacemaker neuron from Lymnaea stagnalis, the authors sought to determine whether lidocaine changes [Na]i and membrane potential and whether INa is related to these changes. Intracellular recording and sodium imaging were used simultaneously to measure membrane potentials and [Na]i, respectively. Measurements for [Na]i were made in normal, high-Na, and Na-free salines, with membrane hyperpolarization, and with tetrodotoxin pretreatment trials. Furthermore, changes of INa were measured by whole cell patch clamp configuration. Lidocaine increased [Na]i in a dose-dependent manner concurrent with a depolarization of the membrane potential. In the presence of high-Na saline, [Na]i increased and the membrane potential was depolarized; the addition of lidocaine further increased [Na]i, and the membrane potential was further depolarized. In Na-free saline or in the presence of tetrodotoxin, lidocaine did not change [Na]i. Similarly, hyperpolarization of the membrane by current injections also prevented the lidocaine-induced increase of [Na]i. In the patch clamp configuration, membrane depolarization by lidocaine led to an inward sodium influx. A persistent reduction in membrane potential, resulting from lidocaine, brings the cell within the window current of INa where sodium channel activation occurs. Lidocaine increases intracellular sodium concentration and promotes excitation through voltage-dependent sodium channels by altering membrane potential in the respiratory pacemaker neuron.


Engineering a mobile platform to promote sleep in the pediatric primary care setting.
Pediatricians lack tools to support families at home for the promotion of childhood sleep. We are using the Multiphase Optimization Strategy (MOST) framework to guide the development of a mobile health platform for childhood sleep promotion. The objective of this study is to demonstrate feasibility of a mobile health platform towards treating children with insufficient sleep. Children aged 10-12 years were enrolled (Study #1: N = 30; Study #2: N = 43). Participants wore a sleep tracker to measure sleep duration. Data were retrieved by a mobile health platform, programmed to send introductory messages during run-in (2 weeks) and goal achievement messages during intervention (7 weeks) periods. In study #1, participants were randomized to control, gain-framed incentive or loss-framed incentive arms. In study #2, participants were randomized to control, loss-framed incentive, normative feedback or loss-framed incentive plus normative feedback arms. In study #1, 1514 nights of data were captured (69%) and sleep duration during the intervention was higher by an average of 21 (95% CI: -8, 51) and 34 (95% CI: 7, 61) minutes per night for the gain-framed and loss-framed arms, respectively, compared to controls. In study #2, 2,689 nights of data were captured (81%), with no major differences in average sleep duration between the control and the loss-framed or normative feedback arms. We have developed and deployed a mobile health platform that can capture sleep data and remotely communicate with families. Promising candidate intervention components will be further investigated under the optimization phase of the MOST framework. Both studies included in this manuscript were registered at clinicaltrials.gov:-Study #1: NCT03263338-Study #2: NCT03426644.


Melatonin attenuates sevoflurane-induced hippocampal damage and cognitive deficits in neonatal mice by suppressing CypD in parvalbumin neurons.
Sevoflurane, a commonly administered inhaled anesthetic, is found to induce synaptic and mitochondrial damage in neonatal mice. Mitochondrial membrane potential (MMP) changes, mediated by Cyclophilin D (CypD), are implicated in mitochondrial function. Melatonin, known for its significant neuroprotective properties, was investigated in this study to elucidate its mechanisms in mitigating the cognitive impairment caused by sevoflurane. The mice were categorized into several groups, including the control, vehicle, sevoflurane, vehicle plus sevoflurane, and melatonin plus sevoflurane groups. From postnatal day 6 to day 8, the mice were administered inhaled sevoflurane or intraperitoneal melatonin. MMP and reactive oxygen species (ROS) were measured using appropriate detection kits. The protein expression levels of PSD95, Synapsin Ⅰ, and CypD in the hippocampus were analyzed through western blotting in acute and prolonged terms. Immunofluorescence staining was used to assess the co-localizations of PSD95 or CypD in parvalbumin (PV) neurons. Cognitive ability was evaluated through novel object recognition, social interaction experiment, and the Morris water maze. The findings revealed that repeated exposure to sevoflurane in neonatal mice resulted in cognitive and synaptic impairment. Furthermore, melatonin administration suppressed the ROS and CypD protein expression, enhanced the MMP in mitochondria and synaptic protein expression in PV neurons, and ameliorated cognitive deficits. Melatonin alleviated sevoflurane-induced cognitive deficits by suppressing CypD and promoting synaptic development in hippocampal PV neurons. These results provide valuable insights into a promising therapeutic approach for preventing neurotoxic injuries caused by general anesthetics.


CD38 protein deficiency induces autoimmune characteristics and its activation enhances IL-10 production by regulatory B cells.
CD38 is a transmembrane protein expressed in B lymphocytes, and is able to induce responses as proliferation, differentiation or apoptosis. Several reports propose that CD38 deficiency accelerates autoimmune processes in murine models of autoimmune diabetes, lymphoproliferation and rheumatoid arthritis. Other reports have shown elevated CD38 expression in B and T cells from patients with autoimmunity; however, the role of CD38 is still unclear in the development of autoimmunity. Recently, it has been characterized as CD1dhi CD5+ regulatory B cell subpopulation able to produce IL-10, and the loss of these cells exacerbates the autoimmunity in murine models. Here, we report that CD38-/- mice exhibited elevated titres of ANAS, anti-dsDNA autoantibodies from 12 months of age and were higher by 16 months of age and mice presented kidney damage. Interestingly, there is a reduction in the survival of CD38-/- mice compared to the WT. Furthermore, CD38 is highly expressed by CD1dhigh CD5+ regulatory B cells, and the agonistic anti-CD38 stimulus plus LPS was able to increase the percentage of this cell subset and its ability to induce IL-10 production. Together, these results suggest that CD38 could play a role in the control of autoimmune diseases through their expression on regulatory B cells.


Increased photoreversal of ultraviolet injury by flashing light.
1. Photoreversal of ultraviolet (UV) injury was studied in the ciliate protozoan Tetrahymena pyriformis (geleii) strain W, cultured in the absence of other living organisms. The division pattern of progeny of single animals was followed in hanging drop preparations. 2. A sublethal dose of 450 ergs/mm.(2) of monochromatic UV of wave length 2654 A produces a lag before the first division followed by a period of cessation of fission after the second division. This cessation sometimes lasts as long as 6 weeks, during which time the animals become smaller and rounder and more opaque. Organisms about to resume division increase in size and transparency; after a few divisions the animals regain their normal division rate. 3. The effect of UV ranging in intensity from 5 to 15 ergs/mm.(2)/sec. was found to obey the reciprocity law quite well for the UV effect on the division pattern of T. pyriformis. However, the same dose at lower and at higher intensities was less effective. 4. The effect of a dose of UV delivered at high intensity (19 ergs/mm.(2)/sec.) could be increased by flashing the light, indicating that the system became saturated in the continuous light. 5. A photoreversing dose of monochromatic blue light of wave length 4350 A was found to be more effective when delivered as continuous light at a low intensity, or as intermittent light at a high intensity, rather than as continuous light at the high intensity-indicating that a dark mechanism participates in photoreversal. 6. The time for the dark reaction was determined to be of the order of a few hundredths of a second in experiments in which different lengths of dark period were used while maintaining a constant light period of 0.0025 second. 7. For Colpidium colpoda the efficiency of a given dose of photoreversing light was increased by flashing the light. 8. The present experiments are interpreted in terms of data available in the literature.


Nanomaterials for brain metastasis.
Tumor metastasis is a significant contributor to the mortality of cancer patients. Specifically, current conventional treatments are unable to achieve complete remission of brain metastasis. This is due to the unique pathological environment of brain metastasis, which differs significantly from peripheral metastasis. Brain metastasis is characterized by high tumor mutation rates and a complex microenvironment with immunosuppression. Additionally, the presence of blood-brain barrier (BBB)/blood tumor barrier (BTB) restricts drug leakage into the brain. Therefore, it is crucial to take account of the specific characteristics of brain metastasis when developing new therapeutic strategies. Nanomaterials offer promising opportunities for targeted therapies in treating brain metastasis. They can be tailored and customized based on specific pathological features and incorporate various treatment approaches, which makes them advantageous in advancing therapeutic strategies for brain metastasis. This review provides an overview of current clinical treatment options for patients with brain metastasis. It also explores the roles and changes that different cells within the complex microenvironment play during tumor spread. Furthermore, it highlights the use of nanomaterials in current brain treatment approaches.


Parent and household influences on calcium intake among early adolescents.
Calcium intake during early adolescence falls short of requirements for maximum bone accretion. Parents and the home food environment potentially influence children's calcium intakes. This study aimed to quantify parental psychosocial factors (PSF) predicting calcium intakes of Asian, Hispanic, and non-Hispanic white (NHW) early adolescent children from a parental perspective. This was a cross-sectional study involving the administration of a validated calcium-specific food frequency questionnaire to a convenience sample of children aged 10-13 years and the primary individual responsible for food acquisition in the child's household. Based on Social Cognitive Theory, parental factors potentially associated with children's calcium intake were also assessed via parent questionnaires. The total study sample consisted of 633 parent-child pairs (Asian = 110, Hispanic = 239, NHW = 284). Questionnaires were completed at community-based centers/sites. Outcome measures were the association between parent-child calcium (mg), milk (cups/day), and soda (cans/day) intakes and the predictive value of significant parental PSF towards calcium intakes of their children. Sex-adjusted linear regression and multivariate analyses were performed. Calcium intakes of parent-child pairs were positively associated among all ethnic groups (r = 0.296; P < 0.001). Soda intakes were positively associated among Hispanic parent-child pairs only (r = 0.343; P < 0.001). Home availability of calcium-rich foods (CRF), parental rules and expectations for their child's intake of beverages, and parents' calcium intake/role modeling were positively associated with children's calcium intake and overwhelmed all other PSF in multivariate analyses. Significant cultural differences were observed. Parental role modeling was a significant factor among Hispanic dyads only. Multivariate models explained 19-21% of the variance in children's calcium intakes. Nutrition interventions to improve children's calcium intakes should focus on parents and provide guidance on improving home availability of CRF and increasing rules and expectations for the consumption of CRF. Among Hispanic families, interventions promoting parental modeling of desired dietary behaviors may be most successful.


Rapid prototyping of heterostructured organic microelectronics using wax printing, filtration, and transfer.
Conducting polymers are the natural choice for soft electronics. However, the main challenge is to pattern conducting polymers using a simple and rapid method to manufacture advanced devices. Filtration of conducting particle dispersions using a patterned membrane is a promising method. Here, we show the rapid prototyping of various micropatterned organic electronic heterostructures of PEDOT:PSS by inducing the formation of microscopic hydrogels, which are then filtered through membranes containing printed hydrophobic wax micropatterns. The hydrogels are retained on the un-patterned, hydrophilic regions, forming micropatterns, achieving a resolution reaching 100 μm. We further solve the problem of forming stacked devices by transferring the acidified PEDOT:PSS micropattern using the adhesive tape transfer method to form vertical heterostructures with other micropatterned electronic colloids such as CNTs, which are patterned using a similar technique. We demonstrate a number of different heterostructure devices including micro supercapacitors and organic electrochemical transistors and also demonstrate the use of acidified PEDOT:PSS microstructures in cell cultures to enable bioelectronics.


[Radiologic aspects of the congenital arteriovenous malformations, Klippel-Trenaunay type, and Servelle-Martorell type (author's transl)].
In our study 13 patients with a Klippel-Trenaunay type and 7 patients with a Servelle-Martorell type of arteriovenous malformations were analyzed. The results demonstrate that these 2 entities can be differentiated by routine radiography. The Klippel-Trenaunay type of angiodysplasia does not need arteriography, whereas arteriography as well as phlebography are necessary in the Servelle-Martorell type of angiodysplasia, in order to show ectatic regions of the involved vessels. The differentiation between the above mentioned arteriovenous malformations and the F. P. Weber type of angiodysplasia is easy. In the F. P. Weber type of angiodysplasia a lengthening of the involved extremity, arteriovenous shunts, as well as alterations of the bone are characteristic.


Scorpion β-toxin interference with NaV channel voltage sensor gives rise to excitatory and depressant modes.
Scorpion β toxins, peptides of ∼70 residues, specifically target voltage-gated sodium (Na(V)) channels to cause use-dependent subthreshold channel openings via a voltage-sensor trapping mechanism. This excitatory action is often overlaid by a not yet understood depressant mode in which Na(V) channel activity is inhibited. Here, we analyzed these two modes of gating modification by β-toxin Tz1 from Tityus zulianus on heterologously expressed Na(V)1.4 and Na(V)1.5 channels using the whole cell patch-clamp method. Tz1 facilitated the opening of Na(V)1.4 in a use-dependent manner and inhibited channel opening with a reversed use dependence. In contrast, the opening of Na(V)1.5 was exclusively inhibited without noticeable use dependence. Using chimeras of Na(V)1.4 and Na(V)1.5 channels, we demonstrated that gating modification by Tz1 depends on the specific structure of the voltage sensor in domain 2. Although residue G658 in Na(V)1.4 promotes the use-dependent transitions between Tz1 modification phenotypes, the equivalent residue in Na(V)1.5, N803, abolishes them. Gating charge neutralizations in the Na(V)1.4 domain 2 voltage sensor identified arginine residues at positions 663 and 669 as crucial for the outward and inward movement of this sensor, respectively. Our data support a model in which Tz1 can stabilize two conformations of the domain 2 voltage sensor: a preactivated outward position leading to Na(V) channels that open at subthreshold potentials, and a deactivated inward position preventing channels from opening. The results are best explained by a two-state voltage-sensor trapping model in that bound scorpion β toxin slows the activation as well as the deactivation kinetics of the voltage sensor in domain 2.


The Relationships among Tryptophan, Kynurenine, Indoleamine 2,3-Dioxygenase, Depression, and Neuropsychological Performance.
It has been suggested that the metabolic enzyme indoleamine 2,3-dioxygenase (IDO) is a biological mediator of inflammation related to the psychopathology of depression, with a Kynurenine (KYN) increase in the Tryptophan (TRP) metabolic pathway, resulting in reduced Serotonin. In this study, we examined KYN, TRP, and the ratio of KYN to TRP concentrations × 103 (KT Ratio) in serum and cerebrospinal fluid (CSF) in (a) a group of depressed patients and (b) a control group of patients referred to a neurologic outpatient clinic for whom no specific diagnosis could be established. The KT Ratio is considered an index that represents IDO. The participants were examined with the Beck Depression Inventory II (BDI-II), the Montgomery Aasberg Depression Rating Scale (MADRS), and a neuropsychological test battery. We found no significant differences between the two study groups with respect to TRP, KYN, or KT Ratio in serum or CSF. Differences in neuropsychological performance between the two patient groups could be seen in the following tests: Animal Fluency, Digit Symbol, the DKEFS Color-Interference Test (Naming Part), Trail Making Test A and B, and the Grooved Pegboard Non-dominant Hand. KYN in serum correlated highly with KYN in CSF. KYN in serum correlated significantly with both age and gender. When analyzing males and females separately, we found that women had a lower level of TRP in both serum (Mann-Whitney U-test: TRP in Serum; p = 0.001) and CSF (Mann-Whitney U-test: TRP in CSF; p = 0.003). Women had a lower level of KYN in serum (p = 0.029) than men did. Age was positively associated with KYN. KYN in CSF correlated only with age, however; there were no gender differences. No significant relationship was seen between BDI-II and MADRS on the one hand, and KYN and TRP on the other. KYN in CSF as the KT Ratio in both serum and CSF was associated with neuropsychological performance. Thus, we suggest that KYN and KT Ratio are related more strongly to neuropsychological performance than to affective symptoms in depression.


Patulin in apple juices: Incidence and likely intake in an Indian population.
Fifty samples of apple juices were investigated for patulin levels using HPLC with UV detection. It was found that 38 (76? samples contained no detectable patulin (LOD = 7.5 µg l(-1)), but some samples had high patulin levels (845 µg l(-1)) compared to other branded mixed juices (21-70 µg l(-1)) and apple juices procured from local vendors (191 µg l(-1)). The survey provides evidence of a need to educate the apple-juice industry in the use of good quality apples for juice production. In addition, the levels of patulin found showed a saturation of the provisional maximum tolerable daily intake (PMTDI) based on intake figures in terms of an average and 90th percentile of 27-60% and 107-190%, respectively, in different age groups. This monitoring study may help in formulating guidelines for patulin levels, which are not yet implemented by Prevention of Food Adulteration (PFA) Act of India.


A literature review of consent declines and consent withdrawals in randomized controlled trials conducted during the COVID-19 pandemic.
We evaluated the extent of consent declines and consent withdrawals during the COVID-19 pandemic as seen in published randomized controlled trials (RCTs) and compared it with non-COVID-19 RCTs published at the same time and two historical controls. PubMed/Medline only was searched using key-word "COVID-19" and "RCTs" separately, and filtered for COVID-19 RCTs and non-COVID-19 RCTs respectively, published during a nine-month period (1 Feb - 1 Nov 2020). Exclusions were study protocols, observational studies, interim analysis of RCT data and RCTs with missing data. Primary outcome measures were the proportion of consent declines and consent withdrawals as percentage of total participants screened and randomized respectively in COVID-19 RCTs. We compared consent declines and consent withdrawals of COVID-19 RCTs with non-COVID-19 RCTs and two earlier studies on the same topic that served as historical controls (non-pandemic setting). The search yielded a total of 111 COVID-19 RCTs and 49 non-COVID-19 RCTs. Of these, 39 (35.13%) COVID-19 RCTs and 11 (22.45%) non-COVID-19 RCTs were finally analysed. A total of 770/17759 (4.3%) consent declines and 100/7607 (1.31%) consent withdrawals were seen in 39 COVID-19 RCTs. A significant difference was observed in consent declines between COVID-19 vs non-COVID-19 RCTs [4.3% vs 11.9%, p < 0.0001] and between COVID-19 RCTs vs two historical controls [(4.3% vs 8.6%, p < 0.0001) and (4.3% vs 21.1%, p < 0.0001), respectively]. RCTs conducted during the COVID-19 pandemic appear to have significantly lower consent declines relative to non-COVID-19 RCTs during pandemic and RCTs conducted in non-pandemic settings.


[Volume of surgical resection by liver focal lesions].
Results of surgical treatment of 109 patients with focal lesions of the liver were analyzed. Volume resection by malignant and benign liver lesions was defined, depending on size, localization, number of foci and functional liver reserve. Postoperative lethality of patients with colorectal cancer metastases was 4.3%. Method of surgical treatment depended directly on the tumor stage. Overall 5-year survival by stage IVA hepatocellular cancer was 48%, 3.5-year survival was 62%. Overall and disease-free survival by colorectal cancer metastases was 35.1 and 22.4%, respectively.


Combination radioimmunotherapy and chemoimmunotherapy involving different or the same targets improves therapy of human pancreatic carcinoma xenograft models.
Chemoimmunotherapy with antibody-drug conjugates (ADC) is emerging as a promising therapy for solid tumors, whereas radioimmunotherapy (RAIT) of solid tumors has been relatively ineffective because of their resistance to radiation. We developed antibody-SN-38 conjugates that have significant antitumor activity in xenograft models at nontoxic doses. The goal of this study was to determine if an ADC could be combined with RAIT to enhance efficacy without a commensurate increase in host toxicity. Nude mice bearing human pancreatic cancer xenografts (Capan-1 and BxPC-3) were treated with a single dose of 90Y-labeled antimucin antibody (hPAM4; clivatuzumab tetraxetan) alone or in combination with an anti-Trop-2-SN-38 conjugate, typically administered twice weekly over 4 weeks. The combination, even at RAIT's maximum tolerated dose, controlled tumor progression and cured established xenografts significantly better than the individual treatments without appreciable toxicity. The ADC could be started 1 week after or up to 2 weeks before RAIT with similar enhanced responses, but delaying RAIT for 2 weeks after the ADC was less effective. A nonspecific ADC provided additional benefit over using free drug (irinotecan), but the response was enhanced with the specific ADC. When targeting Capan-1 with ample mucin, hPAM4 could be used as the RAIT and the ADC agent without losing effectiveness, but in BxPC-3 with less mucin, targeting of different antigens was preferred. These studies show the feasibility of combining ADC and RAIT for improved efficacy without increased toxicity.


Molecular cytogenetic characterization of two cases with constitutional distal 11q duplication/triplication.
Here, we report two cases with isolated distal 11q rearrangement and multiple congenital anomalies. The first patient is a two-and-a-half year old male referred to our genetics clinic due to dysmorphic features and developmental delay including speech delay. Using conventional and molecular cytogenetic techniques, we demonstrate that he carries a recombinant chromosome with duplication of the 11q23.3q24.2 region resulting from an intrachromosomal insertion in the father. The second patient was originally reported by Partida-Perez, et al. [Partida-Perez et al., 2006] as having a tandem duplication of the 11q23.3 region. We performed array comparative genomic hybridization (aCGH) on this patient in order to map the exact region of the duplication, and demonstrated that the patient actually had a triplication within 11q23.3. We compare the clinical features of our two patients with those previously reported to further delineate the phenotype of isolated distal 11q duplication. Our study also demonstrates the clinical usefulness of whole genome high resolution aCGH analysis as a powerful molecular cytogenetic tool capable of detecting genomic imbalances due to cytogenetically visible but uncertain rearrangements.


GMI, an immunomodulatory protein from Ganoderma microsporum, induces autophagy in non-small cell lung cancer cells.
Autophagy is a self-digestive process that degrades the cytoplasmic constituents. Immunomodulatory protein, one major bioactive component of Ganoderma, has antitumor activity. In this study, recombinant fungal immunomodulatory protein, GMI, was cloned from Ganoderma microsporum and purified. We demonstrated that GMI induces lung cancer cell death by activating autophagy, but does not induce apoptotic cell death. On western blot, GMI increased LC3 conversion and decreased p53 expression in a time- and concentration-dependent manner. Cytoplasmic calcium chelator BAPTA-AM was used to prove that GMI promotes autophagy via a calcium-mediated signaling pathway. 3-methyladenine (3-MA), an autophagy inhibitor, enhanced the cytotoxicity of GMI on cell viability assay. Using VZV-G pseudotyped lentivirus-shRNA system for autophagy-related genes silencing, the capabilities of GMI to reduce cell viability and colony formation were abolished in autophagy-defective cells. Furthermore, GMI did not stimulate apoptosis after blocking of autophagy by 3-MA or shRNA knockdown system. In xenograft studies, oral administration of GMI inhibited the tumor growth and induced autophagy significantly in nude mice that had received a subcutaneous injection of A549 cells. This is the first study to reveal the novel function of GMI in activating autophagy. GMI may be a potential chemopreventive agent against non-small cell lung cancer.


Enhancement of the virulence of Aeromonas caviae diarrhoeal strains by serial passages in mice.
Thirteen clinical Aeromonas caviae isolates from the faeces of 13 children with mild to severe diarrhoea were tested for enhancement of mouse lethality, adhesion ability, siderophore and cholera toxin cross-reactive (CTC) factor production, by four consecutive passages through mice by intraperitoneal injection of A. caviae suspensions. The passaged A. caviae strains were re-isolated from monomicrobic cardiac blood samples and inocula were prepared for the next passage. All A. caviae isolates possessed the ability to adhere to the mucosal epithelial surface of the rabbit small intestine. Serial passage in mice showed that the virulence of some isolates for mice was increased in terms of percentage mortality and a lowering of the LD50. For some of the isolates, but not all, serial passage appeared to increase siderophore production and adhesion to rabbit small intestinal cells. For the A. caviae isolates tested, increased values of the CTC factor were observed after passage. A clear correlation was observed between the lowering of LD50 and the enhancement of CTC factor production after passage in mice. These results indicate that the A. caviae isolates possessed virulence factors.


Rapid and efficient radiosynthesis of [123I]I-PK11195, a single photon emission computed tomography tracer for peripheral benzodiazepine receptors.
[(123)I]I-PK11195 is a high-affinity single photon emission computed tomography radiotracer for peripheral benzodiazepine receptors that has previously been used to measure activated microglia and to assess neuroinflammation in the living human brain. This study investigates the radiosynthesis of [(123)I]I-PK11195 in order to develop a rapid and efficient method that obtains [(123)I]I-PK11195 with a high specific activity for in vivo animal and human imaging studies. The synthesis of [(123)I]I-PK11195 was evaluated using a solid-state interhalogen exchange method and an electrophilic iododestannylation method, where bromine and trimethylstannyl derivatives were used as precursors, respectively. In the electrophilic iododestannylation method, the oxidants peracetic acid and chloramine-T were both investigated. Electrophilic iododestannylation produced [(123)I]I-PK11195 with a higher isolated radiochemical yield and a higher specific activity than achievable using the halogen exchange method investigated. Using chloramine-T as oxidant provided a rapid and efficient method of choice for the synthesis of [(123)I]I-PK11195. [(123)I]I-PK11195 has been successfully synthesized via a rapid and efficient electrophilic iododestannylation method, producing [(123)I]I-PK11195 with a higher isolated radiochemical yield and a higher specific activity than previously achieved.


Environmental enrichment promotes neural plasticity and cognitive ability in fish.
Different kinds of experience during early life can play a significant role in the development of an animal's behavioural phenotype. In natural contexts, this influences behaviours from anti-predator responses to navigation abilities. By contrast, for animals reared in captive environments, the homogeneous nature of their experience tends to reduce behavioural flexibility. Studies with cage-reared rodents indicate that captivity often compromises neural development and neural plasticity. Such neural and behavioural deficits can be problematic if captive-bred animals are being reared with the intention of releasing them as part of a conservation strategy. Over the last decade, there has been growing interest in the use of environmental enrichment to promote behavioural flexibility in animals that are bred for release. Here, we describe the positive effects of environmental enrichment on neural plasticity and cognition in juvenile Atlantic salmon (Salmo salar). Exposing fish to enriched conditions upregulated the forebrain expression of NeuroD1 mRNA and improved learning ability assessed in a spatial task. The addition of enrichment to the captive environment thus promotes neural and behavioural changes that are likely to promote behavioural flexibility and improve post-release survival.


An immunohistochemical and molecular analysis of papillary proliferation of the endometrium.
Papillary proliferations of the endometrium (PPEs) are uncommon lesions that are often associated with endometrial polyps. PPEs occasionally precede or co-exist with atypical endometrial hyperplasia or adenocarcinoma, but their pathogenesis and relationship to endometrial neoplasia is uncertain. In the present study 11 PPEs, including eight benign papillary proliferations (BPPs) and three complex papillary hyperplasias (CPHs) were examined immunohistochemically for expression of PAX2, BAF250a, p16, β-catenin and DNA mismatch repair (MMR) proteins. Molecular analysis was also performed on the CPHs using targeted next generation sequencing (NGS). All PPEs demonstrated at least one immunohistochemical abnormality with altered expression of p16 and PAX2 in nine and seven cases, respectively, and β-catenin in one case. However, none of the cases showed loss of BAF250a or MMR protein staining. All CPHs showed KRAS mutations with additional mutations in AKT1 and FBXW7 in one case each, and both PIK3CA and CTNNB1 in the remaining case. Therefore, PPEs demonstrate immunophenotypical and molecular overlap with endometrial endometrioid neoplasia, although loss of BAF250a and MMR protein function do not appear to contribute significantly to these lesions. KRAS mutations may be important drivers in CPHs but this finding needs to be confirmed in larger studies.


Red Clover (Trifolium pratense L.) Sprout Prevents Metabolic Syndrome.
We examined the prevention effect of red clover (Trifolium pratense L.) sprout on metabolic syndrome using a high-carbohydrate and high-fat diet (Western diet; WD)-induced male C57BL/6J obese model mouse. Red clover sprout-lyophilized powder (RC) contained 3.5 mg/g dry-weight of formononetin as a major phenolic compound, as analyzed by high performance liquid chromatography. Supplementation of 0.3% (w/w) RC in a WD (WD+RC) showed an anti-obesity effect and ameliorated lipid metabolism in the obese model mice. Additionally, fasting plasma glucose levels were significantly reduced in the WD+RC group. Administration of 0.1 mg/kg formononetin reduced the postprandial blood glucose level, as assessed using the oral maltose tolerance test. However, no significant formononetin intake effect was observed on the plasma insulin level. These results suggest that the formononetin contained in red clover sprout inhibits α-glucosidase and thereby contributes to reducing the postprandial blood glucose response in mice.


CCR5-edited gene therapies for HIV cure: Closing the door to viral entry.
Human immunodeficiency virus (HIV) was first reported and characterized more than three decades ago. Once thought of as a death sentence, HIV infection has become a chronically manageable disease. However, it is estimated that a staggering 0.8% of the world's population is infected with HIV, with more than 1 million deaths reported in 2015 alone. Despite the development of effective anti-retroviral drugs, a permanent cure has only been documented in one patient to date. In 2007, an HIV-positive patient received a bone marrow transplant to treat his leukemia from an individual who was homozygous for a mutation in the CCR5 gene. This mutation, known as CCR5Δ32, prevents HIV replication by inhibiting the early stage of viral entry into cells, resulting in resistance to infection from the majority of HIV isolates. More than 10 years after his last dose of anti-retroviral therapy, the transplant recipient remains free of replication-competent virus. Multiple groups are now attempting to replicate this success through the use of other CCR5-negative donor cell sources. Additionally, developments in the use of lentiviral vectors and targeted nucleases have opened the doors of precision medicine and enabled new treatment methodologies to combat HIV infection through targeted ablation or down-regulation of CCR5 expression. Here, we review historical cases of CCR5-edited cell-based therapies, current clinical trials and future benefits and challenges associated with this technology.


[Transfusion therapy of traumatic shock using plasma substitutes oxyamal and volecam].
The work is devoted to the investigation of efficiency of antishock transfusion therapy by new soviet drugs on the basis of hydroxyethyl starch--Oxyamal and Volecam. The investigation was performed on 30 patients with trauma shock and hypovolemic syndrome. The clinico-laboratory data obtained have shown that transfusion of Oxyamal and Volecam allow to rapidly normalize disturbed parameters of the central hemodynamics and continuously maintain them at the due level.


The chronic kidney disease - Mineral bone disorder (CKD-MBD): Advances in pathophysiology.
The causes of excess cardiovascular mortality associated with chronic kidney disease (CKD) have been attributed in part to the CKD-mineral bone disorder syndrome (CKD-MBD), wherein, novel cardiovascular risk factors have been identified. New advances in the causes of the CKD-MBD are discussed in this review. They demonstrate that repair and disease processes in the kidneys release factors to the circulation that cause the systemic complications of CKD. The discovery of WNT inhibitors, especially Dickkopf 1 (Dkk1), produced during renal repair as participating in the pathogenesis of the vascular and skeletal components of the CKD-MBD implied that additional pathogenic factors are critical. This lead to the discovery that activin A is a second renal repair factor circulating in increased levels during CKD. Activin A derives from peritubular myofibroblasts of diseased kidneys, wherein it stimulates fibrosis, and decreases tubular klotho expression. Activin A binds to the type 2 activin A receptor, ActRIIA, which is variably affected by CKD in the vasculature. In diabetic/atherosclerotic aortas, specifically in vascular smooth muscle cells (VSMC), ActRIIA signaling is inhibited and contributes to CKD induced VSMC dedifferentiation, osteogenic transition and neointimal atherosclerotic calcification. In nondiabetic/nonatherosclerotic aortas, CKD increases VSMC ActRIIA signaling, and vascular fibroblast signaling causing the latter to undergo osteogenic transition and stimulate vascular calcification. In both vascular situations, a ligand trap for ActRIIA prevented vascular calcification. In the skeleton, activin A is responsible for CKD stimulation of osteoclastogenesis and bone remodeling increasing bone turnover. These studies demonstrate that circulating renal repair and injury factors are causal of the CKD-MBD and CKD associated cardiovascular disease.