Regulation of relaxin 3 gene expression via cAMP-PKA in a neuroblastoma cell line.
Relaxin 3 is expressed in neurons of the brain stem that inneravate wide areas of the forebrain. Relaxin 3 mRNA levels in these neurons are increased in response to restraint stress, and by central administration of corticotropin-releasing factor (CRF). In the present study, we observed that relaxin 3 was expressed in a mouse neuroblastoma cell line, Neuro2a, and investigated the intracellular signaling that activated relaxin 3 gene transcription in vitro. By means of a clone stably transfected with a relaxin 3 promoter-EGFP gene, we observed that dibutyryl cyclic AMP and forskolin increased the relaxin 3 promoter activity. These increases were inhibited by pretreatment with PKA inhibitors, H89 and KT5720. Moreover, the promoter activity was enhanced by CRF treatment after expression of CRF-R1 receptor on the cells. Taken together, these results indicate that relaxin 3 transcription is activated via the cAMP-PKA pathway in the downstream of CRF-R1.


Roles for the cytoplasmic tails of the fusion and hemagglutinin-neuraminidase proteins in budding of the paramyxovirus simian virus 5.
The efficient release of many enveloped viruses from cells involves the coalescence of viral components at sites of budding on the plasma membrane of infected cells. This coalescence is believed to require interactions between the cytoplasmic tails of surface glycoproteins and the matrix (M) protein. For the paramyxovirus simian virus 5 (SV5), the cytoplasmic tail of the hemagglutinin-neuraminidase (HN) protein has been shown previously to be important for normal virus budding. To investigate a role for the cytoplasmic tail of the fusion (F) protein in virus assembly and budding, we generated a series of F cytoplasmic tail-truncated recombinant viruses. Analysis of these viruses in tissue culture indicated that the cytoplasmic tail of the F protein was dispensable for normal virus replication and budding. To investigate further the requirements for assembly and budding of SV5, we generated two double-mutant recombinant viruses that lack 8 amino acids of the predicted 17-amino-acid HN protein cytoplasmic tail in combination with truncation of either 10 or 18 amino acids from the predicted 20-amino-acid F protein cytoplasmic tail. Both of the double mutant recombinant viruses displayed a replication defect in tissue culture and a budding defect, the extent of which was dependent on the length of the remaining F cytoplasmic tail. Taken together, this work and our earlier data on virus-like particle formation (A. P. Schmitt, G. P. Leser, D. L. Waning, and R. A. Lamb, J. Virol. 76:3953-3964, 2002) suggest a redundant role for the cytoplasmic tails of the HN and F proteins in virus assembly and budding.


Estimated GFR and the Effect of Intensive Blood Pressure Lowering After Acute Intracerebral Hemorrhage.
The kidney-brain interaction has been a topic of growing interest. Past studies of the effect of kidney function on intracerebral hemorrhage (ICH) outcomes have yielded inconsistent findings. Although the second, main phase of the Intensive Blood Pressure Reduction in Acute Cerebral Hemorrhage Trial (INTERACT2) suggests the effectiveness of early intensive blood pressure (BP) lowering in improving functional recovery after ICH, the balance of potential benefits and harms of this treatment in those with decreased kidney function remains uncertain. Secondary analysis of INTERACT2, which randomly assigned patients with ICH with elevated systolic BP (SBP) to intensive (target SBP<140mmHg) or contemporaneous guideline-based (target SBP<180mmHg) BP management. 2,823 patients from 144 clinical hospitals in 21 countries. Admission estimated glomerular filtration rates (eGFRs) of patients were categorized into 3 groups based on the CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration) creatinine equation: normal or high, mildly decreased, and moderately to severely decreased (>90, 60-90, and <60mL/min/1.73m(2), respectively). The effect of admission eGFR on the primary outcome of death or major disability at 90 days (defined as modified Rankin Scale scores of 3-6) was analyzed using a multivariable logistic regression model. Potential effect modification of intensive BP lowering treatment by admission eGFR was assessed by interaction terms. Of 2,623 included participants, 912 (35%) and 280 (11%) had mildly and moderately/severely decreased eGFRs, respectively. Patients with moderately/severely decreased eGFRs had the greatest risk for death or major disability at 90 days (adjusted OR, 1.82; 95% CI, 1.28-2.61). Effects of early intensive BP lowering were consistent across different eGFRs (P=0.5 for homogeneity). Generalizability issues arising from a clinical trial population. Decreased eGFR predicts poor outcome in acute ICH. Early intensive BP lowering provides similar treatment effects in patients with ICH with decreased eGFRs.


Toxicity of 2,4-diacetylphloroglucinol (DAPG) to plant-parasitic and bacterial-feeding nematodes.
The antibiotic 2,4-diacetylphloroglucinol (DAPG) is produced by some isolates of the beneficial bacterium Pseudomonas fluorescens. DAPG is toxic to many organisms, and crop yield increases have been reported after application of DAPG-producing P. fluorescens. This study was conducted to determine whether DAPG is toxic to selected nematodes. The plant-parasitic nematodes Heterodera glycines, Meloidogyne incognita, Pratylenchus scribneri and Xiphinema americanum, and the bacterial-feeding nematodes Caenorhabditis elegans, Pristionchus pacificus, and Rhabditis rainai, were immersed in concentrations ranging from 0 to 100 μg/ml DAPG. Egg hatch and viability of juveniles and adults were determined. DAPG was toxic to X. americanum adults, with an LD₅₀ of 8.3 μg/ml DAPG. DAPG decreased M. incognita egg hatch, but stimulated C. elegans hatch during the first hours of incubation. Viability of M. incognita J2 and of C. elegans J1 and adults was not affected. There were no observed effects on the other nematodes. The study indicated that DAPG is not toxic to all nematodes, and did not affect the tested species of beneficial bacterial-feeding nematodes. Augmentation of DAPG-producing P. fluorescens populations for nematode biocontrol could be targeted to specific nematode species known to be affected by this compound and by other antibiotics produced by the bacteria, or these bacteria could be used for other possible effects, such as induced plant resistance.


Contribution of Contraceptive Discontinuation to Unintended Births in 36 Developing Countries.
High contraceptive discontinuation results in millions of women having an unmet need for contraception. However, its contribution to unintended fertility is not known. Reproductive calendar data in Demographic and Health Surveys in 36 countries are used to estimate the percent of unintended recent births attributable to contraceptive discontinuation. Contraceptive discontinuation accounted for about one-third of unintended recent births in all countries together. Method failure and contraception discontinued for other reasons accounted for most of this contribution. The contribution of contraceptive discontinuation to unintended births increases with the use of modern methods but decreases as method composition at a given level of contraceptive prevalence shifts toward methods with higher effectiveness and longer continuation. High contraceptive discontinuation in the past without changes in fertility intentions has resulted in millions of unintended births. This contribution is likely to increase with the anticipated increase in the use of modern methods. Enabling current users to reduce method failure and encouraging them to switch to another method after discontinuing the use of the original method will be an effective strategy to reduce contraceptive discontinuation and its contribution to unintended births.


Cost-Effectiveness of Antiviral Treatments for Pandemics and Outbreaks of Respiratory Illnesses, Including COVID-19: A Systematic Review of Published Economic Evaluations.
To review published economic evaluations of antiviral treatment for pandemics and outbreaks of respiratory illnesses. We conducted a systematic review to identify economic evaluations of antiviral treatment for pandemics and outbreaks of respiratory illnesses, including coronavirus disease 2019 (COVID-19). We searched Medline (EBSCOhost), EMBASE (Ovid), EconLit (Ovid), National Health Service Economic Evaluation Database (Ovid), and Health Technology Assessment (Ovid). The search was last rerun on July 5, 2020. Citation tracking and reference checking were used. Only full economic evaluations published as peer-reviewed articles in the last 10 years were included. Studies were quality assessed using the National Institute for Health and Care Excellence economic evaluation checklist. Overall, 782 records were identified, of which 14 studies met the inclusion criteria. The studies were mostly conducted in high-income countries. All were model-based. Seven (50%) were cost-utility analyses, 4 (28.6%) were cost-effectiveness analyses, 2 (14.3%) were cost-consequences analyses, and 1 (7.1%) was a cost-benefit analysis. Strategies including antiviral treatment were found to be either cost-saving or cost-effective, at the study-specific willingness-to-pay thresholds. Empirical treatment was more cost-effective than test-guided treatment for young adults but less so for older adults. Antiviral treatment for managing pandemics and outbreaks of respiratory illnesses that have very high case fatality rate, similar to COVID-19 pandemic, are likely to be cost-effective either as a standalone intervention or part of a multifaceted strategy. Investing in the development of such curative treatments and promptly evaluating their cost-effectiveness, relative to other strategies in use at the time of their introduction should be the focus going forward to inform resource allocation decisions particularly in low- and middle-income countries.


Upper limb entheseal change with the transition to agriculture in the Southeastern United States: a view from Moundville and the central Tombigbee River valley.
We analyzed entheseal change in 159 skeletons from Moundville and surrounding settlements using primary fibrocartilaginous attachments of the upper limbs. Risk of entheseal change did not differ bilaterally, suggesting a wide variety of activities were used to exploit a diverse ecosystem. Consistent with predictions, Mississippian (1000-1500 CE) agriculturalists experienced greater risk of entheseal change than did Late Woodland (500-900 CE) hunter-gatherers. Attachments used in arm flexion were most affected, while rotator cuff entheses remained consistent over time. A temporal increase in muscular changes in males in concert with faunal evidence for resurgence of larger game (e.g., deer) is consistent with continued reliance on hunting alongside domestication of maize. Among Mississippians, younger males appear to have been carrying out the most strenuous tasks, contrary to earlier studies that suggested a decline in male activities with domestication. Mound centers consistently experienced the greatest upper body changes, in spite of faunal and botanical data supporting provisioning of elites by outlying sites. Center males, respectively, experienced more than 26 and 12 times greater changes at elbow extensors and brachialis than those from outlying settlements, with a significant decline from young to middle age among adults. Center females experienced increased risk at biceps insertions and common extensors of the humeri - trends that disappeared with age. Overall findings suggest increased upper body demands and shifting sex and age-dependent divisions of labor with maize intensification, but trends across settlements point to significant status-related body size selection in center males, with fewer differences among females.


Radial head reconstruction in elbow fracture-dislocation: monopolar or bipolar prosthesis?
Monopolar and bipolar radial head prosthetic arthroplasties have been used successfully to treat elbow fracture-dislocation with unsalvageable radial head fractures. The relative stability of these two designs in different clinical situations is a topic of ongoing investigation. We tested the effects of monopolar and bipolar fixed-neck prosthetic radial head implants on improvement in elbow coronal and axial plane laxity in a terrible triad biomechanical model that accounted for lateral collateral ligament integrity and the presence of a transverse coronoid fracture. Kinematic data were collected on six fresh-frozen cadaveric upper extremities tested with passive motion throughout the flexion arc. Varus and valgus gravity stress were applied with the wrist in neutral position. A lateral collateral ligament reconstruction was simulated. We assessed instability after radial head resection and reconstruction with either a monopolar or bipolar implant in the presence of a transversely fractured (Regan and Morrey Type 2) or fixed coronoid process. With collateral ligament integrity, no difference was detected, with the numbers available, in valgus laxity between implants under valgus stress (p = 1.0). Laxity improvement with each prosthesis was higher when the coronoid was fractured (mean ± SD: monopolar: 7.4° ± 1.6°, p < 0.001; bipolar: 6.4° ± 1.6°, p = 0.003) than when it was fixed (monopolar: 4.0° ± 1.6°, p = 0.02; bipolar: 4.2° ± 1.6°, p = 0.01). With the numbers available, there was no difference in external rotation laxity between implants under valgus stress (p = 1.0). The greatest stabilizing effect of the prostheses occurred when the coronoid was fractured (monopolar: 3.3° ± 1.2°, p = 0.15; bipolar: 3.3° ± 1.2°, p = 0.17). Radial head arthroplasty offered no substantial stability under varus stress for varus or internal rotation laxity. In our terrible triad cadaveric model, coronoid fixation was effective in improving varus laxity with a monopolar or bipolar prosthesis in place. Also, both types of prostheses were effective in improving valgus and external rotation laxity to the elbow, regardless of coronoid status. With collateral ligaments reconstructed, no large kinematic differences were noted between implants regardless of the varus-valgus position or whether the coronoid was fractured or fixed. The data from our cadaveric model support the use of either implant type in terrible triad injuries if the collateral ligaments are intact or reconstructed.


Adsorption and Unfolding of Lysozyme at a Polarized Aqueous-Organic Liquid Interface.
The adsorption of proteins at the interface between two immiscible electrolyte solutions has been found to be key to their bioelectroactivity at such interfaces. Combined with interfacial complexation of organic phase anions by cationic proteins, this adsorption process may be exploited to achieve nanomolar protein detection. In this study, replica exchange molecular dynamics simulations have been performed to elucidate for the first time the molecular mechanism of adsorption and subsequent unfolding of hen egg white lysozyme at low pH at a polarized 1,2-dichloroethane/water interface. The unfolding of lysozyme was observed to occur as soon as it reaches the organic-aqueous interface, which resulted in a number of distinct orientations at the interface. In all cases, lysozyme interacted with the organic phase through regions rich in nonpolar amino acids, such that the side chains are directed toward the organic phase, whereas charged and polar residues were oriented toward the aqueous phase. By contrast, as expected, lysozyme in neat water at low pH does not exhibit significant structural changes. These findings demonstrate the key influence of the organic phase upon adsorption of lysozyme under the influence of an electric field, which results in the unfolding of its structure.


Orientation dependence in homologous recombination.
Homologous recombination was investigated in Escherichia coli with two plasmids, each carrying the homologous region (two defective neo genes, one with an amino-end deletion and the other with a carboxyl-end deletion) in either direct or inverted orientation. Recombination efficiency was measured in recBC sbcBC and recBC sbcA strains in three ways. First, we measured the frequency of cells carrying neo+ recombinant plasmids in stationary phase. Recombination between direct repeats was much more frequent than between inverted repeats in the recBC sbcBC strain but was equally frequent in the two substrates in the recBC sbcA strain. Second, the fluctuation test was used to exclude bias by a rate difference between the recombinant and parental plasmids and led to the same conclusion. Third, direct selection for recombinants just after transformation with or without substrate double-strand breaks yielded essentially the same results. Double-strand breaks elevated recombination in both the strains and in both substrates. These results are consistent with our previous findings that the major route of recombination in recBC sbcBC strains generates only one recombinant DNA from two DNAs and in recBC sbcA strains generates two recombinant DNAs from two DNAs.


Identification and characterization of a functional, alternatively spliced Toll-like receptor 7 (TLR7) and genomic disruption of TLR8 in chickens.
Based upon the recognition of antiviral compounds and single stranded viral RNA the Toll-like receptors TLR7 and TLR8 are suggested to play a significant role in initiating antiviral immune responses. Here we report the molecular characterization of the chicken TLR7/8 loci which revealed an intact TLR7 gene and fragments of a TLR8-like gene with a 6-kilobase insertion containing chicken repeat 1 (CR1) retroviral-like insertion elements. The chicken TLR7 gene encodes a 1047-amino-acid protein with 62% identity to human TLR7 and a conserved pattern of predicted leucine-rich repeats. Highest levels of chicken TLR7 mRNA were detected in immune-related tissues and cells, especially the spleen, caecal, tonsil and splenic B cells. Alternative spliced forms of TLR7 mRNA were identified in chicken, mouse and human and expressed in similar tissues and cell types to the major form of chicken TLR7. The chicken TLR7+ HD11 cell line and fresh splenocytes produced elevated levels of interleukin-1beta (IL-1beta) mRNA after exposure to the agonists R848 and loxoribine. Interestingly, none of the TLR7 agonists stimulated increased type I interferon (IFN) mRNA whereas poly(I:C) (a TLR3 agonist) up-regulated both chicken IFN-alpha and chicken IFN-beta mRNA. In contrast, TLR7 agonists, particularly R848 and poly(U) stimulated up-regulation of chicken IL-1beta, and chicken IL-8 mRNAs more effectively than poly(I:C). Stimulation of chicken TLR7 with R848 was chloroquine sensitive, suggesting signalling within an endosomal compartment, as for mammalian TLR7. The deletion of TLR8 in galliforms, accompanied with the differential response after exposure to TLR7 agonists, offers insight into the evolution of vertebrate TLR function.


Novel R-roscovitine NO-donor hybrid compounds as potential pro-resolution of inflammation agents.
Neutrophils play a pivotal role in the pathophysiology of multiple human inflammatory diseases. Novel pharmacological strategies which drive neutrophils to undergo programmed cell death (apoptosis) have been shown to facilitate the resolution of inflammation. Both the cyclin-dependent kinase inhibitor (CDKi) R-roscovitine and nitric oxide (NO) have been shown to enhance apoptosis of neutrophils and possess pro-resolution of inflammation properties. In order to search for new multi-target pro-resolution derivatives, here we describe the design, synthesis and investigation of the biological potential of a small series of hybrid compounds obtained by conjugating R-roscovitine with two different NO-donor moieties (compounds 2, 9a, 9c). The synthesized compounds were tested as potential pro-resolution agents, with their ability to promote human neutrophil apoptosis evaluated. Both compound 9a and 9c showed an increased pro-apoptotic activity when compared with either R-roscovitine or structurally related compounds devoid of the ability to release NO (des-NO analogues). Inhibition of either NO-synthase or soluble guanylate cyclase did not affect the induction of apoptosis by the R-roscovitine derivatives, similar to that reported for other classes of NO-donors. In contrast the NO scavenger PTIO prevented the enhanced apoptosis seen with compound 9a over R-roscovitine. These data show that novel compounds such as CDKi-NO-donor hybrids may have additive pro-resolution of inflammation effects.


Association between intrapartum interventions and breastfeeding duration.
Few women who reside in Hong Kong exclusively breastfeed, and one-half stop breastfeeding within the first few months. There is little research in this population on the association between intrapartum interventions and breastfeeding duration. A sample of 1280 mother-infant pairs were recruited from the obstetric units of 4 public hospitals in Hong Kong and followed prospectively for 12 months or until the infant was weaned. The outcome variables for this analysis were the duration of any and exclusive breastfeeding. Predictor variables were 4 intrapartum interventions: receipt of opioid pain medication, induction versus spontaneous labor, epidural administration, and mode of birth. We used Cox proportional hazards modeling to assess the impact of intrapartum interventions on the duration of any and exclusive breastfeeding, and we constructed Kaplan-Meier survival curves to evaluate the cumulative impact of multiple intrapartum interventions on breastfeeding outcomes. Bivariate analysis showed that induction of labor (hazard ratio [HR], 1.24; 95% confidence interval [CI], 1.09-1.41), opioid pain medication (HR, 1.21; 95% CI, 1.06-1.37), and having an emergency cesarean birth (HR, 1.22; 95% CI, 1.01-1.48) were associated with a shorter duration of any breastfeeding. Induction of labor (HR, 1.23; 95% CI, 1.08-1.39) and having an emergency cesarean birth (HR, 1.25; 95% CI, 1.05-1.51) were associated with a shorter duration of exclusive breastfeeding. After controlling for known confounding variables, there was no longer any association between individual intrapartum interventions and the duration of any or exclusive breastfeeding. The median duration of breastfeeding for participants who experienced a natural birth with no intrapartum interventions was 9 weeks compared with 5 weeks for participants who experienced at least 3 intrapartum interventions. Clinicians working with new breastfeeding mothers should focus on providing additional support to mothers who experience a difficult labor and birth with multiple interventions to improve their breastfeeding experiences.


Idiopathic Acquired Temporal Wedge Visual Field Defects.
Our aim is to report 13 unusual cases of acquired, temporal sectoral scotomas. Such stationary "wedge" field defects have been reported previously in cases of presumed congenital nasal hypoplasia of the optic disc and as a complication of vitreoretinal surgery. To our knowledge, the literature provides no reports of similar defects occurring spontaneously. This is a descriptive clinical case series of 13 patients presenting with sub-acute monocular temporal visual field loss. All were clinically assessed and investigated with Goldmann perimetry, automated Humphrey visual fields, retinal optical coherence tomography, orbital ultrasound, and standard and multi-focal electroretinography. Cases were followed with serial perimetry for a mean of 3.9 years (range: 6 months to 10 years). Goldmann and Humphrey perimetry both demonstrated "wedge"-shaped defects extending temporally from an apex contiguous with, or lateral to, the blind spot. There was no evidence of optic disc drusen, glaucoma, disc hypoplasia, or focal retinitis. Sectoral optic disc swelling was not present in any patient at presentation. In all cases, the visual field defect remained stable. One patient developed a similar defect in the fellow eye after an interval of 5 years. Here we describe 13 cases of acquired, stationary temporal wedge scotomas, novel in the literature. Although the aetiology is uncertain, we propose damage to the nasal rim of the optic disc as a likely mechanism.


Context-dependent interactions and the regulation of species richness in freshwater fish.
Species richness is regulated by a complex network of scale-dependent processes. This complexity can obscure the influence of limiting species interactions, making it difficult to determine if abiotic or biotic drivers are more predominant regulators of richness. Using integrative modeling of freshwater fish richness from 721 lakes along an 11o latitudinal gradient, we find negative interactions to be a relatively minor independent predictor of species richness in lakes despite the widespread presence of predators. Instead, interaction effects, when detectable among major functional groups and 231 species pairs, were strong, often positive, but contextually dependent on environment. These results are consistent with the idea that negative interactions internally structure lake communities but do not consistently 'scale-up' to regulate richness independently of the environment. The importance of environment for interaction outcomes and its role in the regulation of species richness highlights the potential sensitivity of fish communities to the environmental changes affecting lakes globally.


Oxidative Stress Parameters and Erythrocyte Membrane Adenosine Triphosphatase Activities in Streptozotocin-induced Diabetic Rats Administered Aqueous Preparation of Kalanchoe Pinnata Leaves.
Diabetes mellitus is a chronic metabolic disease that according to the World Health Organization affects more than 382 million people. The rise in diabetes mellitus coupled with the lack of an effective treatment has led many to investigate medicinal plants to identify a viable alternative. To evaluate red blood cell (RBC) membrane adenosine triphosphatase (ATPase) activities and antioxidant levels in streptozotocin-induced diabetic rats administered aqueous preparation of Kalanchoe pinnata leaves. Diabetes mellitus was induced in rats by a single administration of streptozotocin (60 mg/kg). Diabetic rats were then treated with aqueous K. pinnata preparation (three mature leaves ~ 9.96 g/70 kg body weight or about 0.14 g/kg body weight/day) for 30 days. Serum glucose, RBC membrane ATPase activities, and antioxidant levels were determined. We noted weight loss and reduced food consumption in the treated diabetic group. Serum glucose levels were reduced in the treated diabetic group compared to the other groups. Superoxide dismutase activity and glutathione levels were not significantly elevated in the treated group compared to the diabetic group. However, serum catalase activity was significantly (P < 0.05) increased in the treated diabetic group compared to the other groups. Serum thiobarbituric acid reactive substances were not significantly altered among the groups. There was a significant (P < 0.05) increase in Mg(2+) ATPase activity and a nonsignificant increase in Na(+)/K(+) ATPase activity in the RBC membrane of the treated diabetic group compared to the diabetic group. The consumption of aqueous preparation of K. pinnata may accrue benefits in the management of diabetes by lowering oxidative stress often associated with the disease and improving the availability of cellular magnesium through an increase in the magnesium ATPase pump in the RBC membrane for increased cellular metabolism of glucose through the glycolytic pathway. We noted weight loss and reduced food consumption in the diabetic rats treated with K. pinnata preparationSerum glucose levels were reduced in diabetic rats treated with K. pinnata preparationSerum catalase activity was significantly (P < 0.05) increased in diabetic rats treated with K. pinnata preparationWe also noted a significant (P < 0.05) increase in Mg(2+) ATPase activity in the RBC membranes of diabetic rats treated with K. pinnata preparationOverall, the consumption of aqueous preparation of K. pinnata lowered oxidative stress often associated with diabetes and improved availability of cellular magnesium through an increase in magnesium ATPase pump in the RBC membrane.


Lanthanide single molecule magnets: progress and perspective.
The last few years have seen a huge renaissance in the study of single molecule magnets (SMMs) thanks to the extensive applications of lanthanide ions with large inherent anisotropy in molecular magnetism. Particularly, the recent theoretical developments and the experimental expansion into the organometallic avenue have led to an eye-catching boost in this field. Here we highlight the recent progress in this fascinating and challenging field, with emphasis on several combined experimental and theoretical studies.


Evaluation of insulin-like growth factor-1, total thyroxine, feline pancreas-specific lipase and urinary corticoid-to-creatinine ratio in cats with diabetes mellitus in Switzerland and the Netherlands.
Objectives The aim of the study was to evaluate circulating insulin-like growth factor-1 (IGF-1), feline pancreas-specific lipase (fPLI) and total thyroxine (TT4) concentrations and urinary corticoid-to-creatinine ratio (UCCR) as indicators for the prevalence of acromegaly, pancreatitis, hyperthyroidism and hypercortisolism in cats with diabetes mellitus. Methods Blood and urine samples were collected from diabetic cats treated in primary care clinics in Switzerland and the Netherlands. Standardised questionnaires and physical examination forms provided clinical information from owners and veterinarians. Laboratory testing included serum biochemistry profile analysis and measurement of circulating fructosamine, IGF-1, fPLI, and TT4 concentrations and UCCR. CT of the pituitary gland was performed using a multidetector computed tomography scanner. Results Blood samples were available from 215 cats and urine samples were collected at home from 117 cats. Age ranged from 2-18 years (median 12 years) and body weight from 2.7-12.3 kg (median 5.5 kg). Sixty-five percent of the cats were castrated male and 35% were female (33% spayed); 82% were domestic shorthair cats. Eighty percent of cats received a porcine insulin zinc suspension, 19.5% insulin glargine and 0.5% a human neutral protamine hagedorn insulin. Thirty-six of 202 (17.8%) cats had IGF-1 concentrations >1000 ng/ml. Serum fPLI, and TT4 concentrations and UCCR were increased in 86/196 (43.9%), 9/201 (4.5%) and 18/117 cats (15.3%), respectively. Prevalence did not differ between countries. Conclusions Hyperthyroidism is rare, whereas increased fPLI concentration, possibly reflecting pancreatitis, is common in diabetic cats. The high UCCR may reflect activation of the hypothalamus-pituitary-adrenal axis, which also occurs in diabetic humans. The percentage of cats with increased IGF-1 was high but lower than reported in recent studies.


Tcf19 is a novel islet factor necessary for proliferation and survival in the INS-1 β-cell line.
Recently, a novel type 1 diabetes association locus was identified at human chromosome 6p31.3, and transcription factor 19 (TCF19) is a likely causal gene. Little is known about Tcf19, and we now show that it plays a role in both proliferation and apoptosis in insulinoma cells. Tcf19 is expressed in mouse and human islets, with increasing mRNA expression in nondiabetic obesity. The expression of Tcf19 is correlated with β-cell mass expansion, suggesting that it may be a transcriptional regulator of β-cell mass. Increasing proliferation and decreasing apoptotic cell death are two strategies to increase pancreatic β-cell mass and prevent or delay diabetes. siRNA-mediated knockdown of Tcf19 in the INS-1 insulinoma cell line, a β-cell model, results in a decrease in proliferation and an increase in apoptosis. There was a significant reduction in the expression of numerous cell cycle genes from the late G1 phase through the M phase, and cells were arrested at the G1/S checkpoint. We also observed increased apoptosis and susceptibility to endoplasmic reticulum (ER) stress after Tcf19 knockdown. There was a reduction in expression of genes important for the maintenance of ER homeostasis (Bip, p58(IPK), Edem1, and calreticulin) and an increase in proapoptotic genes (Bim, Bid, Nix, Gadd34, and Pdia2). Therefore, Tcf19 is necessary for both proliferation and survival and is a novel regulator of these pathways.


An interesting case of systemic lupus erythematosus presenting with hypercalcemia: A diagnostic dilemma.
Hypercalcemia is common in primary hyperparathyroidism malignancies and even in tuberculosis. Interestingly, systemic lupus erythematosus (SLE) rarely presents with hypercalcemia. We describe an interesting case of SLE in a patient who was otherwise thought to have either tuberculosis or a malignancy. The patient initially presented with feeling unwell, with generalized lymphadenopathy, bilateral pleural effusion, and bilateral corneal calcium deposits secondary to severe hypercalcemia. The diagnosis of SLE was made based on positivity of antinuclear antibodies (ANA) and anti-dsDNA, the presence of serositis, lymphadenopathy, autoimmune hemolytic anemia, and constitutional symptoms. She was treated with steroids, with tremendous improvement in her general well-being, resolution of lymphadenopathy and pleural effusion, and normalization of her hemoglobin and serum calcium. The atypical presentation of SLE with hypercalcemia with pleural effusion is discussed. SLE should be one of the differential diagnoses in patients presenting with severe hypercalcemia.


Assessment of the frequency of routine removal of dental plaque prior to caries diagnosis by dentists in three cities in southern Brazil.
The aim of this study was to assess the frequency of routine use of dental prophylaxis prior to visual inspection, in order to diagnose caries, by dentists with different lapses of time after graduating time. One hundred and fifty one Brazilian dentists were interviewed in 3 Brazilian cities to determine if they usually remove dental plaque prior to visual inspection for caries diagnosis. The dentists were stratified according to year of graduation. The association between the lapse of time after graduating and the practice of routinely removing dental plaque before clinical examination was tested using the chi-square test with a significance level of 5%. Only 28.5% of the dentists reported that they usually remove dental plaque prior to clinical examination. The dentists who graduated in the last 15 years presented the lowest percentages of plaque removal prior to clinical examination (15.1%), whereas the more experienced dentists reported that they perform prophylaxis more frequently. Of the professionals who graduated from 1960-1975, 23.9% reported that they performed dental plaque removal prior to diagnosis, whereas the figure for those graduating from 1976-1990 was 46.2%. Most of the dentists interviewed reported that they did not remove dental plaque prior to performing visual diagnosis of caries.


Phase 2 study of eribulin in patients with previously treated advanced or metastatic soft tissue sarcoma.
Eribulin, a microtubule dynamics inhibitor, is approved for the treatment of patients with breast cancer and soft tissue sarcoma. We investigated the efficacy and safety of eribulin in Japanese patients with soft tissue sarcoma. This open-label, multicenter, nonrandomized, Phase 2 study enrolled Japanese patients with measurable, advanced/metastatic soft tissue sarcoma of high/intermediate grade and ≥1 prior chemotherapy for advanced disease. Patients received eribulin mesilate 1.4 mg/m2 intravenously over 2–5 minutes on Days 1 and 8 of a 21-day cycle. The primary endpoint was progression-free rate at 12 weeks. Secondary endpoints included overall survival, progression-free survival and safety. Efficacy analyses were stratified by histology (liposarcoma or leiomyosarcoma, and other subtypes). Overall, 52 patients were enrolled and 51 patients were treated. Patients with liposarcoma/leiomyosarcoma (n = 35) had similar characteristics to those with other subtypes (n = 16), except for a higher proportion of women (63% vs 38%, respectively) and patients with Eastern Cooperative Oncology Group performance status 0 (57% vs 44%). Progression-free rate at 12 weeks was 60% in liposarcoma/leiomyosarcoma patients, 31% in other subtypes and 51% overall. Median progression-free survival was 5.5 months in liposarcoma/leiomyosarcoma patients, 2.0 months in other subtypes and 4.1 months overall. Median overall survival was 17.0 months in liposarcoma/leiomyosarcoma patients, 7.6 months in other subtypes and 13.2 months overall. The most common Grade 3–4 adverse events were neutropenia (86%), leukopenia (75%), lymphopenia (33%), anemia (14%) and febrile neutropenia (8%). Eribulin showed clinical activity with a manageable safety profile in previously treated Japanese patients with advanced/metastatic soft tissue sarcoma.


Differential gene expression responses distinguish contact and respiratory sensitizers and nonsensitizing irritants in the local lymph node assay.
Genomic approaches have the potential to enhance the specificity and predictive accuracy of existing toxicology endpoints, including those for chemical sensitization. The present study was conducted to determine whether gene expression responses can distinguish contact sensitizers (1-chloro-2,4-dinitrobenzene [DNCB] and hexyl cinnamic aldehyde [HCA]), respiratory sensitizers (ortho-phthalaldehyde and trimellitic anhydride [TMA]), and nonsensitizing irritants (methyl salicylate [MS] and nonanoic acid [NA]) in the local lymph node assay (LLNA). Female Balb/c mice received doses of each chemical as per the standard LLNA dosing regimen on days 1, 2, and 3. Auricular lymph nodes were analyzed for tritiated thymidine ((3)HTdR) incorporation on day 6 and for gene expression responses on days 6 and 10. All chemicals induced dose-dependent increases in stimulation index, which correlated strongly with the number of differentially expressed genes. A majority of genes modulated by the irritants were similarly altered by the sensitizers, consistent with the irritating effects of the sensitizers. However, a select number of responses involved with immune-specific functions, such as dendritic cell activation, were unique to the sensitizers and may offer the ability to distinguish sensitizers from irritants. Genes for the mast cell proteases 1 and 8, Lgals7, Tim2, Aicda, Il4, and Akr1c18 were more strongly regulated by respiratory sensitizers compared with contact sensitizers and may represent potential biomarkers for discriminating between contact and respiratory sensitizers. Collectively, these data suggest that gene expression responses may serve as useful biomarkers to distinguish between respiratory and contact sensitizers and nonsensitizing irritants in the LLNA.


L-Carnitine Supplementation Increases Trimethylamine-N-Oxide but not Markers of Atherosclerosis in Healthy Aged Women.
L-carnitine can be metabolized to trimethylamine N-oxide (TMAO), a molecule that promotes atherogenesis through its interaction with macrophages and lipid metabolism. The aim of the present study was to assess whether L-carnitine supplementation may promote changes in selected serum biomarkers of atherosclerosis. Before the start, in the mid-point and after completing the 24-weeks supplementation protocol, fasting blood samples were taken from the antecubital vein. Plasma free L-carnitine and TMAO were determined by the UPLC/MS/MS method. Serum proteins were determined by the enzyme immunoassay method using commercially available kits. Total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, and triglycerides have been determined using standard automatic analyzer. L-carnitine supplementation elevated fasting plasma carnitine in the mid-point of our study and it remained increased until the end of supplementation period. Moreover, it induced tenfold increase in plasma TMAO concentration but did not affect serum C-reactive protein, interleukin-6, tumour necrosis factor-α, L-selectin, P-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1 or lipid profile markers. We demonstrated that -although oral L-carnitine supplementation significantly -increased plasma TMAO concentration, no lipid profile changes or other markers of adverse cardiovascular events were detected in healthy aged women over the period of 24 weeks.


Polyphenols-rich Vernonia amygdalina shows anti-diabetic effects in streptozotocin-induced diabetic rats.
This study aims to investigate the hypoglycemic properties of Vernonia amygdalina Del. (VA) and its possible mechanisms of action in a single-dose STZ induced diabetic rat model. A dose-response study was conducted to determine optimum dose for the hypoglycemic effect of VA in STZ-induced diabetic rats. The optimum dose (400 mg/kg) was used throughout the 28-day chronic study. Body weight, food and water intakes of the rats were monitored daily. Fasting blood serum, pancreas, liver and soleus muscle were collected for biochemical analyses. Chemical composition of VA was analysed using HPLC and LC-ESI-MS. The study reveals that ethanolic extract of VA contains high level of polyphenols mainly 1,5-dicaffeoyl-quinic acid, dicaffeoyl-quinic acid, chlorogenic acid and luteolin-7-O-glucoside. In an oral glucose tolerance test, 400 mg/kg VA exhibited a significant improvement in glucose tolerance of the STZ-induced diabetic rats. 28-day treatment with 400 mg/kg VA resulted in 32.1% decrease in fasting blood glucose compared to diabetic control. VA also caused significant decrease (18.2% and 41%) in triglyceride and total cholesterol level. Besides, VA showed protective effect over pancreatic β-cells against STZ-induced damage, causing a slight increase in insulin level compared to diabetic control. VA administration also showed positive regulation of the antioxidant system, both enzymatic and non-enzymatic. Furthermore, VA was found to increase expression of GLUT 4 (24%) in rat skeletal muscle. Further tissue fractionation revealed that it can increase the GLUT 4 translocation (35.7%) to plasma membrane as well, suggesting that VA may stimulate skeletal muscle's glucose uptake. This observation is in line with the restoration in skeletal muscle glycogenesis of VA-treated group. However, no alteration was observed in GLUT 1 expression. In addition, VA also suppressed (40% inhibition) one of the key hepatic gluconeogenic enzymes, glucose-6-phosphatase (G6Pase). VA possesses antihyperglycemic effect, most probably through increasing GLUT 4 translocation and inhibiting hepatic G6Pase. The polyphenols in the extract may be the candidates that are responsible for the above-mentioned biological activities.


Early Childhood Caries in Indigenous Communities.
The oral health of Indigenous children of Canada (First Nations, Inuit, and Métis) and the United States (American Indian and Alaska native) is a major child health disparity when compared with the general population of both countries. Early childhood caries (ECC) occurs in Indigenous children at an earlier age, with a higher prevalence, and at much greater severity than in the general population. ECC results in adverse oral health, affecting childhood health and well-being, and may result in high rates of costly surgical treatment under general anesthesia. ECC is an infectious disease that is influenced by multiple factors, but the social determinants of health are particularly important. This policy statement includes recommendations for preventive and clinical oral health care for infants, toddlers, preschool-aged children, and pregnant women by primary health care providers. It also addresses community-based health-promotion initiatives and access to dental care for Indigenous children. This policy statement encourages oral health interventions at early ages in Indigenous children, including referral to dental care for the use of sealants, interim therapeutic restorations, and silver diamine fluoride. Further community-based research on the microbiology, epidemiology, prevention, and management of ECC in Indigenous communities is also needed to reduce the dismally high rate of caries in this population.


Fluorescent nanoparticles in the popular pizza: properties, biodistribution and cytotoxicity.
Food-borne nanoparticles that are generated during the thermal processing of various consumed foods are of great concern due to their unique properties. In this study, the presence of fluorescent nanoparticles (FNPs) in pizza, their biodistribution and cytotoxicity were investigated. The spherical FNPs have a diameter of about 3.33 nm. X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy analysis revealed that they contained 68.21% C, 27.44% O, 2.75% N and 1.60% S, and the functional groups on their surface included -OH, -COOH, C[double bond, length as m-dash]C, -NH2 and C[double bond, length as m-dash]O. In vitro and in vivo biodistribution of pizza FNPs was evaluated using normal rat kidney (NRK) cells, onion epidermal cells, Caenorhabditis elegans and mice. The fluorescence microscopy images clearly indicate that the pizza FNPs appear to be localized within the cytoplasm. However, the FNPs remained restricted to the extracellular space of the onion epithelium and did not enter the onion cell cytoplasm because of the cell wall. The FNPs were swallowed by the Caenorhabditis elegans worms when exposed to food OP50 and distributed within the pharynx, intestine and anus. Obvious fluorescence of the FNPs in the stomach, intestine, liver, lung and kidney was observed for the FNPs in mouse organs, but not the brain, heart, and spleen. Furthermore, the produced FNPs were found to cause cell cycle arrest at the G0/G1 phase in NRK cells, and resulted in cell apoptosis at high doses. The outcome of this research offers an important insight into the nature of thermal processing-induced nanoparticles and their in vivo and in vitro biological effects.


Mechanomyography-Based Wearable Monitor of Quasi-Isometric Muscle Fatigue for Motor Neural Prostheses.
A motor neural prosthesis based on surface functional electrical stimulation (sFES) can restore functional movement (e.g., standing, walking) in patients with a spinal cord injury (SCI). sFES generates muscle contractions in antigravity muscles and allows balance-assisted standing. This induced standing has several benefits, such as improved cardiovascular function, decreased incidence of urinary infections, reduced joint contractures, and muscle atrophy. The duration of sFES assisted standing is limited due to the quick onset of muscle fatigue. Currently, there is no method available to reliably estimate real-time muscle fatigue during sFES. Simply monitoring the M-wave changes is not suitable due to the high signal disturbances that arise during multi-channel electrical stimulation. Mechanomyography (MMG) is immune to electrical stimulation artifacts and can be used to detect subtle vibrations on the surface of the skin related to activation of the underlying muscle's motor units (MU). The aim of this study was to develop a method for detecting muscle fatigue brought on by sFES. The method was tested in three different heads of the quadriceps muscle in SCI patients during electrically elicited quasi-isometric contraction. Six spinal cord-injured male volunteers, with no voluntary control of the quadriceps muscle participated in the study. Electrical bursts of voltage-controlled monophasic square pulses at frequencies of 1 kHz (50% duty cycle) at 50 Hz (15% duty cycle) were used to generate thigh muscle contractions that controlled the knee joint in the sagittal plane. The pulse amplitudes were set to position the knee joint at a 5° angle from the horizontal plane and when the knee angle dropped to 20° (e.g., the quadriceps were unable to hold the lower leg in the desired position), the test was terminated. Two data segments lasting 10 s each, at the beginning and end of each test, were analyzed. The muscle contraction was assessed by MMG sensors positioned on the rectus femoris, vastus lateralis, and vastus medialis muscles. Data segments were decomposed into 11 frequency bands using a Cauchy wavelet transform. In the initial time interval (non-fatigued muscle), the power peak was concentrated in the 11.31 Hz frequency band. In the final interval (muscle fatigued) this peak shifted to lower frequencies (2 and 6 Hz frequency bands). The decreased frequency was most prominent during the last 4 s of the recordings. It was shown that MMG could be used as a real-time indicator of muscle fatigue during FES-induced isometric contraction of quadriceps; hence, MMG could be used in closed-loop control as a fatigue detector. Subsequent studies for non-isometric contractions could possibly lead to prediction of muscle fatigue before contractile failure during functional use of the muscle.


Factors contributing to food insecurity among women living with HIV in the Dominican Republic: A qualitative study.
Food insecurity contributes to poor health outcomes among people living with HIV. In Latin America and the Caribbean, structural factors such as poverty, stigma, and inequality disproportionately affect women and may fuel both the HIV epidemic and food insecurity. We examined factors contributing to food insecurity among women living with HIV (WLHIV) in the Dominican Republic (DR). Data collection included in-depth, semi-structured interviews in 2013 with 30 WLHIV with indications of food insecurity who resided in urban or peri-urban areas and were recruited from local HIV clinics. In-person interviews were conducted in Spanish. Transcripts were coded using content analysis methods and an inductive approach to identify principal and emergent themes. Respondents identified economic instability as the primary driver of food insecurity, precipitated by enacted stigma in the labor and social domains. Women described experiences of HIV-related labor discrimination in formal and informal sectors. Women commonly reported illegal HIV testing by employers, and subsequent dismissal if HIV-positive, especially in tourism and free trade zones. Enacted stigma in the social domain manifested as gossip and rejection by family, friends, and neighbors and physical, verbal, and sexual abuse by intimate partners, distancing women from sources of economic and food support. These experiences with discrimination and abuse contributed to internalized stigma among respondents who, as a result, were fearful and hesitant to disclose their HIV status; some participants reported leaving spouses and/or families, resulting in further isolation from economic resources, food and other support. A minority of participants described social support by friends, spouses, families and support groups, which helped to ameliorate food insecurity and emotional distress. Addressing food insecurity among WLHIV requires policy and programmatic interventions to enforce existing laws designed to protect the rights of people living with HIV, reduce HIV-related stigma, and improve gender equality.


Minimum Effective Doses of Succinylcholine and Rocuronium During Electroconvulsive Therapy: A Prospective, Randomized, Crossover Trial.
Neuromuscular blockade is required to control excessive muscle contractions during electroconvulsive therapy (ECT). In a crossover, assessor-blinded, prospective randomized study, we studied the minimum effective dose (MED) of succinylcholine and rocuronium for ECT. The MED was the lowest dose to provide a predefined qualitative measure of acceptable control of muscle strength during induced convulsions. Succinylcholine (0.8 mg kg) or rocuronium (0.4 mg kg) was randomly administered in 227 ECT sessions to 45 patients. The dose was incrementally increased or decreased by 10% based on 2 psychiatrists' (blinded to treatment) assessment of "acceptable" or "not acceptable" control of evoked muscle contractions (sufficient versus insufficient or excessive paralysis). The neuromuscular transmission was monitored quantitatively until full recovery. In our study, the MEDs of succinylcholine and rocuronium to produce acceptable ECT conditions in 50% of patients (MED50ECT) were 0.85 mg kg (95% confidence interval [CI], 0.77-0.94) and 0.41 mg kg (95% CI, 0.36-0.46) and in 90% of patients (MED90ECT) were 1.06 mg kg (95% CI, 1.0-1.27) and 0.57 mg kg (95% CI, 0.5-0.6), respectively. Nadir twitch height for acceptable muscle activity was 0% (0-4) and 4% (0-30; P < 0.001), respectively, and the time to recovery of the neuromuscular transmission was 9.7 ± 3.5 and 19.5 ± 5.7 minutes, respectively. A twitch suppression of >90% is needed for control of motor contractions during ECT. The initial ECT dose of succinylcholine should be selected based on each patient's preprocedural condition, ranging between 0.77 and 1.27 mg kg to produce acceptable muscle blockade in 50% to 90% of patients. Rocuronium-neostigmine combination is a safe alternative if appropriately dosed (0.36-0.6 mg kg) and monitored.


Sustained regression of tumors upon MYC inactivation requires p53 or thrombospondin-1 to reverse the angiogenic switch.
The targeted inactivation of oncogenes offers a rational therapeutic approach for the treatment of cancer. However, the therapeutic inactivation of a single oncogene has been associated with tumor recurrence. Therefore, it is necessary to develop strategies to override mechanisms of tumor escape from oncogene dependence. We report here that the targeted inactivation of MYC is sufficient to induce sustained regression of hematopoietic tumors in transgenic mice, except in tumors that had lost p53 function. p53 negative tumors were unable to be completely eliminated, as demonstrated by the kinetics of tumor cell elimination revealed by bioluminescence imaging. Histological examination revealed that upon MYC inactivation, the loss of p53 led to a deficiency in thrombospondin-1 (TSP-1) expression, a potent antiangiogenic protein, and the subsequent inability to shut off angiogenesis. Restoration of p53 expression in these tumors re-established TSP-1 expression. This permitted the suppression of angiogenesis and subsequent sustained tumor regression upon MYC inactivation. Similarly, the restoration of TSP-1 alone in p53 negative tumors resulted in the shut down of angiogenesis and led to sustained tumor regression upon MYC inactivation. Hence, the complete regression of tumor mass driven by inactivation of the MYC oncogene requires the p53-dependent induction of TSP-1 and the shut down of angiogenesis. Notably, overexpression of TSP-1 alone did not influence tumor growth. Therefore, the combined inactivation of oncogenes and angiogenesis may be a more clinically effective treatment of cancer. We conclude that angiogenesis is an essential component of oncogene addiction.


Urogynecology training and practice patterns after residency.
The perspective of recent graduates is important in assessing the adequacy of training and for improving clinical and surgical education. The objective of this study was to evaluate the urogynecology experience of recent Obstetrics and Gynecology residency graduates, to examine perceived and actual surgical load during and after residency, and to assess comfort level with diagnosing and treating urinary incontinence and pelvic organ prolapse. Questionnaire assessing satisfaction with urogynecology rotation, perceived surgical load in residency, estimates of surgical procedures in current practice, and comfort level with diagnosing and treating incontinence and prolapse. University of Texas Southwestern Medical Center, Dallas, Texas. Graduates from the University of Texas Southwestern Obstetrics and Gynecology residency program (1997-2006). Fifty-five percent (82/150) responded, with most being Caucasian, female, in private practice, and practicing in the Southern part of the United States. Forty-one of seventy-six (54%) rated their urogynecology experience as either acceptable, above average or superior. The most common procedures performed in residency were cystoscopy (mean, 11.3 ± 5.2 cases per resident) and anterior (mean, 8.9 ± 4.3 cases per resident) and posterior repair (mean, 11.1 ± 5.7 cases per resident). Increasing the surgical volume was reported as the single most important factor that would have enhanced their training. In practice, midurethral slings were the most commonly performed incontinence procedures. Few were performing other urogynecologic cases in practice, except for anterior-posterior colporrhaphy and cystoscopy. Sixty-six of 70 (94%) in practice were comfortable with diagnosing prolapse and incontinence. Seventy percent and 80% estimated that less than 1 quarter of their patient population had prolapse or incontinence, respectively. Recent graduates from this program are comfortable making the diagnosis of prolapse and incontinence. The most commonly performed surgical procedures in residency and in practice include anterior and posterior colporrhaphy and cystoscopy.


From word reading to multisentence comprehension: Improvements in brain activity in children with autism after reading intervention.
Children with ASD show a unique reading profile characterized by decoding abilities equivalent to verbal abilities, but with lower comprehension skills. Neuroimaging studies have found recruitment of regions primarily associated with visual processing (e.g., fusiform gyrus and medial parietal cortex), but reduced activation in frontal and temporal regions, when reading in adults with ASD. The purpose of this study was to assess neural changes associated with an intense reading intervention program in children with ASD using three fMRI tasks of reading. 25 children with ASD were randomly assigned to a treatment (ASD-EXP) or waitlist group (ASD-WLC). Children participated in a reading intervention program (4-hour sessions per day, 5 days a week for 10 weeks). We utilized three tasks: word, sentence, and multisentence processing, each with differential demands of reading comprehension. fMRI data were acquired at each of two scanning sessions 10-weeks apart. Across tasks, post-intervention results revealed that the ASD-EXP group showed greater activation in bilateral precentral gyrus and the postcentral gyrus, visual processing regions (e.g., occipital cortex, fusiform gyrus), and frontal regions. In the word task, left thalamus and the right angular gyrus (AG) activation was unique to the ASD-EXP group post-intervention. Sentence tasks showed differential activation of core language areas (e.g., IFG, IPL) post-intervention. Our results provide evidence for differential recruitment of brain regions based on task demands in children with ASD, and support the potential of targeted interventions to alter brain activation in response to positive gains in treatment. Children with ASD have a different reading profile from other reading disorders that needs to be specifically targeted in interventions.


Natural and anthropogenic factors shape metazoan parasite community structure in mummichog (Fundulus heteroclitus) from two estuaries in New Brunswick, Canada.
Previously published multidisciplinary studies in the Miramichi and Bouctouche rivers (New Brunswick, Canada) noted significant changes in fish health parameters, including elevated tissue levels of organic contaminants and a wide range of physiological disturbances, in mummichog Fundulus heteroclitus (L.) from a site on the Miramichi River that received bleached kraft pulpmill and municipal effluent. The present study reports differences in the abundance of individual parasite species, as well as parasite infracommunity and component community composition, in mummichog from both rivers. These differences were evaluated in relation to host (size, condition, immune function, tissue organochlorine contaminant levels) and environmental (faecal coliform counts, salinity, temperature) data derived from the previously published studies. Overall, 18 parasite species were identified, the most common of which were Ascocotyle sp. larv., Ornithodiplostomum sp. larv., Posthodiplostomum sp. larv., and Proteocephalus filicollis (Rudolphi, 1802). There were broad differences in parasite community structure and composition between rivers and within rivers, the most prominent pattern being a pronounced difference between sites in the upper and lower estuary of each river that was likely driven by salinity. Mean infracommunity richness was also positively related to faecal coliforms (considered here as a surrogate measure of eutrophication via municipal sewage), and both were highest at the most polluted site. We noted no other significant relationships. Thus our data suggest that the parasite communities in these two estuaries were primarily structured by large upstream/downstream ecological gradients in salinity, and secondarily by eutrophication due to pollution by municipal and industrial effluents. Overall, our results highlight the value of coordinated multidisciplinary studies for understanding the factors that shape parasite abundance and community structure.


Superoxide dismutase from the eukaryotic thermophile Alvinella pompejana: structures, stability, mechanism, and insights into amyotrophic lateral sclerosis.
Prokaryotic thermophiles supply stable human protein homologs for structural biology; yet, eukaryotic thermophiles would provide more similar macromolecules plus those missing in microbes. Alvinella pompejana is a deep-sea hydrothermal-vent worm that has been found in temperatures averaging as high as 68 degrees C, with spikes up to 84 degrees C. Here, we used Cu,Zn superoxide dismutase (SOD) to test if this eukaryotic thermophile can provide insights into macromolecular mechanisms and stability by supplying better stable mammalian homologs for structural biology and other biophysical characterizations than those from prokaryotic thermophiles. Identification, cloning, characterization, X-ray scattering (small-angle X-ray scattering, SAXS), and crystal structure determinations show that A. pompejana SOD (ApSOD) is superstable, homologous, and informative. SAXS solution analyses identify the human-like ApSOD dimer. The crystal structure shows the active site at 0.99 A resolution plus anchoring interaction motifs in loops and termini accounting for enhanced stability of ApSOD versus human SOD. Such stabilizing features may reduce movements that promote inappropriate intermolecular interactions, such as amyloid-like filaments found in SOD mutants causing the neurodegenerative disease familial amyotrophic lateral sclerosis or Lou Gehrig's disease. ApSOD further provides the structure of a long-sought SOD product complex at 1.35 A resolution, suggesting a unified inner-sphere mechanism for catalysis involving metal ion movement. Notably, this proposed mechanism resolves apparent paradoxes regarding electron transfer. These results extend knowledge of SOD stability and catalysis and suggest that the eukaryote A. pompejana provides macromolecules highly similar to those from humans, but with enhanced stability more suitable for scientific and medical applications.


Brief educational "curbside consultation": Impact on attention-deficit/hyperactivity disorder referrals in an integrated healthcare setting.
As behavioral health providers integrate into primary health care clinics, it is important to examine methods used to increase primary care providers' (PCPs) knowledge of behavioral health providers' areas of expertise and service provision. Attention-deficit/hyperactivity disorder (ADHD) has been identified as one of the most common behavioral health disorders PCPs diagnose and treat in children. This study examined whether 2 brief educational "curbside consults," during which a psychologist provided information regarding the assessment and treatment of ADHD, had an impact on PCPs' referral practices. Information was collected via medical records review for the same 6-month period prior to and following provider education. The results indicated that there was an increase in ADHD referrals to the behavioral health clinic (BHC) following educational consultation. In addition, there was some evidence to suggest that following PCP education, fewer children were diagnosed by the PCP with ADHD and fewer children were previously prescribed a psychotropic medication upon referral to the BHC. Treatment (e.g., behavioral therapy, pharmacotherapy, combined approach) also differed between families previously diagnosed and prescribed psychotropic medication. The current study provides some preliminary evidence that brief educational consultation with PCPs may increase both referrals for ADHD concerns and may influence the way PCPs diagnose and address behavioral health concerns. Implications of these findings and suggestions for future research are discussed. (PsycINFO Database Record


Prevalence of psychomorbidity among patients with chronic cough.
Chronic cough may cause significant emotional distress and although patients are not routinely assessed for co-existent psychomorbidity, a cough that is refractory to any treatment is sometimes suspected to be functional in origin. It is not known if patients with chronic cough referred for specialist evaluation have emotional impairment but failure to recognise this may influence treatment outcomes. In this cross-sectional study, levels of psychomorbidity were measured in patients referred to a specialist cough clinic. Fifty-seven patients (40 female), mean age 47.5 (14.3) years referred for specialist evaluation of chronic cough (mean cough duration 69.2 (78.5) months) completed the Hospital Anxiety and Depression (HAD) scale, State Trait Anxiety Inventory (STAI) and the Crown Crisp Experiential Index (CCEI) at initial clinic presentation. Subjects then underwent a comprehensive diagnostic evaluation, after which they were classified as either treated cough (TC) or idiopathic cough (IC). Questionnaire scores were compared between TC (n = 42) and IC (n = 15). Using the HAD scale, 33% of all cough patients were identified as anxious, while 16% experienced depression. The STAI scores suggested moderate or high trait anxiety in 48% of all coughers. Trait anxiety was significantly higher among TC (p < 0.001) and IC patients (p = 0.004) compared to a healthy adult population. On the CCEI, mean scores on the phobic anxiety, somatisation, depression, and obsession subscales were significantly higher among all cough patients than the published mean scores for healthy controls. Only state anxiety was significantly higher in IC patients compared with TC patients (p < 0.05). Patients with chronic cough appear to have increased levels of emotional upset although psychological questionnaires do not readily distinguish between idiopathic coughers and those successfully treated.


Glycyrrhizin inhibits porcine epidemic diarrhea virus infection and attenuates the proinflammatory responses by inhibition of high mobility group box-1 protein.
Porcine epidemic diarrhea (PED), caused by porcine epidemic diarrhea virus (PEDV) infection, leads to significant economic losses in the swine industry worldwide. In our studies, we found that glycyrrhizin, the major component of licorice root extracts, could moderately inhibit PEDV infection in Vero cells, when analyzed by western blot, qRT-PCR and a plaque formation assay. We also revealed that glycyrrhizin inhibited the entry and replication of PEDV. In addition, we demonstrated that glycyrrhizin decreased the mRNA levels of proinflammatory cytokines. Since glycyrrhizin is a competitive inhibitor of high mobility group box-1 (HMGB1), we confirmed that TLR4 and RAGE (£ associated with PEDV pathogenesis during the infection in Vero cells. In summary, our studies provide a molecular basis for developing novel therapeutic methods to control PEDV infection, based on glycyrrhizin and its derivatives.


Bacteriological Profile of Diabetic Foot Ulcer with Special Reference to Drug-resistant Strains in a Tertiary Care Center in North-East India.
This study was carried out to determine the bacteriological profile of infected diabetic foot ulcers (DFUs) and the antibiotic resistance pattern from the isolates. An attempt was made to suggest an empiric antibiotic regimen to treat such patients. Tissue samples were collected from 150 patients between February 2015 and January 2016 with DFUs under aseptic precautions and they were processed as per the Clinical and Laboratory Standards Institute guidelines. A total of 185 bacterial isolates were obtained from 150 persons with diabetic and with foot ulcers. The age group of these persons ranged from 35 to 80 years and the maximum number of persons with DFUs was in the age group of 60-65 years. Among the isolates, Gram-negative bacilli were isolated in 112/185 (61%) and Gram-positive cocci in 73/185 (39%) cases. The most common isolate was Staphylococcus spp. 46 (25%), followed by Escherichia coli (20%) and Enterococcus spp. (15%). The antibiotic sensitivity profile of the bacteria was also studied. Among the isolates, 59/112 (53%) of the Gram-negative bacilli were extended spectrum beta-lactamase producers, 19/46 (41%) were methicillin-resistant Staphylococcus aureus, and 5/27 (19%) were vancomycin-resistant Enterococcus. This study showed a preponderance of multidrug-resistant strains among the isolates from the DFUs. Knowledge on the antibiotic sensitivity pattern of the isolates will be helpful in determining the drugs for the empirical treatment of diabetic ulcers. Thus, indiscriminate use of antibiotics and chances of subsequent development of antibiotic resistance can also be reduced.


ETIOPATHOGENIC, EPIDEMIOLOGIC AND CLINICAL-THERAPEUTIC COMPARISON OF NON-HODGKIN'S LYMPHOMA AND KAPOSI'S SARCOMA.
Non-Hodgkin's lymphomas (NHL) are primary neoplasms derived from lymphocytes, and Kaposi's sarcoma (SK) is a multicentric disease of viral etiology and is associated with HIV. To study the etiopathogenesis and clinical characteristics of NHL and KS, describing their mutual factors. This retrospective investigation was performed on 101 medical charts. The patients were studied according to their age, gender, and HIV-positivity, following the PRISMA guidelines. The characteristics of the tumors and comorbidities were analyzed according to their age and lymphatic metastasis. The mean age of the patients ranged between 15-87 years for NHL and between 25-54 for KS, but the age of patients with NHL associated with HIV did not surpass 34 years. The ratio male: female was 1,8:1 for NHL, but only men presented KS. HIV-positivity was found in five patients with NHL and in 14 with KS. The stages of NHL were: I (21%), II (18,4%), III (26,3%), and IV (34,2%), but KS were found only at III (40%) and IV (60%) stages. The lymphatic metastases were positive in 62 patients NHL and in four with KS. HIV-positivity occurred in 60% of patients with NHL and in 50% with KS. The HIV seropositivity was revealed for most of patients during the NHL and SK propaedeutic and none of them present clinical manifestations of AIDS. NHL associated with HIV was found only in young patients. NHL and KS patients have similar epidemiological, clinical, and therapeutic characteristics.


Diffusible signal factor-mediated quorum sensing plays a central role in coordinating gene expression of Xanthomonas citri subsp. citri.
Diffusible signal factor (DSF) family signal-mediated quorum sensing (QS) has been identified in many gram-negative bacteria. This QS pathway of Xanthomonas spp. consists of three major QS components: RpfF, RpfC, and RpfG. The rpfF gene encodes a putative enoyl-CoA hydratase that catalyzes the synthesis of the signal molecule. RpfC and RpfG serve as a two-component system for the perception and transduction of the extracellular DSF family signals. In order to further characterize the QS regulatory network in Xanthomonas citri subsp. citri, we investigated the RpfF, RpfC, and RpfG regulons by using transcriptome analyses. Comparison of the transcriptomes of the QS mutants (rpfF, rpfC, and rpfG) with that of the wild-type strain revealed a core group of genes controlled by all three QS components, suggesting that the RpfC-RpfG two-component system is a major and conserved signal perception and transduction system for DSF family signal-mediated QS in X. citri subsp. citri. The unique genes controlled by RpfF alone indicate the complexity of the QS pathway and the involvement of additional sensory mechanisms in X. citri subsp. citri. The unique genes controlled by RpfC and RpfG, respectively, support the possibility that RpfC and RpfG play broader roles in gene regulation other than transduction of DSF signals.


Older adults at high risk of HIV infection in China: a systematic review and meta-analysis of observational studies.
There is an increasing prevalence of human immunodeficiency virus (HIV) infection in older adults in China, but the findings across prevalence studies have been mixed. This is the first meta-analysis of the prevalence of HIV infection and its moderating factors in older adults in China. Two investigators systematically and independently searched both international (PubMed, PsycINFO, Web of Sciences and EMBASE) and Chinese (WanFang, CNKI, and CQVIP) databases. HIV infection rates in older adults were analyzed using the random-effects model. Altogether 46 studies were included in the analysis. The pooled prevalence of HIV infection in older adults was 2.1% (95% CI [1.9%-2.3%], I 2 = 99.4%). Subgroup analyses revealed that men who have sex with men (MSM), hospital population samples, publications after 2014, studies conducted in the western region of China, and higher study quality were significantly associated with higher HIV infection rate. This meta-analysis found that the HIV infection prevalence in older adults is significantly higher than the general population in China. Attention should be given to this urgent public health issue, and effective HIV/AIDS preventive, screening and treatment measures are warranted in this population. PROSPERO: CRD42019124286.


Serine esterase and hemolytic activity in human cloned cytotoxic T lymphocytes.
Target cell lysis by most murine cytotoxic T lymphocytes appears to be mediated by a complement (C9)-like protein called perforin, contained in high-density cytoplasmic granules. These granules also contain high levels of serine esterase activity, which may also play a role in cytolysis. Analysis of 17 cloned human cytotoxic T lymphocytes revealed the presence of serine esterase that is very similar to its murine counterpart in substrate and inhibitor specificities, pH optimum, and molecular mass; dot blot hybridization with synthetic oligonucleotides corresponding to the active sites of two known murine CTL esterases suggests homology to the murine enzyme HF. However, serine esterase was present at only approximately 10% of the level found in murine CTLs, and was not secreted during CTL-target cell interaction; moreover, hemolytic activity could not be detected in any of the seven cell lines tested. The results suggest that the human CTLs examined here kill their target cells by a mechanism different from that used by most cloned murine CTLs.


Improved therapy for medulloblastoma: targeting hedgehog and PI3K-mTOR signaling pathways in combination with chemotherapy.
Aberrant activation and interactions of hedgehog (HH) and PI3K/AKT/mTOR signaling pathways are frequently associated with high-risk medulloblastoma (MB). Thus, combined targeting of the HH and PI3K/AKT/mTOR pathways could be a viable therapeutic strategy to treat high-risk patients. Therefore, we investigated the anti-MB efficacies of combined HH inhibitor Vismodegib and PI3K-mTOR dual-inhibitor BEZ235 together or combined individually with cisplatin against high-risk MB. Using non-MYC- and MYC-amplified cell lines, and a xenograft mouse model, the in vitro and in vivo efficacies of these therapies on cell growth/survival and associated molecular mechanism(s) were investigated. Results showed that combined treatment of Vismodegib and BEZ235 together, or with cisplatin, significantly decreased MB cell growth/survival in a dose-dependent-fashion. Corresponding changes in the expression of targeted molecules following therapy were observed. Results demonstrated that inhibitors not only suppressed MB cell growth/survival when combined, but also significantly enhanced cisplatin-mediated cytotoxicity. Of these combinations, BEZ235 exhibited a significantly greater efficacy in enhancing cisplatin-mediated MB cytotoxicity. Results also demonstrated that the MYC-amplified MB lines showed a higher sensitivity to combined therapies compared to non-MYC-amplified cell lines. Therefore, we tested the efficacy of combined approaches against MYC-amplified MB growing in NSG mice. In vivo results showed that combination of Vismodegib and BEZ235 or their combination with cisplatin, significantly delayed MB tumor growth and increased survival of xenografted mice by targeting HH and mTOR pathways. Thus, our studies lay a foundation for translating these combined therapeutic strategies to the clinical setting to determine their efficacies in high-risk MB patients.


Specialized postsynaptic morphology enhances neurotransmitter dilution and high-frequency signaling at an auditory synapse.
Sensory processing in the auditory system requires that synapses, neurons, and circuits encode information with particularly high temporal and spectral precision. In the amphibian papillia, sound frequencies up to 1 kHz are encoded along a tonotopic array of hair cells and transmitted to afferent fibers via fast, repetitive synaptic transmission, thereby promoting phase locking between the presynaptic and postsynaptic cells. Here, we have combined serial section electron microscopy, paired electrophysiological recordings, and Monte Carlo diffusion simulations to examine novel mechanisms that facilitate fast synaptic transmission in the inner ear of frogs (Rana catesbeiana and Rana pipiens). Three-dimensional anatomical reconstructions reveal specialized spine-like contacts between individual afferent fibers and hair cells that are surrounded by large, open regions of extracellular space. Morphologically realistic diffusion simulations suggest that these local enlargements in extracellular space speed transmitter clearance and reduce spillover between neighboring synapses, thereby minimizing postsynaptic receptor desensitization and improving sensitivity during prolonged signal transmission. Additionally, evoked EPSCs in afferent fibers are unaffected by glutamate transporter blockade, suggesting that transmitter diffusion and dilution, and not uptake, play a primary role in speeding neurotransmission and ensuring fidelity at these synapses.


A membrane-associated progesterone-binding protein, 25-Dx, is regulated by progesterone in brain regions involved in female reproductive behaviors.
The ventromedial hypothalamus (VMH) plays a central role in the regulation of the female reproductive behavior lordosis, a behavior dependent upon the sequential activation of receptors for the ovarian steroid hormones estradiol (E) and progesterone (P). These receptors function as transcription factors to alter the expression of target genes. To discover behaviorally relevant genes targeted by E and P in the VMH, we used the differential display PCR to identify messenger RNAs that are differentially expressed in the hypothalamus of ovariectomized (ovx) rats treated with E alone compared with ovariectomized rats treated with E and P. We show here that one interesting mRNA within the hypothalamus that is repressed by P after E priming encodes the protein 25-Dx, the rat homolog of the human membrane-associated P-binding protein Hpr6.6. Neurons in the brain containing the highest levels of 25-Dx are located in several nuclei of the basal forebrain, including the VMH. 25-Dx expression is also higher in the hypothalamus of female P receptor "knockout" mice than in their wild-type littermates. These findings suggest a mechanism in which the activation of nuclear P receptor represses expression of a membrane P receptor, 25-Dx, during lordosis facilitation.


Electromyographic evaluation of muscular work pattern as a predictor of trapezius myalgia.
Electromyographic (EMG) measurements and interviews concerning muscular pain and disability were performed prospectively every 10th week on 30 healthy new female employees of a chocolate manufacturing plant. The static muscle activity and rate of short unconscious interruptions in EMG activity (EMG gaps) of the trapezius muscle during repetitive work tasks were evaluated as possible risk factors for patient status with trapezius myalgia. At the start of employment, the static and median contraction levels were significantly higher in future patients than in the rest of the subjects (nonpatients). In subsequent recordings these values were reduced to the level of the nonpatients. Throughout the study, the future patients had a lower frequency of EMG gaps than the nonpatients. A regression analysis showed a significant value for a low rate of EMG gaps to predict future patient status.


Thymus vulgaris L. and thymol assist murine macrophages (RAW 264.7) in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans.
Microorganisms are capable to combat defense cells by means of strategies that contribute to their stabilization and proliferation in invaded tissues. Frequently antimicrobial-resistant strains appear; therefore, alternative methods to control them must be investigated, for example, the use of plant products. The capacity of the thyme extract (Thymus vulgaris L.) and phytocompound thymol in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in murine macrophages (RAW 264.7) was evaluated. Minimal inhibitory concentrations (MIC) of the plant products were used. The effect of these MIC were analyzed in the assays of phagocytosis and immunoregulation by analysis of the production of cytokines (IL-1β, TNF-α, and IL-10) and nitric oxide (NO). The plant products effectively assisted the macrophages in the phagocytosis of microorganisms, presenting significant reductions of S. aureus and P. aeruginosa. The macrophages also regulated the production of inflammatory mediators in the infections by S. aureus, P. aeruginosa, and C. albicans. In addition, thyme provided a satisfactory effect in response to the bacterial infections, regarding generation of NO. Thus, the effectiveness of the thyme and thymol to control in vitro infections by S. aureus, P. aeruginosa, and C. albicans was observed. Phagocytosis of S. aureus by RAW 264.7 was enhanced with thymol Thyme enhanced the phagocytosis of P. aeruginosa by RAW 264.7 Plant products provided immunoregulation of inflammatory cytokines Production of nitric oxide was improved with the treatments in bacterial infections.


Conformational analysis of endothelin-1 analogs with indolizidinone amino acids incorporated at the C-terminus.
In light of the fact that a beta-turn conformation at the C-terminus of endothelin-1 (ET-1) could be responsible for activity at the ET(B) receptor, the incorporation of (3S, 6S, 9S)-2-oxo-3-amino-1-azabicyclo[4.3.0]nonane-9-carboxylic acid (IAA) should provide an elegant method to establish whether a formyl group on Trp21 plays a role in stabilizing a beta-turn. Eight linear ET-1 analogs, four formylated and four nonformylated, ET-1-(Leu17-Asp18-IAA-Trp21); ET-1-(Leu17-IAA-Ile20-Trp21); ET-1-(Leu17-Asp18-Pro19-Ile20-Trp21) and ET-1-(Leu17-Asp 8-Ile19-Pro20-Trp21) have been analyzed by high-resolution nuclear magnetic resonance (NMR) spectroscopy and molecular modeling. Two-dimensional double quantum filtered correlation spectroscopy (DQFCOSY), total correlation spectroscopy (TOCSY) and nuclear Overhausen enhancement spectroscopy (NOESY) were resolved and analyzed for each molecule. Interspatial distance constraints were derived from the intensity of the NOESY connectivities, The formation of hydrogen bonding was monitored from the temperature-dependence of the NH chemical shifts. Molecular models calculated by means of distance geometry, simulated annealing and energy minimization, suggested a global elongated structure for the formylated analogs and a folded arrangement for the nonformylated derivatives, but no hydrogen bonding was detected at the C-terminus of ET-1 analogs.


Time-reversal symmetry and universal conductance fluctuations in a driven two-level system.
In the presence of time-reversal symmetry, quantum interference gives strong corrections to the electric conductivity of disordered systems. The self-interference of an electron wave function traveling time-reversed paths leads to effects such as weak localization and universal conductance fluctuations. Here, we investigate the effects of broken time-reversal symmetry in a driven artificial two-level system. Using a superconducting flux qubit, we implement scattering events as multiple Landau-Zener transitions by driving the qubit periodically back and forth through an avoided crossing. Interference between different qubit trajectories gives rise to a speckle pattern in the qubit transition rate, similar to the interference patterns created when coherent light is scattered off a disordered potential. Since the scattering events are imposed by the driving protocol, we can control the time-reversal symmetry of the system by making the drive waveform symmetric or asymmetric in time. We find that the fluctuations of the transition rate exhibit a sharp peak when the drive is time symmetric, similar to universal conductance fluctuations in electronic transport through mesoscopic systems.


Towards a risk map of malaria for Sri Lanka: the importance of house location relative to vector breeding sites.
In Sri Lanka, the major malaria vector Anopheles culicifacies breeds in pools formed in streams and river beds and it is likely that people living close to such breeding sites are at higher risk of malaria than people living further away. This study was done to quantify the importance of house location relative to vector breeding sites for the occurrence of malaria in order to assess the usefulness of this parameter in future malaria risk maps. Such risk maps could be important tools for planning efficient malaria control measures. In a group of seven villages in north central Sri Lanka, malaria cases were compared with community controls for distance from house to breeding sites and a number of other variables, including type of housing construction and use of anti-mosquito measures. The presence of An. culicifacies in bedrooms was determined by indoor insecticide spray collections. People living within 750 m of the local stream, which was the established vector-breeding site, were at much higher risk for malaria than people living further away (odds ratio adjusted for confounding by other variables 5.93, 95% CI: 3.50-8.91). Houses close to the stream also had more adult An. culicifacies in the bedrooms. Poor housing construction was an independent risk factor for malaria. Risk maps of malaria in Sri Lanka can be based on the location of houses relative to streams and rivers that are potential breeding sites for the malaria vector An. culicifacies. A distance of 750 m is suggested as the cut-off point in defining low- and high-risk villages.


Properties of single potassium channels modulated by glucose in rat pancreatic beta-cells.
1. The patch clamp method has been used to examine the effect of glucose on single K+ channel currents recorded from cell-attached patches on dissociated rat pancreatic beta-cells. Patch pipettes contained a 140 mM-K+ solution. 2. In glucose-free solution three types of K+ channels were observed. Two of these, having conductances of around 50 pS (G-channel) and 20 pS when the external K+ concentration, [K+]0, was 140 mM, were active at the resting potential of the cell. The G-channel was observed in more patches and showed higher activity; it therefore appears to contribute the major fraction of the resting K+ permeability of the beta-cell. At membrane potentials positive to about +20 mV a third type of K+ channel, having a mean conductance of 120 pS, was activated. The open probability of this channel was strongly voltage dependent and increased with depolarization. 3. The reversal potential of the G-channel current was shifted 59 mV by a 10-fold change in external K+ (Na+ substitution) indicating the channel is highly K+ selective. The single-channel conductance varied with [K+]o as predicted from the Goldman-Hodgkin-Katz equation; at physiological [K+]o (5 mM-K+) an inward conductance of around 10 pS is predicted. The amplitude of the single-channel current showed a tendency to saturate with increasing [K+]o. 4. Single G-channel currents show burst kinetics indicating at least two closed states. The open and closed (gap) times within the bursts were distributed exponentially with time constants of 2.5 ms (tau o) and 0.5 ms (tau c1) respectively at the resting potential of the cell. There was little change in tau c1 over the voltage range -40 to 60 mV (pipette potential) but tau o increased slightly with membrane depolarization. 5. The addition of glucose to the bath solution produced a reversible, dose-dependent decrease in G-channel activity. This decrease results principally from a reduction in the frequency and duration of the bursts of openings with increasing glucose. In addition, the mean open time decreases. The short gaps during the bursts were little affected by glucose. 6. At glucose concentrations of .10 mM and above the decrease in G-channel activity is accompanied by an increase in the input resistance of the cell and by the initiation of action potentials. 7. It is concluded that glucose metabolism results in a reduction of G-channel open probability and thereby produces depolarization of the beta-cell.


Tertiary lymphoid organs in the inflammatory myopathy associated with PD-1 inhibitors.
Programmed cell death 1 inhibitors have revolutionized therapy for cancer by their outstanding effectiveness. However, they may cause adverse effects, among which inflammatory myopathy is one of the most disabling. To elucidate its mechanism, we analysed muscle biopsies and compared them with other inflammatory myopathies. Muscle biopsies from three patients with inflammatory myopathy after treatment with PD-1 inhibitors for cancer were subjected to immunohistochemical and ultrastructural analyses to localize CD8+ cytotoxic cells and markers of lymphoid follicles. For comparison, two cases of polymyositis and one of juvenile dermatomyositis were examined. Nearly identical pathological features were observed in the three cases. In the island-like foci of inflammation, muscle fibers were undergoing degeneration. CD8+ cytotoxic T cells, macrophages, CD4+ cells, and B cells were observed in the foci. CD8+ cells were seen outside and inside the basal lamina of non-necrotic muscle fibers. Lymphoid follicle-like structures with CD21+ follicular dendritic cells were present. The blood vessels in the foci showed features consistent with the high endothelial venules, on which their markers, PNAd and CCL21, were expressed. In polymyositis, blood vessels stained only faintly for PNAd and CCL21, while in juvenile dermatomyositis, in which tertiary lymphoid follicle-like structure was reported in the past, they stained positively. In inflammatory myopathy associated with PD-1 inhibitors, CD8+ cells appear to predominantly destruct muscle fibers. The presence of lymphoid follicle-like structures and expression of PNAd and CCL21 on the endothelial cells suggest the tertiary lymphoid organs are formed, and involved in the leakage of lymphocytes. Thus, in the three cases examined, formation of the tertiary lymphoid organs is likely to play an important role in genesis of the PD-1 myopathy.


Correction of altered noradrenaline reactivity in essential hypertension by indapamide.
Fourteen patients with untreated mild to moderate essential hypertension had, on average, an abnormally high cardiovascular reactivity to exogenous noradrenaline and angiotensin II, while plasma noradrenaline, renin activity, exchangeable body sodium, and blood volume were normal. Treatment with a low dose of indapamide (2.5 mg/day) for 6 weeks decreased blood pressure by 10% in these hypertensive patients but not in 13 normal control subjects. Plasma or blood volume and exchangeable sodium were not changed significantly; nevertheless, the latter, and body weight, tended to be decreased slightly. Though a mild reduction in extracellular sodium in both normal and hypertensive subjects appears possible, it may not fully explain per se the blood pressure-lowering effect of indapamide in essential hypertension. Indapamide induced a mild decrease in angiotensin II pressor responsiveness in normal or hypertensive subjects, but a possible depressor influence from this change was probably antagonized by a concomitant pronounced increase in plasma renin activity. In hypertensive patients, the abnormally high noradrenaline reactivity was corrected by indapamide without an accompanying increase in endogenous plasma noradrenaline levels. Indapamide-induced changes in blood pressure correlated with those in noradrenaline pressor dose. It was concluded, therefore, that indapamide may decrease blood pressure in essential hypertension at least in part by lowering an abnormally high cardiovascular noradrenaline reactivity without causing an equivalent increase in adrenergic nervous activity.


Parallel preparation of densely packed arrays of 150-nm gold-nanocrescent resonators in three dimensions.
Metallic nanostructures show interesting optical properties due to their plasmonic resonances, and when arranged in three-dimensional (3D) arrays hold promise for optical metamaterials with negative refractive index. Towards this goal a simple, cheap, and parallel method to fabricate large-area, ordered arrays of 150-nm gold nanocrescents supporting plasmonic resonances in the near-infrared spectral range is demonstrated. In this process hexagonally ordered monolayers of monodisperse colloids are prepared by a simple floating technique, and subsequently the individual particles are size-reduced in a plasma process and used as a shadow mask with the initial lattice spacing. The resulting two-dimensional array of plasmonic resonators is coated with a transparent silica layer, which serves as a support for a second layer prepared by the identical process. The mutual orientation of the nanostructures between the individual layers can be freely adjusted, which determines the polarization-dependent absorption of the array and opens the possibility to introduce chirality in this type of 3D metamaterial. The iteration of this simple and efficient methodology yields 3D arrays with optical features as sharp as those of the individual nanocrescents, and shows strong potential for large-scale production of high-quality optical metamaterials.


Do changes in surgical procedures for breast cancer have consequences for hospital mean length of stay? A study of women operated on for breast cancer in Sweden, 1980-95.
Between 1986 and 1996, the overall mean overnight length of stay for all diagnoses in Sweden decreased from 20.8 to 7.1 days. The study describes changes in surgical technique, from mastectomy to breast-conserving surgery, in treatment of female breast cancer and the parallel change in average length of hospital stay, and discusses the possible link between the trends. The study was performed as a descriptive register study on hospital admission data from the Swedish Hospital Discharge Register over a 16-year period (1980-95). During the study period, the mean length of stay for surgical curative breast cancer treatment in Sweden decreased by 56%. In 1980, the proportion of women receiving conservative surgery was 7%. At the end of the period, this share had increased to 51%. Breast-conserving surgery had an approximately 30% shorter mean length of stay compared with mastectomy. The gap was remarkably stable during the study period. The shift from mastectomy to breast-conserving surgery had a limited effect on the share of patients that went through lymph node dissection. Neither age nor the number of operations per woman could, to any significant extent, explain the decrease in mean length of stay. Approximately 14% of the overall decline can be attributed to the changes in technique. Clinical practice style, in this case the surgical technique, has had an effect on length of stay, but the surgical technique can only to some extent explain the trend.


Setting a standard of affordability for health insurance coverage.
Recently, Massachusetts passed landmark legislation designed to expand health insurance coverage. This legislation includes a requirement that all adults enroll in a health insurance plan. This mandate takes effect only if an "affordable" plan is available. The definition of affordability for individuals and families of different incomes or circumstances is a critical decision in implementation and is relevant to any state or federal reform requiring individual premium or cost-sharing contributions, or both. This analysis was done to assist the policy design process in Massachusetts and delineates an empirically based approach to setting affordability standards.


Conformational transitions of membrane-bound HIV-1 fusion peptide.
The human immunodeficiency virus type-1 (HIV-1) fusion peptide (FP) functions as a non-constitutive membrane anchor that translocates into membranes during envelope glycoprotein-induced fusion. Here, by means of infrared spectroscopy (IR) and of various bilayer-perturbation assays, we describe the peptide conformations that are accessible to its membrane-bound state and the transitions occurring between them. The peptide underwent a conformational transition from a predominantly alpha-helical structure to extended beta-type strands by increasing peptide concentration in 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG) vesicles. A comparable transition was observed at a fixed 1:100 peptide-to-lipid ratio when calcium was added to vesicles containing prebound alpha-helical peptide. Cation binding induced an increase in the amount of H-bonded carbonyls within the interfacial region of POPG. Calcium-promoted alpha-->beta conversion in membranes correlated with the closure of preformed lytic pores and took place in dispersed (nonaggregated) vesicles doped with poly(ethylene glycol)-lipid conjugates, showing that the conformational transition was independent of vesicle aggregation. We conclude that the target membrane conditions modulate the eventual structure adopted by the HIV-1 FP. Conformational polymorphism of the inserted peptide may contribute to the flexibility of the fusogenic complex during the fusion reaction cycle, and/or may be related to target membrane perturbation at the fusion locus.