Special education for students with disabilities: analysis and recommendations.
Twenty years ago, the educational rights of students with disabilities were dramatically and firmly established in law and practice. Prior to that time, many students were refused enrollment or special educational services. As recently as 1973, at least one million students were denied enrollment in public schools solely on the basis of their disabilities, and at least two million others were not receiving an education appropriate to their needs. Although every state has provided some form of special education throughout this century, these services were largely at the discretion of local school districts. Only since a federal court case in 1972 and the passage of federal legislation in 1975 have all states been mandated to provide a free, appropriate public education to all students with disabilities. Today, as Parrish and Chambers point out in this journal issue, special education for students with disabilities is the largest categorical program in public schools, costing an estimated $32 billion. Since the passage of Public Law 94-142 in 1975 (later retitled the Individuals with Disabilities Education Act, or the IDEA), the number of elementary and secondary students receiving special education has increased from 3.7 million to 4.6 million, increasing also from 8% to 11% of all students in public schools. According to Parrish and Chambers, the population of students eligible for special education is expected to continue to rise. The IDEA governs the educational rights of individuals from birth to age 21, though only students in elementary and secondary school are addressed in this journal issue. The IDEA allowed access to the public schools for many students who had previously been denied enrollment. The IDEA has also been given partial credit for decreasing the rate of institutionalization of individuals with disabilities. Before the IDEA, many parents had the sole responsibility of meeting all the needs of their severely disabled children 24 hours per day; once schools began to provide extensive services to students with severe disabilities, more families were able to avoid institutionalization. Under the IDEA, states and local districts were given a mandate to provide specialized educational programs to students with special needs, and students and parents were given a mechanism for enforcement of their rights. In a 1989 survey, 94% of parents of students with disabilities agreed that services for these students had improved since the implementation of the IDEA. Yet special education today is widely criticized as expensive, ineffective, inadequately coordinated with regular education, and/or culturally biased. The National Association of State Boards of Education has recommended radical reduction in the size of special education. Special education also has its champions, who argue that many students perform better academically and have better self-esteem when provided with special services, often in a separate setting. This analysis addresses five questions concerning special education under the IDEA: (1) Why are so many students considered disabled? (2) What are the educational needs of students with disabilities? (3) How should appropriate, individualized services be funded? (4) Are the IDEA's procedural protections necessary? (5) Can regular education meet the needs of more students?


From bench to bedside: The mGluR5 system in people with and without Autism Spectrum Disorder and animal model systems.
The metabotropic glutamate receptor 5 (mGluR5) is a key regulator of excitatory (E) glutamate and inhibitory (I) γ-amino butyric acid (GABA) signalling in the brain. Despite the close functional ties between mGluR5 and E/I signalling, no-one has directly examined the relationship between mGluR5 and glutamate or GABA in vivo in the human brain of autistic individuals. We measured [18F] FPEB (18F-3-fluoro-5-[(pyridin-3-yl)ethynyl]benzonitrile) binding in 15 adults (6 with Autism Spectrum Disorder) using two regions of interest, the left dorsomedial prefrontal cortex and a region primarily composed of left striatum and thalamus. These two regions were mapped out using MEGA-PRESS voxels and then superimposed on reconstructed PET images. This allowed for direct comparison between mGluR5, GABA + and Glx. To better understand the molecular underpinnings of our results we used an autoradiography study of mGluR5 in three mouse models associated with ASD: Cntnap2 knockout, Shank3 knockout, and 16p11.2 deletion. Autistic individuals had significantly higher [18F] FPEB binding (t (13) = -2.86, p = 0.047) in the left striatum/thalamus region of interest as compared to controls. Within this region, there was a strong negative correlation between GABA + and mGluR5 density across the entire cohort (Pearson's correlation: r (14) = -0.763, p = 0.002). Cntnap2 KO mice had significantly higher mGlu5 receptor binding in the striatum (caudate-putamen) as compared to wild-type (WT) mice (n = 15, p = 0.03). There were no differences in mGluR5 binding for mice with the Shank3 knockout or 16p11.2 deletion. Given that Cntnap2 is associated with a specific striatal deficit of parvalbumin positive GABA interneurons and 'autistic' features, our findings suggest that an increase in mGluR5 in ASD may relate to GABAergic interneuron abnormalities.


Antigen receptor-induced activation and cytoskeletal rearrangement are impaired in Wiskott-Aldrich syndrome protein-deficient lymphocytes.
The Wiskott-Aldrich syndrome protein (WASp) has been implicated in modulation of lymphocyte activation and cytoskeletal reorganization. To address the mechanisms whereby WASp subserves such functions, we have examined WASp roles in lymphocyte development and activation using mice carrying a WAS null allele (WAS(-)(/)(-)). Enumeration of hemopoietic cells in these animals revealed total numbers of thymocytes, peripheral B and T lymphocytes, and platelets to be significantly diminished relative to wild-type mice. In the thymus, this abnormality was associated with impaired progression from the CD44(-)CD25(+) to the CD44(-)CD25(-) stage of differentiation. WASp-deficient thymocytes and T cells also exhibited impaired proliferation and interleukin (IL)-2 production in response to T cell antigen receptor (TCR) stimulation, but proliferated normally in response to phorbol ester/ionomycin. This defect in TCR signaling was associated with a reduction in TCR-evoked upregulation of the early activation marker CD69 and in TCR-triggered apoptosis. While induction of TCR-zeta, ZAP70, and total protein tyrosine phosphorylation as well as mitogen-activated protein kinase (MAPK) and stress-activated protein/c-Jun NH(2)-terminal kinase (SAPK/JNK) activation appeared normal in TCR-stimulated WAS(-)(/)(-) cells, TCR-evoked increases in intracellular calcium concentration were decreased in WASp-deficient relative to wild-type cells. WAS(-)(/)(-) lymphocytes also manifested a marked reduction in actin polymerization and both antigen receptor capping and endocytosis after TCR stimulation, whereas WAS(-)(/)(-) neutrophils exhibited reduced phagocytic activity. Together, these results provide evidence of roles for WASp in driving lymphocyte development, as well as in the translation of antigen receptor stimulation to proliferative or apoptotic responses, cytokine production, and cytoskeletal rearrangement. The data also reveal a role for WASp in modulating endocytosis and phagocytosis and, accordingly, suggest that the immune deficit conferred by WASp deficiency reflects the disruption of a broad range of cellular behaviors.


A Nested PCR Assay for Detecting Valsa mali var. mali in Different Tissues of Apple Trees.
A nested polymerase chain reaction (PCR) assay for detecting Valsa mali var. mali, the causal agent of apple tree Valsa canker, was developed. One pair of genus-specific primers was designed based on the ribosomal DNA internal transcribed spacer conservative sequence of the Valsa genus and one pair of species-specific primers was designed based on the specific sequence of V. mali var. mali. The specificity of the genus-specific and species-specific primers was evaluated against 10 V. mali var. mali isolates, 10 V. mali var. pyri isolates, 4 isolates from closely related Valsa spp., and 8 isolates from fungal species that are commonly isolated from naturally infected apple bark tissue. A distinct band of 348 bp in length was detected in all V. mali var. mali isolates but not in other tested species and the V. mali var. pyri variety. The sensitivity of this assay was evaluated by serial dilutions of DNA extracted from V. mali var. mali pure cultures and apple bark tissues with or without visible symptoms. The results showed that the assay was able to detect as little as 100 fg of DNA in mycelial samples and apple bark tissues with visible symptoms, whereas the lowest detectable concentration was 10 pg of DNA in symptomless apple bark tissues. The efficiency of the nested PCR assay was compared with that of fungal isolation assays. All symptomless and symptomatic samples from which the pathogen was successfully isolated yielded a PCR product of the expected size. The detection rate of nested PCR for symptomless samples was 64.7%, which was much higher than the detection rate of 20.6% by fungal isolation. The PCR analysis of different symptomless tissues showed that the incidence of V. mali var. mali was different in different tissues of apple trees. The average incidence of V. mali var. mali was 89% in terminal buds, 71% in internodes, and 48% in bud scale scars. Moreover, the incidence of V. mali var. mali in nonsymptomatic tissues was higher in orchards where more trees were infected. Taken together, the assay developed in this study can be used for rapid and reliable detection of V. mali var. mali in tissues of apple trees with or without symptoms and also for monitoring the presence of the pathogen at an early stage of disease development.


MMP-2 and MMP-9 are prominent matrix metalloproteinases during atherosclerosis development in the Ldlr(-/-)Apob(100/100) mouse.
Matrix-degrading proteases capable of degrading components of the extracellular matrix may play an important role in development and progression of atherosclerotic lesions. In the present study, we used the Ldlr(-/-)Apob(100/100) mouse model, which has a plasma lipoprotein profile similar to that of humans with atherosclerosis, to study the expression of matrix metalloproteinases (MMPs) during early stages of atherosclerosis development. We analyzed the expression of 11 proteases and three protease inhibitors in 5- to 40-week-old Ldlr(-/-)Apob(100/100) mice. Expression and activity of MMP-2 and MMP-9 was increased in advanced atherosclerotic lesions followed by macrophage infiltration as shown by real-time PCR, gel-based and in situ zymography and immunohistochemistry. Expression of other investigated MMPs did not increase during disease progression. However, the mRNA expression of MMP-8 and MMP-13 was down-regulated, which could explain the relatively high amount of collagen observed in the vessels in this model. In conclusion, low proteolytic expression at early stages of atherogenesis and a limited repertoire of proteolytic enzymes were associated with the progression of atherosclerosis in Ldlr(-/-)Apob(100/100) mice. The study suggests that MMP-2 and MMP-9 are the main proteases involved in atherogenesis in this mouse model.


Serum cystatin C level is a useful marker for the evaluation of renal function in patients with cirrhotic ascites and normal serum creatinine levels.
Several studies suggested that serum cystatin C (CysC) is more useful than serum creatinine (Cr) for the assessment of renal function in patients with liver cirrhosis. This study evaluated the clinical significance of CysC in patients with cirrhotic ascites and normal Cr level. We enrolled patients with cirrhotic ascites and a normal serum Cr level (<1.2 mg/dL). GFR was measured by (99m)Tc-DTPA renal scan. Serum Cr, CysC, and Cr clearance (CCr) were measured on the same day. Significant renal impairment and severe renal impairment were defined as GFR <60 mL/min and GFR <30 mL/min, respectively. Eighty-nine patients with cirrhotic ascites were enrolled in the study (63 men and 26 women; age, 55±11 years). Forty-seven (52.8%) and 42 (47.2%) patients were in Child-Pugh grade B and C, respectively. Serum Cr and CysC levels and GFR were 0.8±0.2 mg/dL, 1.1±0.3 mg/L, and 73.4±25.5 mL/min, respectively. Significant and severe renal impairment were noted in 28 (31.5%) and 2 (2.2%) patients, respectively. GFR was well correlated with serum Cr, CysC, and e-GFR(MDRD), while it was not correlated with e-GFR(C&G). In multivariate analysis, only CysC was significantly correlated with GFR (β, 45.620; 95% CI, 23.042-68.198; P<0.001). Serum CysC level was the only independent predictor for significant renal impairment. Significant renal dysfunction was not rare in patients with cirrhotic ascites, even their serum Cr level is normal. Serum CysC is a useful marker for detecting significant renal dysfunction in these patients.


Dengue virus envelope protein domain I/II hinge determines long-lived serotype-specific dengue immunity.
The four dengue virus (DENV) serotypes, DENV-1, -2, -3, and -4, are endemic throughout tropical and subtropical regions of the world, with an estimated 390 million acute infections annually. Infection confers long-term protective immunity against the infecting serotype, but secondary infection with a different serotype carries a greater risk of potentially fatal severe dengue disease, including dengue hemorrhagic fever and dengue shock syndrome. The single most effective measure to control this threat to global health is a tetravalent DENV vaccine. To date, attempts to develop a protective vaccine have progressed slowly, partly because the targets of type-specific human neutralizing antibodies (NAbs), which are critical for long-term protection, remain poorly defined, impeding our understanding of natural immunity and hindering effective vaccine development. Here, we show that the envelope glycoprotein domain I/II hinge of DENV-3 and DENV-4 is the primary target of the long-term type-specific NAb response in humans. Transplantation of a DENV-4 hinge into a recombinant DENV-3 virus showed that the hinge determines the serotype-specific neutralizing potency of primary human and nonhuman primate DENV immune sera and that the hinge region both induces NAbs and is targeted by protective NAbs in rhesus macaques. These results suggest that the success of live dengue vaccines may depend on their ability to stimulate NAbs that target the envelope glycoprotein domain I/II hinge region. More broadly, this study shows that complex conformational antibody epitopes can be transplanted between live viruses, opening up similar possibilities for improving the breadth and specificity of vaccines for influenza, HIV, hepatitis C virus, and other clinically important viral pathogens.


Performance and return-to-sport after ACL reconstruction in NFL quarterbacks.
Anterior cruciate ligament (ACL) rupture is a significant injury in National Football League (NFL) quarterbacks. The purpose of this study was to determine (1) return-to-sport (RTS) rate in NFL quarterbacks following ACL reconstruction, (2) performance upon RTS, and (3) the difference in RTS and performance between players who underwent ACL reconstruction and controls. Thirteen quarterbacks (14 knees) who met inclusion criteria underwent ACL reconstruction while in the NFL. Matched controls were selected from the NFL during the same time span to compare and analyze age, body mass index (BMI), position, performance, and NFL experience. Student t tests were performed for analysis of within- and between-group variables. Bonferroni correction was used in the setting of multiple comparisons. Twelve quarterbacks (13 knees; 92%) were able to RTS in the NFL. Mean player age was 27.2±2.39 years. Mean career length in the NFL following ACL reconstruction was 4.85±2.7 years. Only 1 player needed revision ACL reconstruction. In both cases and controls, player performance was not significantly different from preinjury performance after ACL reconstruction (or index year in controls). There was also no significant performance difference between case and control quarterbacks following ACL reconstruction (or index year in controls). There is a high rate of RTS in the NFL following ACL reconstruction. In-game performance following ACL reconstruction was not significantly different from preinjury or from controls.


Enhancing palliative care in rural Australia: the residential aged care setting.
The delivery of palliative care in residential aged care communities is challenging, even more so in rural areas due to workforce ageing and shortages. The objectives of the present study were to: (i) assess the needs of, and quality of palliative care delivered to residents of 16 residential aged care facilities in rural southern Australia; and (ii) identify the needs of care staff to facilitate the delivery of quality palliative care. A cross-sectional survey of all residents, assessing the degree of functional limitation, stage of palliative care, and the presence of several quality indicators was conducted. Separate focus groups of care staff and relatives of residents sought information on the quality of care delivered, perceived strengths and weaknesses of the care delivered, and education and training needs. Quality palliative care in residential aged care facilities (RACFs) is hampered by workforce shortages, with low ratios of registered nurses, limited access to general practitioners after hours, and some communication difficulties. Some staff reported low confidence in technical and psychosocial aspects of care, especially for relatives. Relatives described mostly appropriate care, while acknowledging workload constraints. Most residents whose condition was unstable, deteriorating or terminal received advance care planning, though family expectations and unwillingness to discuss end-of-life care did tend to delay planning. Unstable residents with a reasonable prognosis were more likely to be transferred to hospital than terminally ill residents. Palliative care in participating RACFs appears to be adequate. Provision of targeted education for health care providers and implementation of protocols for advance care planning and end-of life care pathways will enhance this care.


Hnf4a deletion in the mouse kidney phenocopies Fanconi renotubular syndrome.
Different nephron tubule segments perform distinct physiological functions, collectively acting as a blood filtration unit. Dysfunction of the proximal tubule segment can lead to Fanconi renotubular syndrome (FRTS), with major symptoms such as excess excretion of water, glucose, and phosphate in the urine. It has been shown that a mutation in HNF4A is associated with FRTS in humans and that Hnf4a is expressed specifically in proximal tubules in adult rat nephrons. However, little is known about the role of Hnf4a in nephrogenesis. Here, we found that Hnf4a is expressed in both presumptive and differentiated proximal tubules in the developing mouse kidney. We show that Hnf4a is required for the formation of differentiated proximal tubules but is dispensable for the formation of presumptive proximal tubules. Furthermore, we show that loss of Hnf4a decreased the expression of proximal tubule-specific genes. Adult Hnf4a mutant mice presented with FRTS-like symptoms, including polyuria, polydipsia, glycosuria, and phosphaturia. Analysis of the adult Hnf4a mutant kidney also showed proximal tubule dysgenesis and nephrocalcinosis. Our results demonstrate the critical role of Hnf4a in proximal tubule development and provide mechanistic insight into the etiology of FRTS.


5'nucleotidase is sorted to the apical domain of hepatocytes via an indirect route.
In hepatocytes, all newly synthesized plasma membrane (PM) proteins so far studied arrive first at the basolateral domain; apically destined proteins are subsequently endocytosed and sorted to the apical domain via transcytosis. A mechanism for the sorting of newly synthesized glycophosphatidylinositol (GPI)-linked proteins has been proposed whereby they associate in lipid microdomains in the trans-Golgi network and then arrive at the apical domain directly. Such a mechanism poses a potential exception to the hepatocyte rule. We have used pulse-chase techniques in conjunction with subcellular fractionation to compare the trafficking of 5' nucleotidase (5NT), an endogenous GPI-anchored protein of hepatocytes, with two transmembrane proteins. Using a one-step fractionation technique to separate a highly enriched fraction of Golgi-derived membranes from ER and PM, we find that both 5NT and the polymeric IgA receptor (pIgAR) traverse the ER and Golgi apparatus with high efficiency. Using a method that resolves PM vesicles derived from the apical and basolateral domains, we find that 5NT first appears at the basolateral domain as early as 30 min of chase. However the subsequent redistribution to the apical domain requires > 3.5 h of chase to reach steady state. This rate of transcytosis is much slower than that observed for dipeptidylpeptidase IV, an apical protein anchored via a single transmembrane domain. We propose that the slow rate of transcytosis is related to the fact that GPI-linked proteins are excluded from clathrin-coated pits/vesicles, and instead must be endocytosed via a slower nonclathrin pathway.


Genetic regulation of delayed-type hypersensitivity responses to poly(LTyr,LGu)-poly(DLAla)--poly(LLys). I. Expression of the genetic defect at two phases of the immune process.
Delayed-type hypersensitivity (DTH) responses served in this study as an experimental model for the analysis of genetic regulations of T-cell responses. Educated irradiated cells from H-2b mice mediated responses in syngeneic recipients, whereas mice of the a, d, f, k, and s haplotypes were nonresponders to poly(LTyr,LGlu)-poly(DLAla)--poly(LLys)[(T,G)-A--L]. These results suggest that cell-mediated immune responsiveness to (T,G)-A--L is linked to the H-2 complex, as was shown for humoral responses. Educated irradiated T cells of F1 hybrids between high and low responders mediated DTH responses, which indicates that the gene(s) controlling the DTH responses is dominant. To analyze the genetic defect in DTH responses to (T,G)-A--L, we separated the T-cell activation phase from the effector phase that was determined in recipient mice. Two types of nonresponders were observed: (a) When lymphocytes of the a or k haplotypes were educated in a syngeneic environment and then transferred into hybrids between the parental (nonresponder x responder) F1 recipients, DTH responses could have been manifested. (b) On the other hand, no DTH responses could be mediated by transferring educated cells of the H-2s or H-2f origin into the appropriate F1 recipients. In addition, irradiated F1 cells that had been activated to (T,G)-A--L could not mediate DTH responses in both types of nonresponder recipients. These results suggest that T cells of H-2k or H-2a mice can be activated to generate DTH responses to (T,G)-A--L and that the defect in these mouse strains is expressed in another cell population needed for the manifestation of the DTH reaction in the recipient mice. In contrast, T cells of H-2s and H-2f origin cannot be activated to (T,G)-A--L and, thus, fail to manifest DTH responses.


Marital state, alcohol consumption, and liability to myocardial infarction, stroke, diabetes mellitus, or hypertension in men from Gothenburg.
Data from the Swedish personal number system were used to examine rates of admissions to hospital, particularly with myocardial infarction, stroke, diabetes mellitus, or hypertension, in relation to marital state and histories of admission with alcoholism in 77843 men from Gothenburg born in 1911-40. The number of admissions to hospital with the four diseases varied with marital state and history of admission with alcoholism, but by considering each subset from the total group it was possible to show that among alcoholic men no material variation in the number of admissions was evident. By contrast, among the non-alcoholic men those who were married were particularly prone to myocardial infarction but less likely to be admitted with stroke or diabetes mellitus than the unmarried or divorced men. The data show the value of common personal and health statistical numbering systems in generating epidemiological information beyond that obtainable from aggregated data without a record linkage technique.


Effective Surfactants Blend Concentration Determination for O/W Emulsion Stabilization by Two Nonionic Surfactants by Simple Linear Regression.
In this work, O/W emulsion sets were prepared by using different concentrations of two nonionic surfactants. The two surfactants, tween 80(HLB=15.0) and span 80(HLB=4.3) were used in a fixed proportions equal to 0.55:0.45 respectively. HLB value of the surfactants blends were fixed at 10.185. The surfactants blend concentration is starting from 3% up to 19%. For each O/W emulsion set the conductivity was measured at room temperature (25±2°), 40, 50, 60, 70 and 80°. Applying the simple linear regression least squares method statistical analysis to the temperature-conductivity obtained data determines the effective surfactants blend concentration required for preparing the most stable O/W emulsion. These results were confirmed by applying the physical stability centrifugation testing and the phase inversion temperature range measurements. The results indicated that, the relation which represents the most stable O/W emulsion has the strongest direct linear relationship between temperature and conductivity. This relationship is linear up to 80°. This work proves that, the most stable O/W emulsion is determined via the determination of the maximum R² value by applying of the simple linear regression least squares method to the temperature-conductivity obtained data up to 80°, in addition to, the true maximum slope is represented by the equation which has the maximum R² value. Because the conditions would be changed in a more complex formulation, the method of the determination of the effective surfactants blend concentration was verified by applying it for more complex formulations of 2% O/W miconazole nitrate cream and the results indicate its reproducibility.


Neuro-ophthalmology of degenerative neurological disorders.
A number of degenerative disorders of the nervous system are associated with visual or ocular motor disturbances. Over recent years, these problems have attracted a great deal of interest because they may aid diagnosis and also improve our understanding of pathophysiology. In this review, we discuss both of these aspects of the study of degenerative conditions and attempt to demonstrate the clinical significance of a number of new findings.


Recurrent disk versus scar in the postoperative patient: the role of computed tomography (CT)/diskography and CT/myelography.
The purpose of this study was to compare computed tomography (CT)/myelography and CT/diskography images, in a given patient, as a method of distinguishing postoperative fibrosis from recurrent herniated disk material. The study population consisted of 20 patients who had undergone lumbar diskectomy and subsequently developed recurrent radicular pain. All patients underwent CT/myelography and CT/diskography, each procedure performed within 72 h of the other. Comparison of transaxial images from CT/myelography and CT/diskography at a given disk space level yielded the following results: in 12 patients the extradural mass seen via CT/myelography corresponded entirely to the contrast-filled disk fragment seen via CT/diskography (recurrent herniated disk); in three patients the extradural mass seen via CT/myelography was larger than the disk fragment seen via CT/diskography (recurrent herniated disk and fibrosis); and in five patients CT/diskography images appeared normal, but CT/myelography showed an extradural mass (fibrosis). Fifteen patients underwent surgical reexploration with the following results: in three of three patients, the suspicion of recurrent herniated disk and fibrosis by radiologic evaluation was confirmed by surgical reexploration; in nine of 12 patients, solely recurrent herniated disk shown by radiologic criteria was similarly confirmed. The remaining five patients presumed to have fibrosis by radiologic criteria were treated nonoperatively.


Randomized, split-body, single-blinded clinical trial of topical broccoli sprout extract: Assessing the feasibility of its use in keratin-based disorders.
Epidermolysis bullosa simplex is a skin-blistering disorder caused by mutations in keratin (K)14 or K5. Treatment with nuclear factor (erythroid-derived 2)-like 2 inducer sulforaphane ameliorated skin blistering in Krt14-null mice, correlating with induction of K17. To be therapeutically useful for epidermolysis bullosa simplex, topical broccoli sprout extract (BSE), enriched for sulforaphane, would ideally induce the expression of homologous keratins (eg, K6, K17, K16) in the basal layer of human epidermis without impacting expression of defective keratins (K5/K14). The purpose of this 1-week, randomized, split-body, single-blinded, placebo-controlled trial was to assess the impact of BSE on keratin expression. Five subjects (34-71 years old) applied BSE (500 nmol of sulforaphane/mL) or vehicle alone to the inner aspect of the arm daily. Expression of keratin, nuclear factor (erythroid-derived 2)-like 2, and other markers was assessed using reverse transcription-polymerase chain reaction and indirect immunofluorescence. One subject (age 71 years) was excluded a posteriori because of poor tissue quality. Topical BSE activated nuclear factor (erythroid-derived 2)-like 2 and up-regulated K17 in the epidermis of all subjects, had variable effects on K16 and K6 expression, and did not alter expression of K14 or K5. Small sample size is a limitation. BSE represents an attractive therapeutic candidate for K14-associated epidermolysis bullosa simplex.


Effect of some antioxidants on sarcoplasmic reticulum Ca2+-ATPase activity from rabbit skeletal muscle.
Effects of phenolic antioxidants, synthetized (trolox, pyridoindole stobadine) and plant extracts (EGb 761 and Pycnogenol) were investigated on the activity of Ca(2+)-ATPase from sarcoplasmic reticulum (SR) of rabbit skeletal muscle to examine their potency to modulate the activity of this calcium level regulating enzyme. SR vesicles and pure Ca(2+)-ATPase were isolated by ultracentrifugation. Ca(2+)-ATPase activity was measured spectrophotometrically by enzyme-coupled assay. Pycnogenol (Pyc) significantly decreased the activity of Ca(2+)-ATPase incorporated into SR vesicles as well as the activity of purified Ca(2+)-ATPase, the latter with respect to both enzyme substrates, Ca(2+) and ATP. Trolox, stobadine and EGb 761 did not influence significantly the activity SR- vesicle incorporated or pure Ca(2+)-ATPase, the latter with respect to either substrate, in spite of alterations of kinetic parameters in some cases. The decrease of SR Ca(2+)-ATPase activity induced by Pyc may be associated with its proapoptotic and anticancerogenic properties.


Integrative mechanisms of blood pressure regulation in humans and rats: cross-species similarities.
As our understanding of the importance of individualized medicine continues to grow, the clinical relevance of interindividual variability in hemodynamic variables is receiving increasing attention. However, it is not known whether the rat, which is often used for studies of cardiovascular regulation, exhibits similar interindividual variability. In the present study, we evaluated whether the magnitude of interindividual variability in cardiac output (CO) and total peripheral resistance (TPR) was similar in humans and in rats. We assessed interindividual variability of mean arterial pressure (MAP), CO, and TPR during control conditions in normotensive humans (n = 40) and during normotension and deoxycorticosterone acetate-salt hypertension in Sprague-Dawley rats (n = 16). Humans and rats showed marked interindividual variability in CO and TPR but low variability in MAP. During deoxycorticosterone acetate-salt hypertension, CO was maintained, but TPR was elevated compared with the baseline period. We conclude that the magnitudes of interindividual variability of MAP, CO, and TPR are quantitatively similar in humans and rats, providing support for the relevance of this variability in both species and suggesting that studies in rats could be designed to address questions specific to individualized medicine in hypertension.


Suboccipital decompression during posterior cranial vault remodeling for selected cases of Chiari malformations associated with craniosynostosis.
The optimal management of Chiari malformations in the setting of craniosynostosis is not well established. In this report the authors describe their outcomes with the combined technique of simultaneous suboccipital decompression (SOD) during posterior cranial vault remodeling (PCVR). A retrospective review was performed of all patients undergoing PCVR and simultaneous SOD. Demographic data, diagnosis, imaging studies, operative intervention, and clinical follow-up were evaluated. Thirty-four patients were identified as having undergone a simultaneous PCVR/SOD for Chiari malformation associated with craniosynostosis. Eighty-eight percent of these patients had syndromic, multisutural craniosynostosis, and the remaining patients had unilateral lambdoid craniosynostosis. There were no postoperative complications as a direct result from this combined procedure. Two patients required a subsequent direct approach for decompression of the Chiari malformation. The interval between these subsequent surgeries was 3 years and 19 months. Chiari malformations are commonly associated with syndromic, complex craniosynostosis and isolated lambdoid craniosynostosis. In appropriately selected patients, a combined posterior cranial vault enlargement and SOD of the foramen magnum is associated with a low complication rate and appears to be an effective procedure.


Can comodulation masking release occur when frequency changes could promote perceptual segregation of the on-frequency and flanking bands?
A common characteristic of natural sounds is that the level fluctuations in different frequency regions are coherent. The ability of the auditory system to use this comodulation is shown when a sinusoidal signal is masked by a masker centred at the signal frequency (on-frequency masker, OFM) and one or more off-frequency components, commonly referred to as flanking bands (FBs). In general, the threshold of the signal masked by comodulated masker components is lower than when masked by masker components with uncorrelated envelopes or in the presence of the OFM only. This effect is commonly referred to as comodulation masking release (CMR). The present study investigates if CMR is also observed for a sinusoidal signal embedded in the OFM when the centre frequencies of the FBs are swept over time with a sweep rate of one octave per second. Both a common change of different frequencies and comodulation could serve as cues to indicate which of the stimulus components originate from one source. If the common fate of frequency components is the stronger binding cue, the sweeping FBs and the OFM with a fixed centre frequency should no longer form one auditory object and the CMR should be abolished. However, psychoacoustical results with normal-hearing listeners show that a CMR is also observed with sweeping components. The results are consistent with the hypothesis of wideband inhibition as the underlying physiological mechanism, as the CMR should only depend on the spectral position of the flanking bands relative to the inhibitory areas (as seen in physiological recordings using stationary flanking bands). Preliminary physiological results in the cochlear nucleus of the Guinea pig show that a correlate of CMR can also be found at this level of the auditory pathway with sweeping flanking bands.


Cerebral small vessel disease: Capillary pathways to stroke and cognitive decline.
Cerebral small vessel disease (SVD) gives rise to one in five strokes worldwide and constitutes a major source of cognitive decline in the elderly. SVD is known to occur in relation to hypertension, diabetes, smoking, radiation therapy and in a range of inherited and genetic disorders, autoimmune disorders, connective tissue disorders, and infections. Until recently, changes in capillary patency and blood viscosity have received little attention in the aetiopathogenesis of SVD and the high risk of subsequent stroke and cognitive decline. Capillary flow patterns were, however, recently shown to limit the extraction efficacy of oxygen in tissue and capillary dysfunction therefore proposed as a source of stroke-like symptoms and neurodegeneration, even in the absence of physical flow-limiting vascular pathology. In this review, we examine whether capillary flow disturbances may be a shared feature of conditions that represent risk factors for SVD. We then discuss aspects of capillary dysfunction that could be prevented or alleviated and therefore might be of general benefit to patients at risk of SVD, stroke or cognitive decline.


Biosynthesis of crocacin involves an unusual hydrolytic release domain showing similarity to condensation domains.
The crocacins are potent antifungal and cytotoxic natural compounds from myxobacteria of the genus Chondromyces. Although total synthesis approaches have been reported, the molecular and biochemical basis guiding the formation of the linear crocacin scaffold has remained unknown. Along with the identification and functional analysis of the crocacin biosynthetic gene cluster from Chondromyces crocatus Cm c5, we here present the identification and biochemical characterization of an unusual chain termination domain homologous to condensation domains responsible for hydrolytic release of the product from the assembly line. In particular, gene inactivation studies and in vitro experiments using the heterologously produced domain CroK-C2 confirm this surprising role giving rise to the linear carboxylic acid. Additionally, we determined the kinetic parameters of CroK-C2 by monitoring hydrolytic cleavage of the substrate mimic N-acetylcysteaminyl-crocacin B using an innovative high-performance liquid chromatography mass spectrometry-based assay.


Inclusion complexation of artemisinin with alpha-, beta-, and gamma-cyclodextrins.
The present study was conducted to investigate the inclusion complexation of artemisinin (ART) with natural cyclodextrins (CyD), namely alpha-, beta-, and gamma-CyDs with the aim of improving its solubility and dissolution rate. Complex formation in aqueous solution and solid state was studied by solubility analysis, dissolution, and thermal analysis. Solubility diagrams indicated that the complexation of ART and the three CyDs occurred at a molar ratio of 1:1, and showed a remarkable increase in ART solubility. Moreover, the thermodynamic parameters calculated by using the van't Hoff equation revealed that the complexation process was associated with negative enthalpy of formation and occurred spontaneously. The complexation capability of CyDs with ART increased in the order of alpha- < gamma- < beta-CyDs and could be ascribed to the structural compatibility between the molecular size of ART and the diameter of the CyD cavities. Dissolution profiles of the three complexes demonstrated an increased rate and extent of dissolution compared with those of their respective physical mixtures and a commercial preparation. In solid-state analysis, using differential scanning calorimetry, the gamma-CyD was capable of complexing the highest percentage of ART, followed by beta- and alpha-CyDs. The respective estimated percentage of ART complexed by the CyDs were 85%, 40%, and 12%.


Systemic aminoglycoside treatment in rodent models of retinitis pigmentosa.
We studied the potential of systemically administered aminoglycosides as a therapy for retinal degeneration resulting from premature termination codon (PTC) mutations. Aminoglycosides were systemically delivered to two rodent models of retinal degeneration: a transgenic rat model of dominant disease caused by a PTC in rhodopsin (S334ter); and a mouse model of recessive disease (rd12) caused by a PTC in the retinoid isomerase Rpe65. Initial luciferase reporter assays were undertaken to measure the efficiency of gentamicin-induced read-through in vitro. These experiments indicated that gentamicin treatment induced on average a 5.3% extra read-through of the S334ter PTC in vitro, but did not affect the rd12 PTC. Beginning at postnatal day 5, animals received daily subcutaneous injections of gentamicin or geneticin at a range of doses. The effect of the treatment on retinal degeneration was examined by histopathology and electroretinography (ERG). Systemic treatment with aminoglycoside significantly increased the number of surviving photoreceptors in the S334ter rat model over several weeks of treatment, but was not effective in slowing the retinal degeneration in the rd12 mouse model. Similarly, ERG recordings indicated better preservation of retinal function in the treated S334ter rats, but no difference was observed in the rd12 mice. Daily subcutaneous injection of 12.5mug/g gentamicin was the only regimen that inhibited retinal degeneration without apparent adverse systemic side effects. Reduced effectiveness beyond postnatal day 50 correlated with reduced ocular penetration of drug as seen in gentamicin-Texas red (GTTR) conjugation experiments. We conclude that, in the rat model, an approximately 5% reduction of abnormal truncated protein is sufficient to enhance photoreceptor survival. Such a change in truncated protein is consistent with beneficial effects seen when aminoglycosides has been used in other, non-ocular animal models. In the rd12 mouse, lack of efficacy was seen despite this particular PTC being theoretically more sensitive to aminoglycoside modification. We conclude that aminoglycoside read-through of PTCs in vitro and in vivo cannot be predicted just from genomic context. Because there is considerable genetic heterogeneity amongst retinal degenerations, pharmacologic therapies that are not gene-specific have significant appeal. Our findings suggest that if adverse issues such as systemic toxicity and limited ocular penetration can be overcome, small molecule therapeutics, such as aminoglycosides, which target classes of mutation could hold considerable potential as therapies for retinal disease.


The risk of virologic failure decreases with duration of HIV suppression, at greater than 50% adherence to antiretroviral therapy.
We hypothesized that the percent adherence to antiretroviral therapy necessary to maintain HIV suppression would decrease with longer duration of viral suppression. Eligible participants were identified from the REACH cohort of marginally housed HIV infected adults in San Francisco. Adherence to antiretroviral therapy was measured through pill counts obtained at unannounced visits by research staff to each participant's usual place of residence. Marginal structural models and targeted maximum likelihood estimation methodologies were used to determine the effect of adherence to antiretroviral therapy on the probability of virologic failure during early and late viral suppression. A total of 221 subjects were studied (median age 44.1 years; median CD4+ T cell nadir 206 cells/mm(3)). Most subjects were taking the following types of antiretroviral regimens: non-nucleoside reverse transcriptase inhibitor based (37%), ritonavir boosted protease inhibitor based (28%), or unboosted protease inhibitor based (25%). Comparing the probability of failure just after achieving suppression vs. after 12 consecutive months of suppression, there was a statistically significant decrease in the probability of virologic failure for each range of adherence proportions we considered, as long as adherence was greater than 50%. The estimated risk difference, comparing the probability of virologic failure after 1 month vs. after 12 months of continuous viral suppression was 0.47 (95% CI 0.23-0.63) at 50-74% adherence, 0.29 (CI 0.03-0.50) at 75-89% adherence, and 0.36 (CI 0.23-0.48) at 90-100% adherence. The risk of virologic failure for adherence greater than 50% declines with longer duration of continuous suppression. While high adherence is required to maximize the probability of durable viral suppression, the range of adherence capable of sustaining viral suppression is wider after prolonged periods of viral suppression.


A Laplace mixture model for identification of differential expression in microarray experiments.
Microarrays have become an important tool for studying the molecular basis of complex disease traits and fundamental biological processes. A common purpose of microarray experiments is the detection of genes that are differentially expressed under two conditions, such as treatment versus control or wild type versus knockout. We introduce a Laplace mixture model as a long-tailed alternative to the normal distribution when identifying differentially expressed genes in microarray experiments, and provide an extension to asymmetric over- or underexpression. This model permits greater flexibility than models in current use as it has the potential, at least with sufficient data, to accommodate both whole genome and restricted coverage arrays. We also propose likelihood approaches to hyperparameter estimation which are equally applicable in the Normal mixture case. The Laplace model appears to give some improvement in fit to data, though simulation studies show that our method performs similarly to several other statistical approaches to the problem of identification of differential expression.


Passion fruit peel extract attenuates bleomycin-induced pulmonary fibrosis in mice.
Idiopathic pulmonary fibrosis is a progressive fatal lung disease characterized by excessive collagen deposition, with no effective treatments. We investigated the efficacy of natural products with high anti-inflammatory activity, such as passion fruit peel extract (PFPE), in a mouse model of bleomycin-induced pulmonary fibrosis (PF). C57BL/6J mice were subjected to a single intratracheal instillation of bleomycin to induce PF. Daily PFPE treatment significantly reduced loss of body mass and mortality rate in mice compared with those treated with bleomycin. While bleomycin-induced PF resulted in elevated total numbers of inflammatory cells, macrophages, lymphocytes, and neutrophils in bronchoalveolar lavage fluid on both days 7 and 21, PFPE administration significantly attenuated these phenomena compared with bleomycin group. On day 7, the decreased superoxide dismutase and myeloperoxidase activities observed in the bleomycin group were significantly restored with PFPE treatment. On day 21, enhanced hydroxyproline deposition in the bleomycin group was also suppressed by PFPE administration. PFPE treatment significantly attenuated extensive inflammatory cell infiltration and accumulation of collagen in lung tissue sections of bleomycin-induced mice on days 7 and 21, respectively. Our results indicate that administration of PFPE decreased bleomycin-induced PF because of anti-inflammatory and antioxidant activities.


[Awareness, treatment, and control of arterial hypertension: Pró-Saúde study, Brazil].
To evaluate the prevalence, awareness, treatment, and control of hypertension in a working-age population in Brazil. This project is part of a prospective study with employees at a university in Rio de Janeiro (Pró-Saúde Study). Based on standardized measurements of blood pressure and on the history of diagnosis and treatment of hypertension, we estimated the prevalence of hypertension in 2 384 participants from the baseline population (1999 to 2001) and determined what percentage, according to sex and age, were aware of the diagnosis of arterial hypertension, treated this condition, and had their disease controlled with anti-hypertensive drugs. Of the population under study, 704 people (29.6%) were classified as hypertensive; of these, 573 (81.5%) were aware of their condition (88.6% of females and 71.6% of males). Of those who were aware, 445 (77.8%; 88.8% of females and 59.1% of males) were being treated with anti-hypertensive drugs. In turn, of the hypertensive subjects being treated for this condition, 287 (60.1%; 61.5% of females and 57.4% of males) were controlled. Awareness and treatment rates were lower and control was higher in those in the lowest age range. In the study population, awareness, treatment, and control of hypertension were higher than has been reported by most national and international studies. Considering the fact that most individuals were aware of and treated their condition, a higher rate of hypertension control was expected. Adequate control of hypertension, especially among men, must be a priority of health programs and services.


Allergic contact dermatitis in children and young adults.
We evaluated the results of a 5-year period of epicutaneous testing in children and teenagers younger than 16 years of age. In an overall-tested population of 2671 persons, 67 were younger than 16 years. In 16 (23.8%) of these 67 children, a positive epicutaneous test was considered to be clinically relevant for the diagnosis allergic contact dermatitis. From the total of 44 positive tests, the most common allergen was nickel sulfate (18%), followed by balsam of Peru, Carba mix, colophony, and fragrance mix (6% each). As our test population consisted of only a few children, no conclusions were drawn in respect to children younger than 10 years. Despite our careful selection of children with allergic contact dermatitis, the prevalence of positive epicutaneous tests was no different than that reported for unselected populations of children.


Microbial stress priming: a meta-analysis.
Microbes have to cope with complex and dynamic environments, making it likely that anticipatory responses provide fitness benefits. Mild, previous stressors can prepare microbes (stress priming) to further and potentially damaging stressors (triggering). We here quantitatively summarize the findings from over 250 trials of 34 studies including bacteria and fungi, demonstrating that priming to stress has a beneficial impact on microbial survival. In fact, survival of primed microbes was about 10-fold higher compared with that in non-primed microbes. Categorical moderators related to microbial taxonomy and the kind of stress applied as priming or as triggering revealed significant differences of priming effect size among 14 different microbial species, 6 stress categories and stressor combination. We found that priming by osmotic, physiological and temperature stress had the highest positive effect sizes on microbial response. Cross-protection was evident for physiological, temperature and pH stresses. Microbes are better prepared against triggering by oxidative, temperature and osmotic stress. Our finding of an overall positive mean effect of priming regardless of the microbial system and particular stressor provides unprecedentedly strong evidence of the broad ecological significance of microbial stress priming. These results further suggest that stress priming may be an important factor in shaping microbial communities.


Doppleron-induced stimulated reflection.
In a gaseous system, energy can be transferred from an intense incoming beam to a counterpropagating probe beam at the same frequency. The effect may be interpreted in terms of Doppleron processes and arises because the medium undergoes interaction with beams of different effective frequencies owing to the Doppler effect.


The visible skeleton 2.0: phenotyping of cartilage and bone in fixed vertebrate embryos and foetuses based on X-ray microCT.
For decades, clearing and staining with Alcian Blue and Alizarin Red has been the gold standard to image vertebrate skeletal development. Here, we present an alternate approach to visualise bone and cartilage based on X-ray microCT imaging, which allows the collection of genuine 3D data of the entire developing skeleton at micron resolution. Our novel protocol is based on ethanol fixation and staining with Ruthenium Red, and efficiently contrasts cartilage matrix, as demonstrated in whole E16.5 mouse foetuses and limbs of E14 chicken embryos. Bone mineral is well preserved during staining, thus the entire embryonic skeleton can be imaged at high contrast. Differences in X-ray attenuation of ruthenium and calcium enable the spectral separation of cartilage matrix and bone by dual energy microCT (microDECT). Clearing of specimens is not required. The protocol is simple and reproducible. We demonstrate that cartilage contrast in E16.5 mouse foetuses is adequate for fast visual phenotyping. Morphometric skeletal parameters are easily extracted. We consider the presented workflow to be a powerful and versatile extension to the toolkit currently available for qualitative and quantitative phenotyping of vertebrate skeletal development.


Function of the heterocercal tail in white sturgeon: flow visualization during steady swimming and vertical maneuvering.
Basal ray-finned fishes possess a heterocercal tail in which the dorsal lobe containing the extension of the vertebral column is longer than the ventral lobe. Clarifying the function of the heterocercal tail has proved elusive because of the difficulty of measuring the direction of force produced relative to body position in the aquatic medium. We measured the direction of force produced by the heterocercal tail of the white sturgeon (Acipenser transmontanus) by visualizing flow in the wake of the tail using digital particle image velocimetry (DPIV) while simultaneously recording body position and motion using high-speed video. To quantify tail function, we measured the vertical body velocity, the body angle and the path angle of the body from video recordings and the vortex ring axis angle and vortex jet angle from DPIV recordings of the wake downstream from the tail. These variables were measured for sturgeon exhibiting three swimming behaviors at 1.2 L s(-)(1), where L is total body length: rising through the water column, holding vertical position, and sinking through the water column. For vertical body velocity, body angle and path angle values, all behaviors were significantly different from one another. For vortex ring axis angle and vortex jet angle, rising and holding behavior were not significantly different from each other, but both were significantly different from sinking behavior. During steady horizontal swimming, the sturgeon tail generates a lift force relative to the path of motion but no rotational moment because the reaction force passes through the center of mass. For a rising sturgeon, the tail does not produce a lift force but causes the tail to rotate ventrally in relation to the head since the reaction force passes ventral to the center of mass. While sinking, the direction of the fluid jet produced by the tail relative to the path of motion causes a lift force to be created and causes the tail to rotate dorsally in relation to the head since the reaction force passes dorsal to the center of mass. These data provide evidence that sturgeon can actively control the direction of force produced by their tail while maneuvering through the water column because the relationship between vortex jet angle and body angle is not constant.


Younger hepatocellular carcinoma patients have better prognosis after percutaneous radiofrequency ablation therapy.
To evaluate the impact of age on the prognosis of patients with hepatocellular carcinoma (HCC) undergoing percutaneous radiofrequency ablation (RFA). Whether age plays an important role in the outcomes of HCC after RFA remains controversial. Two hundred fifty-eight consecutive treatment naive HCC patients who underwent RFA were enrolled. Patients aged ≤ 65 years (n = 100) were defined as the younger group and those aged > 65 years (n = 158) were the elderly group. Their clinicopathologic features and prognosis were compared. Younger patients had a higher male-to-female ratio, higher prevalence of hepatitis B virus, and smaller tumor size than elder patients. After median follow-up of 28.5 ± 18.7 months, 45 patients died. The cumulative 5-year survival rates were 81.3% and 65.4% in younger and elder HCC patients, respectively (P = 0.008). Multivariate analysis disclosed that age > 65 years, serum albumin level ≤ 3.7 g/dL, prothrombin time international normalized ratio > 1.1, α-fetoprotein (AFP) > 20 ng/mL, and no antiviral therapy after RFA were independent risk factors associated with poor overall survival. Besides, there were 163 patients with tumor recurrence after RFA. Multivariate analysis showed that age > 65 years, platelet count ≤ 10/mm, AFP > 20 ng/mL, multinodularity, and tumor size > 2 cm were the independent risk factors predicting recurrence. Both liver functional reserve (serum albumin level, prothrombin time international normalized ratio, platelet count, and antiviral therapy) and tumor factors (tumor size, number, and AFP level) were crucial in determining post-RFA prognosis in HCC patients. Moreover, younger HCC patients have better overall survival and lower recurrence rate after RFA compared with elder patients.


Cell mass and cell cycle dynamics of an asynchronous budding yeast population: experimental observations, flow cytometry data analysis, and multi-scale modeling.
Despite traditionally regarded as identical, cells in a microbial cultivation present a distribution of phenotypic traits, forming a heterogeneous cell population. Moreover, the degree of heterogeneity is notably enhanced by changes in micro-environmental conditions. A major development in experimental single-cell studies has taken place in the last decades. It has however not been fully accompanied by similar contributions within data analysis and mathematical modeling. Indeed, literature reporting, for example, quantitative analyses of experimental single-cell observations and validation of model predictions for cell property distributions against experimental data is scarce. This study focuses on the experimental and mathematical description of the dynamics of cell size and cell cycle position distributions, of a population of Saccharomyces cerevisiae, in response to the substrate consumption observed during batch cultivation. The good agreement between the proposed multi-scale model (a population balance model [PBM] coupled to an unstructured model) and experimental data (both the overall physiology and cell size and cell cycle distributions) indicates that a mechanistic model is a suitable tool for describing the microbial population dynamics in a bioreactor. This study therefore contributes towards the understanding of the development of heterogeneous populations during microbial cultivations. More generally, it consists of a step towards a paradigm change in the study and description of cell cultivations, where average cell behaviors observed experimentally now are interpreted as a potential joint result of various co-existing single-cell behaviors, rather than a unique response common to all cells in the cultivation.


Functional form estimation using oblique projection matrices for LS-SVM regression models.
Kernel regression models have been used as non-parametric methods for fitting experimental data. However, due to their non-parametric nature, they belong to the so-called "black box" models, indicating that the relation between the input variables and the output, depending on the kernel selection, is unknown. In this paper we propose a new methodology to retrieve the relation between each input regressor variable and the output in a least squares support vector machine (LS-SVM) regression model. The method is based on oblique subspace projectors (ObSP), which allows to decouple the influence of input regressors on the output by including the undesired variables in the null space of the projection matrix. Such functional relations are represented by the nonlinear transformation of the input regressors, and their subspaces are estimated using appropriate kernel evaluations. We exploit the properties of ObSP in order to decompose the output of the obtained regression model as a sum of the partial nonlinear contributions and interaction effects of the input variables, we called this methodology Nonlinear ObSP (NObSP). We compare the performance of the proposed algorithm with the component selection and smooth operator (COSSO) for smoothing spline ANOVA models. We use as benchmark 2 toy examples and a real life regression model using the concrete strength dataset from the UCI machine learning repository. We showed that NObSP is able to outperform COSSO, producing stable estimations of the functional relations between the input regressors and the output, without the use of prior-knowledge. This methodology can be used in order to understand the functional relations between the inputs and the output in a regression model, retrieving the physical interpretation of the regression models.


Identification of soluble TREM-2 in the cerebrospinal fluid and its association with multiple sclerosis and CNS inflammation.
Triggering receptor expressed on myeloid cells 2 (TREM-2) is a membrane-bound receptor expressed by microglia and macrophages. Engagement of TREM-2 on these cells has been reported to reduce inflammatory responses and, in microglial cells, to promote phagocytosis. TREM-2 function is critical within the CNS, as its genetic deficiency in humans causes neurodegeneration with myelin and axonal loss. Blockade of TREM-2 worsened the mouse model for multiple sclerosis. In the present study, a soluble form of TREM-2 protein has been identified by immunoprecipitation and by ELISA. Soluble TREM-2 protein (sTREM-2) was detected in human CSF, and was compared among subjects with relapsing-remitting multiple sclerosis (RR-MS; n = 52), primary progressive multiple sclerosis (PP-MS; n = 21), other inflammatory neurologic diseases (OIND; n = 19), and non-inflammatory neurologic diseases (NIND; n = 41). Compared to NIND subjects, CSF sTREM-2 levels were significantly higher in RR-MS (P = 0.004 by ANOVA) and PP-MS (P < 0.001) subjects, as well as in OIND (P < 0.001) subjects. In contrast, levels of sTREM-2 in blood did not differ among the groups. Furthermore, TREM-2 was detected on a subset of CSF monocytes by flow cytometry, and was also highly expressed on myelin-laden macrophages in eight active demyelinating lesions from four autopsied multiple sclerosis subjects. The elevated levels of sTREM-2 in CSF of multiple sclerosis patients may inhibit the anti-inflammatory function of the membrane-bound receptor suggesting sTREM-2 to be a possible target for future therapies.


Multimolecular characteristics of cell-death related hub genes in human cancers: a comprehensive pan-cancer analysis.
Failure of the normal process of cell death pathways contributes to the defection of immune systems and the occurrence of cancers. The key genes, the multimolecular mechanisms, and the immune functions of these genes in pan-cancers remain unclear. Using online databases of The Cancer Genome Atlas, GEPIA2, TISIDB, HPA, Kaplan-Meier Plotter, PrognoScan, cBioPortal, GSCALite, TIMER, and Sangerbox, we identified the key genes from the six primary cell death-related pathways and performed a comprehensive analysis to investigate the multimolecular characteristics and immunological functions of the hub genes in 33 human cancers. We identified five hub genes in the six primary cell death-related pathways (JUN, NFKB1, CASP3, PARP1, and TP53). We found that CASP3, PARP1, and TP53 were overexpressed in 28, 23, and 27 cancers. The expression of the five genes was associated with the development and prognosis of many cancers. Particularly, JUN, NFKB1, CASP3, and TP53 have prognostic values in Brain Lower Grade Glioma (LGG), while PARP1 and CASP3 could predict the survival outcomes in Adrenocortical carcinoma (ACC). In addition, an extensive association between five genes' expression, DNA methylation, and tumor-immune system interactions was noticed. The five cell death-related hub genes could function as potential biomarkers for various cancers, particularly LGG and ACC. The immunological function analysis of the five genes also proposes new targets for developing immunosuppressants and improving the immunotherapy efficacy of cancers. However, further extensive clinical and experimental research are required to validate their clinical values.


Integration of proteomics and metabolomics reveals promotion of proliferation by exposure of bisphenol S in human breast epithelial MCF-10A cells.
Bisphenol S (BPS) has been reported to have similar estrogenic effects as bisphenol A (BPA). Considering the endocrine disrupting effects of BPS, in this study, we investigated the effects of BPS exposure on normal human breast epithelial cell line MCF-10A by using mass spectrometry (MS)-based metabolomics and quantitative proteomics. We found that exposure to BPS for 24 h altered the proliferation of MCF-10A cells in a hormetic manner with the highest proliferation rate at the dosage of 1 μM. A total of 200 proteins were identified to be significantly changed by 1 μM of BPS exposure. The upregulation of epidermal growth factor receptor (EGFR) and Ras/mTOR-related proteins implied that EGFR-mediated pathways were involved in BPS-induced proliferation of MCF-10A cells. In addition, several proliferation-related protein markers were found to be elevated, such as MKI67 and CDH1, further indicating the promotion of proliferation by low dose of BPS exposure. Besides, 35 endogenous metabolites were found to be significantly changed. The joint pathway analysis of the altered metabolites and proteins suggested changes in pathways of tricarboxylic acid (TCA) cycle, purine metabolism, pyruvate metabolism and lipid metabolism, which were involved in sustaining cell proliferation and cellular signal transduction. Taken together, this study provides insights into the effects and the potential mechanisms of BPS on estrogen receptor α-negative normal breast cell line MCF-10A, broadening our knowledge about the risk of using BPS as the alternative of BPA.


Wound healing dressings and drug delivery systems: a review.
The variety of wound types has resulted in a wide range of wound dressings with new products frequently introduced to target different aspects of the wound healing process. The ideal dressing should achieve rapid healing at reasonable cost with minimal inconvenience to the patient. This article offers a review of the common wound management dressings and emerging technologies for achieving improved wound healing. It also reviews many of the dressings and novel polymers used for the delivery of drugs to acute, chronic and other types of wound. These include hydrocolloids, alginates, hydrogels, polyurethane, collagen, chitosan, pectin and hyaluronic acid. There is also a brief section on the use of biological polymers as tissue engineered scaffolds and skin grafts. Pharmacological agents such as antibiotics, vitamins, minerals, growth factors and other wound healing accelerators that take active part in the healing process are discussed. Direct delivery of these agents to the wound site is desirable, particularly when systemic delivery could cause organ damage due to toxicological concerns associated with the preferred agents. This review concerns the requirement for formulations with improved properties for effective and accurate delivery of the required therapeutic agents. General formulation approaches towards achieving optimum physical properties and controlled delivery characteristics for an active wound healing dosage form are also considered briefly.


Development and behaviour of low-birthweight term infants at 8 years in northeast Brazil: a longitudinal study.
To investigate the development and behaviour of low-birthweight (LBW) term infants compared with matched term infants of appropriate birthweight (ABW). Two parallel cohorts of LBW infants (1500-2499 g) and ABW controls (3000-3499 g) were enrolled at birth in northeast Brazil. At 8 y, 164 children were assessed using clinical and psychological tests. The LBW group had lower IQ scores than ABW children on the Weschler Intelligence Scale for Children; differences were larger on the performance (5 points, p = 0.04) than the verbal scale (3 points, p = 0.11). LBW children had poorer dynamic balance (p = 0.03) and eye-hand coordination (p = 0.02), but better selective attention (p = 0.02). Hyperactivity and conduct problems were common in both groups, but fewer LBW children had peer problems (p = 0.04). After controlling for social background, IQ was not significantly associated with birthweight (p = 0.10). Significant determinants were maternal education, home stimulation and type of school attended. The effect of birthweight on coordination and selective attention remained significant. Birth head circumference and growth in head size in the first 6 mo had independent effects on IQ. After controlling for social background, LBW term infants did not differ from ABW infants in cognition at school age. Head circumference at birth and 6 mo was a better predictor than birthweight of IQ.


A meta-analysis of the yield of capsule endoscopy compared to other diagnostic modalities in patients with non-stricturing small bowel Crohn's disease.
Capsule endoscopy (CE) allows for direct evaluation of the small bowel mucosa in patients with Crohn's disease (CD). A number of studies have revealed significantly improved yield for CE over other modalities for the diagnosis of CD, but as sample sizes have been small, the true degree of benefit is uncertain. Additionally, it is not clear whether patients with a suspected initial presentation of CD and those with suspected recurrent disease are equally likely to benefit from CE. The aim of this study was to evaluate the yield of CE compared with other modalities in symptomatic patients with suspected or established CD using meta-analysis. We performed a recursive literature search of prospective studies comparing the yield of CE to other modalities in patients with suspected or established CD. Data on yield among various modalities were extracted, pooled, and analyzed. Incremental yield (IY) (yield of CE--yield of comparative modality) and 95% confidence intervals (95% CI) of CE over comparative modalities were calculated. Subanalyses of patients with a suspected initial presentation of CD and those with suspected recurrent disease were also performed. Nine studies (n = 250) compared the yield of CE with small bowel barium radiography for the diagnosis of CD. The yield for CE versus barium radiography for all patients was 63% and 23%, respectively (IY = 40%, p < 0.001, 95% CI = 28-51%). Four trials compared the yield of CE to colonoscopy with ileoscopy (n = 114). The yield for CE versus ileoscopy for all patients was 61% and 46%, respectively (IY = 15%, p= 0.02, 95% CI = 2-27%). Three studies compared the yield of CE to computed tomography (CT) enterography/CT enteroclysis (n = 93). The yield for CE versus CT for all patients was 69% and 30%, respectively (IY = 38%, p= 0.001, 95% CI = 15-60%). Two trials compared CE to push enteroscopy (IY = 38%, p < 0.001, 95% CI = 26-50%) and one trial compared CE to small bowel magnetic resonance imaging (MRI) (IY = 22%, p= 0.16, 95% CI =-9% to 53%). Subanalysis of patients with a suspected initial presentation of CD showed no statistically significant difference between the yield of CE and barium radiography (p= 0.09), colonoscopy with ileoscopy (p= 0.48), CT enterography (p= 0.07), or push enteroscopy (p= 0.51). Subanalysis of patients with established CD with suspected small bowel recurrence revealed a statistically significant difference in yield in favor of CE compared with all other modalities (barium radiography (p < 0.001), colonoscopy with ileoscopy (p= 0.002), CT enterography (p < 0.001), and push enteroscopy (p < 0.001)). In study populations, CE is superior to all other modalities for diagnosing non-stricturing small bowel CD, with a number needed to test (NNT) of 3 to yield one additional diagnosis of CD over small bowel barium radiography and NNT = 7 over colonoscopy with ileoscopy. These results are due to a highly significant IY with CE over all other modalities in patients with established non-stricturing CD being evaluated for a small bowel recurrence. While there was no significant difference seen between CE and alternate modalities for diagnosing small bowel CD in patients with a suspected initial presentation of CD, the trend toward significance for a number of modalities suggests the possibility of a type II error. Larger studies are needed to better establish the role of CE for diagnosing small bowel CD in patients with a suspected initial presentation of CD.


Simulated Saccadic Stimuli Suppress ON-Type Direction-Selective Retinal Ganglion Cells via Glycinergic Inhibition.
Two types of mammalian direction-selective ganglion cells (DSGCs), ON and ONOFF, operate over different speed ranges. The directional axes of the ON-DSGCs are thought to align with the axes of the vestibular system and provide sensitivity at rotational velocities that are too slow to activate the semicircular canals. ONOFF-DSGCs respond to faster image velocities. Using natural images that simulate the natural visual inputs to freely moving animals, we show that simulated visual saccades suppress responses in ON-DSGCs but not ONOFF-DSGCs recorded in retinas of domestic rabbits of either gender. Analysis of the synaptic inputs shows that this saccadic suppression results from glycinergic inputs that are specific to ON-DSGCs and are absent in ONOFF-DSGCs. When this glycinergic input is blocked, both cell types respond similarly to visual saccades and display essentially identical speed tuning. The results demonstrate that glycinergic circuits within the retina can produce saccadic suppression of retinal ganglion cell activity. The cell-type-specific targeting of the glycinergic circuits further supports the proposed physiological roles of ON-DSGCs in retinal-image stabilization and of ONOFF-DSGCs in detecting local object motion and signaling optical flow.SIGNIFICANCE STATEMENT In the mammalian retina, ON direction-selective ganglion cells (DSGCs) respond preferentially to slow image motion, whereas ONOFF-DSGCs respond better to rapid motion. The mechanisms producing this different speed tuning remain unclear. Here we show that simulated visual saccades suppress ON-DSGCs, but not ONOFF-DSGCs. This selective saccadic suppression is because of the selective targeting of glycinergic inhibitory synaptic inputs to ON-DSGCs. The different saccadic suppression in the two cell types points to different physiological roles, consistent with their projections to distinct areas within the brain. ON-DSGCs may be critical for providing the visual feedback signals that contribute to stabilizing the image on the retina, whereas ONOFF-DSGCs may be important for detecting the onset of saccades or for signaling optical flow.


Perceptual color map in macaque visual area V4.
Colors distinguishable with trichromatic vision can be defined by a 3D color space, such as red-green-blue or hue-saturation-lightness (HSL) space, but it remains unclear how the cortex represents colors along these dimensions. Using intrinsic optical imaging and electrophysiology, and systematically choosing color stimuli from HSL coordinates, we examined how perceptual colors are mapped in visual area V4 in behaving macaques. We show that any color activates 1-4 separate cortical patches within "globs," millimeter-sized color-preferring modules. Most patches belong to different hue or lightness clusters, in which sequential representations follow the color order in HSL space. Some patches overlap greatly with those of related colors, forming stacks, possibly representing invariable features, whereas few seem positioned irregularly. However, for any color, saturation increases the activity of all its patches. These results reveal how the color map in V4 is organized along the framework of the perceptual HSL space, whereupon different multipatch activity patterns represent different colors. We propose that such distributed and combinatorial representations may expand the encodable color space of small cortical maps and facilitate binding color information to other image features.


Reporting of methodological studies in health research: a protocol for the development of the MethodologIcal STudy reportIng Checklist (MISTIC).
Methodological studies (ie, studies that evaluate the design, conduct, analysis or reporting of other studies in health research) address various facets of health research including, for instance, data collection techniques, differences in approaches to analyses, reporting quality, adherence to guidelines or publication bias. As a result, methodological studies can help to identify knowledge gaps in the methodology of health research and strategies for improvement in research practices. Differences in methodological study names and a lack of reporting guidance contribute to lack of comparability across studies and difficulties in identifying relevant previous methodological studies. This paper outlines the methods we will use to develop an evidence-based tool-the MethodologIcal STudy reportIng Checklist-to harmonise naming conventions and improve the reporting of methodological studies. We will search for methodological studies in the Cumulative Index to Nursing and Allied Health Literature, Cochrane Library, Embase, MEDLINE, Web of Science, check reference lists and contact experts in the field. We will extract and summarise data on the study names, design and reporting features of the included methodological studies. Consensus on study terms and recommended reporting items will be achieved via video conference meetings with a panel of experts including researchers who have published methodological studies. The consensus study has been exempt from ethics review by the Hamilton Integrated Research Ethics Board. The results of the review and the reporting guideline will be disseminated in stakeholder meetings, conferences, peer-reviewed publications, in requests to journal editors (to endorse or make the guideline a requirement for authors), and on the Enhancing the QUAlity and Transparency Of health Research (EQUATOR) Network and reporting guideline websites. We have registered the development of the reporting guideline with the EQUATOR Network and publicly posted this project on the Open Science Framework (www.osf.io/9hgbq).


Robust determination of differential abundance in shotgun proteomics using nonparametric statistics.
Label-free shotgun mass spectrometry enables the detection of significant changes in protein abundance between different conditions. Due to often limited cohort sizes or replication, large ratios of potential protein markers to number of samples, as well as multiple null measurements pose important technical challenges to conventional parametric models. From a statistical perspective, a scenario similar to that of unlabeled proteomics is encountered in genomics when looking for differentially expressed genes. Still, the difficulty of detecting a large fraction of the true positives without a high false discovery rate is arguably greater in proteomics due to even smaller sample sizes and peptide-to-peptide variability in detectability. These constraints argue for nonparametric (or distribution-free) tests on normalized peptide values, thus minimizing the number of free parameters, as well as for measuring significance with permutation testing. We propose such a procedure with a class-based statistic, no parametric assumptions, and no parameters to select other than a nominal false discovery rate. Our method was tested on a new dataset which is available via ProteomeXchange with identifier PXD006447. The dataset was prepared using a standard proteolytic digest of a human protein mixture at 1.5-fold to 3-fold protein concentration changes and diluted into a constant background of yeast proteins. We demonstrate its superiority relative to other approaches in terms of the realized sensitivity and realized false discovery rates determined by ground truth, and recommend it for detecting differentially abundant proteins from MS data.


Reduced duration of breastfeeding is associated with a higher risk of multiple sclerosis in both Italian and Norwegian adult males: the EnvIMS study.
Breastfeeding for at least 4 months has been found to be associated with a reduced risk of immune-mediated diseases including multiple sclerosis (MS). Using data from a large multinational case-control study (EnvIMS), the association between MS and breastfeeding was investigated in two distinct populations. A questionnaire (EnvIMS-Q) which included a section on feeding during the first year of life was administered to MS cases and to age and sex frequency-matched controls from Italy and Norway. Logistic regression was used to estimate the odds ratio (ORs) and 95% confidence intervals (95% CIs) as a measure of the association between MS and exposure to prolonged breastfeeding (4 months or more, used as the reference category), vs. no breastfeeding or breastfeeding for less than 4 months (reduced exposure). Education, smoking habits, smoking in mother's pregnancy, and other types of milk used in infant feeding were included as covariates. A total of 547 cases and 1039 controls in Italy, and 737 cases and 1335 controls in Norway were studied. The distribution of prolonged (reference) breastfeeding differed between the Norwegian (65.4%) and the Italian (48.9%) study participants. A significant association between MS and reduced/no exposure to breastfeeding was found overall for Italy (OR(adj) = 1.37; 95% CI 1.09, 1.73), but not for Norway (OR(adj) = 1.14; 95% CI 0.92, 1.40). However, only in men, significant associations were observed for both populations (OR(Italy) = 2.33; 95% CI 1.50, 3.65, OR(Norway) = 2.13; 95% CI 1.37, 3.30). Reduced exposure to breastfeeding in males was found to be associated with increased risk of MS in Italy and in Norway.


Immune responses to influenza virus infection.
Influenza viruses cause annual outbreaks of respiratory tract infection with attack rates of 5-10%. This means that humans are infected repeatedly with intervals of, on average, 10-20 years. Upon each infection subjects develop innate and adaptive immune responses which aim at clearing the infection. Strain-specific antibody responses are induced, which exert selective pressure on circulating influenza viruses and which drive antigenic drift of seasonal influenza viruses, especially in the hemagglutinin molecule. This antigenic drift necessitates updating of seasonal influenza vaccines regularly in order to match the circulating strains. Upon infection also virus-specific T cell responses are induced, including CD4+ T helper cells and CD8+ cytotoxic T cells. These cells are mainly directed to conserved proteins and therefore display cross-reactivity with a variety of influenza A viruses of different subtypes. T cell mediated immunity therefore may contribute to so-called heterosubtypic immunity and may afford protection against antigenically distinct, potentially pandemic influenza viruses. At present, novel viral targets are identified that may help to develop broad-protective vaccines. Here we review the various arms of the immune response to influenza virus infections and their viral targets and discuss the possibility of developing universal vaccines. The development of such novel vaccines would imply that also new immune correlates of protection need to be established in order to facilitate assessment of vaccine efficacy.


Onset of puberty and near adult height in short children born small for gestational age and treated with GH: Interim analysis of up to 10 years of treatment in Japan.
The safety and effectiveness of long-term (10-yr) GH treatment in short Japanese children born small for gestational age (SGA) were evaluated based on interim data analysis from a clinical study, including the findings concerning the influence on the onset of puberty and subjects who achieved near adult height (NAH). Sixty-one subjects were analyzed at baseline in this study. Eleven subjects (6 boys and 5 girls) achieved NAH (mean 157.4 cm and 145.5 cm, respectively), and the Δ height SDS from the start of GH treatment was +1.6 in boys and +1.8 in girls. The median age (yr) at onset of puberty was 11.4 in boys and 9.9 in girls, comparable to healthy children. However, the mean height (cm) at onset of puberty (137.0 in boys; 125.5 in girls) was shorter than that of healthy children. Treatment-related adverse events were generally mild to moderate in severity; however, adenoidal hypertrophy was observed in two subjects as a serious adverse event. One subject had jaw malformation related to GH treatment at a dose of 0.067 mg/kg/d. No notable changes in HbA1c levels were observed, and the levels remained within the reference range. We have confirmed the safety and effectiveness of long-term GH treatment through this ongoing clinical study.


The mechanism of force generation in myosin: a disorder-to-order transition, coupled to internal structural changes.
We propose a molecular mechanism of force generation in muscle, based primarily on site-specific spectroscopic probe studies of myosin heads in contracting muscle fibers and myofibrils. Electron paramagnetic resonance (EPR) and time-resolved phosphorescence anisotropy (TPA) of probes attached to SH1 (Cys 707, in the catalytic domain of the head) have consistently shown that most myosin heads in contracting muscle are dynamically disordered, undergoing large-amplitude rotations in the microsecond time range. Some of these disordered heads are bound to actin, especially in the early (weak-binding, preforce) phase of the ATPase cycle. The small ordered population (10-20%) is rigidly oriented precisely as in rigor, with no other distinct angle observed in contraction or in the presence of intermediate states trapped by nucleotide analogs. These results are not consistent with the classical model in which the entire head undergoes a 45 degree transition between two distinct orientations. Therefore, it has been proposed that the catalytic domain of the myosin head has only one stereospecific (rigor-like) actin-binding angle, and that the head's internal structure changes during force generation, causing the distal light-chain-binding domain to rotate. To test this model, we have performed EPR and TPA studies of probes attached to regulatory light chains (RLCs) in rabbit and scallop myofibrils and fibers. The RLC results confirm the predominance of dynamic (microsecond) rotational disorder in both relaxation and contraction, and show that the different mechanisms of calcium regulation in the two muscles produce different rotational dynamics. In rabbit myofibrils, RLC probes are more dynamically disordered than SH1 probes, especially in rigor and contraction,indicating that the light-chain-binding domain undergoes rotational motions relative to the catalytic domain when myosin heads interact with actin. An SH1-bound spin label, which is sensitive to myosin's internal dynamics, resolves three distinct conformations during contraction, and time-resolved EPR shows that these transitions are coupled to specific steps in the ATPase cycle. We propose that force is generated during contraction by a disorder-to-order transition, in which myosin heads first attach weakly to actin in a nonstereospecific mode characterized by large-scale dynamic disorder, then undergo at least two conformational transitions involving large-scale structural (rotational) changes within the head, culminating in a highly ordered strong-binding state that bears force.


A population study comparing screening performance of prototypes for depression and anxiety with standard scales.
Screening instruments for mental disorders need to be short, engaging, and valid. Current screening instruments are usually questionnaire-based and may be opaque to the user. A prototype approach where individuals identify with a description of an individual with typical symptoms of depression, anxiety, social phobia or panic may be a shorter, faster and more acceptable method for screening. The aim of the study was to evaluate the accuracy of four new prototype screeners for predicting depression and anxiety disorders and to compare their performance with existing scales. Short and ultra-short prototypes were developed for Major Depressive Disorder (MDD), Generalised Anxiety Disorder (GAD), Panic Disorder (PD) and Social Phobia (SP). Prototypes were compared to typical short and ultra-short self-report screening scales, such as the Centre for Epidemiology Scale, CES-D and the GAD-7, and their short forms. The Mini International Neuropsychiatric Interview (MINI) version 6 1 was used as the gold standard for obtaining clinical criteria through a telephone interview. From a population sample, 225 individuals who endorsed a prototype and 101 who did not were administered the MINI. Receiver operating characteristic (ROC) curves were plotted for the short and ultra short prototypes and for the short and ultra short screening scales. The study found that the rates of endorsement of the prototypes were commensurate with prevalence estimates. The short-form and ultra short scales outperformed the short and ultra short prototypes for every disorder except GAD, where the GAD prototype outperformed the GAD 7. The findings suggest that people may be able to self-identify generalised anxiety more accurately than depression based on a description of a prototypical case. However, levels of identification were lower than expected. Considerable benefits from this method of screening may ensue if our prototypes can be improved for Major Depressive Disorder, Social Phobia and Panic Disorder.


The Blood Transcriptome of Experimental Melioidosis Reflects Disease Severity and Shows Considerable Similarity with the Human Disease.
Melioidosis, a severe human disease caused by the bacterium Burkholderia pseudomallei, has a wide spectrum of clinical manifestations ranging from acute septicemia to chronic localized illness or latent infection. Murine models have been widely used to study the pathogenesis of infection and to evaluate novel therapies or vaccines, but how faithfully they recapitulate the biology of human melioidosis at a molecular level is not known. In this study, mice were intranasally infected with either high or low doses of B. pseudomallei to generate either acute, chronic, or latent infection and host blood and tissue transcriptional profiles were generated. Acute infection was accompanied by a homogeneous signature associated with induction of multiple innate immune response pathways, such as IL-10, TREM1, and IFN signaling, largely found in both blood and tissue. The transcriptional profile in blood reflected the heterogeneity of chronic infection and quantitatively reflected the severity of disease. Genes associated with fibrosis and tissue remodeling, including matrix metalloproteases and collagen, were upregulated in chronically infected mice with severe disease. Transcriptional signatures of both acute and chronic melioidosis revealed upregulation of iNOS in tissue, consistent with the expression of IFN-γ, but also Arginase-1, a functional antagonist of the iNOS pathway, and was confirmed by immunohistochemistry. Comparison of these mouse blood datasets by pathway and modular analysis with the blood transcriptional signature of patients with melioidosis showed that many genes were similarly perturbed, including Arginase-1, IL-10, TREM1, and IFN signaling, revealing the common immune response occurring in both mice and humans.


'I never thought that this baby would survive; I thought that it would die any time': perceptions and care for preterm babies in eastern Uganda.
To explore the current care for and perceptions about preterm babies among community members in eastern Uganda. A neonatal midwife observed care of preterm babies in one general hospital and 15 health centres using a checklist and a field diary. In-depth interviews were conducted with 11 community health workers (CHWs) and also with 10 mothers, six fathers and three grandmothers of preterm babies. Three focus group discussions were conducted with midwives and women and men in the community. Content analysis of data was performed. Community members mentioned many features which may correctly be used to identify preterm babies. Care practices for preterm babies at health facilities and community level were inadequate and potentially harmful. Health facilities lacked capacity for care of preterm babies in terms of protocols, health workers' skills, basic equipment, drugs and other supplies. However, community members and CHWs stated that they accepted the introduction of preterm care practices such as skin-to-skin and kangaroo mother care. In this setting, care for preterm babies is inadequate at both health facility and community level. However, acceptance of the recommended newborn care practices indicated by the community is a window of opportunity for introducing programmes for preterm babies. In doing so, consideration needs to be given to the care provided at health facilities as well as to the gaps in community care that are largely influenced by beliefs, perceptions and lack of awareness.


A learned component of the ventilatory response to exercise in man.
The ventilatory response to mild-to-moderate exercise in humans is isocapnic, or 'error-free'. It has been suggested that this response is learned over many repetitions of exercise through the process of minimising any deviations from normal in the blood gas tensions, as sensed by the chemoreceptors. However, relatively limited training programmes have failed to produce any convincing evidence in humans that forcibly altering the blood gas tensions during repeated periods of exercise alters the subsequent steady-state ventilatory response to exercise. In this study, eight healthy young subjects were exposed, over a 7 day training period, to a total of 70 repeated bouts of exercise paired with a simultaneous airway CO2 load to stimulate the chemoreceptors (protocol EX + CO2). The ventilatory response to exercise was measured before and after training to determine whether it had been modified. Two further training protocols were undertaken as controls. One employed repeated exercise without an airway CO2 load, and the other employed repeated airway CO2 loading without exercise. On the 1st and 2nd days following training with protocol EX + CO2, end-tidal PCO2 was regulated at a lower level during steady-state exercise than following training with the control protocols and than before training (mean +/- S.E.M. reduction in end-tidal PCO2 = 1.32 +/- 0.36 Torr, ANOVA, P < 0.05). In contrast to previous studies, this finding demonstrates that the steady-state ventilatory response to exercise can be modified by a prior period of altered chemoreception during exercise. This suggests that ventilation is matched to metabolic rate during exercise by a mechanism that involves learning and memory.


Lef1DeltaN binds beta-catenin and increases osteoblast activity and trabecular bone mass.
Lymphoid enhancer-binding factor (Lef) 1 is a high mobility group protein best known as a Wnt-responsive transcription factor that associates with β-catenin. Lef1ΔN is a short isoform of Lef1 that lacks the first 113 amino acids and a well characterized high affinity β-catenin binding domain present in the full-length protein. Both Lef1 isoforms bind DNA and regulate gene expression. We previously reported that Lef1 is expressed in proliferating osteoblasts and blocks osteocalcin expression. In contrast, Lef1ΔN is only detectable in the later stages of osteoblast differentiation and promotes osteogenesis in vitro. Here, we show that Lef1ΔN retains the ability to interact physically and functionally with β-catenin. Unlike what has been reported in T cells and colon cancer cell lines, Lef1ΔN activated gene transcription in the absence of exogenous β-catenin and cooperated with constitutively active β-catenin to stimulate gene transcription in mesenchymal and osteoblastic cells. Residues at the N terminus of Lef1ΔN were required for β-catenin binding and the expression of osteoblast differentiation genes. To determine the role of Lef1ΔN on bone formation in vivo, a Lef1ΔN transgene was expressed in committed osteoblasts using the 2.3-kb fragment of the type 1 collagen promoter. The Lef1ΔN transgenic mice had higher trabecular bone volume in the proximal tibias and L5 vertebrae. Histological analyses of tibial sections revealed no differences in osteoblast, osteoid, or osteoclast surface areas. However, bone formation and mineral apposition rates as well as osteocalcin levels were increased in Lef1ΔN transgenic mice. Together, our data indicate that Lef1ΔN binds β-catenin, stimulates Lef/Tcf reporter activity, and promotes terminal osteoblast differentiation.


Experimentally comparing the attractiveness of domestic lights to insects: Do LEDs attract fewer insects than conventional light types?
LED lighting is predicted to constitute 70% of the outdoor and residential lighting markets by 2020. While the use of LEDs promotes energy and cost savings relative to traditional lighting technologies, little is known about the effects these broad-spectrum "white" lights will have on wildlife, human health, animal welfare, and disease transmission. We conducted field experiments to compare the relative attractiveness of four commercially available "domestic" lights, one traditional (tungsten filament) and three modern (compact fluorescent, "cool-white" LED and "warm-white" LED), to aerial insects, particularly Diptera. We found that LEDs attracted significantly fewer insects than other light sources, but found no significant difference in attraction between the "cool-" and "warm-white" LEDs. Fewer flies were attracted to LEDs than alternate light sources, including fewer Culicoides midges (Diptera: Ceratopogonidae). Use of LEDs has the potential to mitigate disturbances to wildlife and occurrences of insect-borne diseases relative to competing lighting technologies. However, we discuss the risks associated with broad-spectrum lighting and net increases in lighting resulting from reduced costs of LED technology.