Loss of transcriptional activation of the potassium channel Kir5.1 by HNF1β drives autosomal dominant tubulointerstitial kidney disease.
Hepatocyte nuclear factor 1 homeobox B (HNF1β) is an essential transcription factor for the development and functioning of the kidney. Mutations in HNF1β cause autosomal dominant tubulointerstitial kidney disease characterized by renal cysts and maturity-onset diabetes of the young (MODY). Moreover, these patients suffer from a severe electrolyte phenotype consisting of hypomagnesemia and hypokalemia. Until now, genes that are regulated by HNF1β are only partially known and do not fully explain the phenotype of the patients. Therefore, we performed chIP-seq in the immortalized mouse kidney cell line mpkDCT to identify HNF1β binding sites on a genome-wide scale. In total 7,421 HNF1β-binding sites were identified, including several genes involved in electrolyte transport and diabetes. A highly specific and conserved HNF1β site was identified in the promoter of Kcnj16 that encodes the potassium channel Kir5.1. Luciferase-promoter assays showed a 2.2-fold increase in Kcnj16 expression when HNF1β was present. Expression of the Hnf1β p.Lys156Glu mutant, previously identified in a patient with autosomal dominant tubulointerstitial kidney disease, did not activate Kcnj16 expression. Knockdown of Hnf1β in mpkDCT cells significantly reduced the appearance of Kcnj16 (Kir5.1) and Kcnj10 (Kir4.1) by 38% and 37%, respectively. These results were confirmed in a HNF1β renal knockout mouse which exhibited downregulation of Kcnj16, Kcnj10 and Slc12a3 transcripts in the kidney by 78%, 83% and 76%, respectively, compared to HNF1β wild-type mice. Thus, HNF1β is a transcriptional activator of Kcnj16. Hence, patients with HNF1β mutations may have reduced Kir5.1 activity in the kidney, resulting in hypokalemia and hypomagnesemia.


12(S)-hydroxyeicosatetraenoic acid and 13(S)-hydroxyoctadecadienoic acid regulation of protein kinase C-alpha in melanoma cells: role of receptor-mediated hydrolysis of inositol phospholipids.
Protein kinase C (PKC) isoenzymes are essential components of cell signaling. In this study, we investigated the regulation of PKC-alpha in murine B16 amelanotic melanoma (B16a) cells by the monohydroxy fatty acids 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and 13(S)-hydroxyoctadecadienoic acid [13(S)-HODE]. 12(S)-HETE induced a translocation of PKC-alpha to the plasma membrane and focal adhesion plaques, leading to enhanced adhesion of B16a cells to the matrix protein fibronectin. However, 13(S)-HODE inhibited these 12(S)-HETE effects on PKC-alpha. A receptor-mediated mechanism of action for 12(S)-HETE and 13(S)-HODE is supported by the following findings. First, 12(S)-HETE triggered a rapid increase in cellular levels of diacylglycerol and inositol trisphosphate in B16a cells. 13(S)-HODE blocked the 12(S)-HETE-induced bursts of both second messengers. Second, the 12(S)-HETE-increased adhesion of B16a cells to fibronectin was sensitive to inhibition by a phospholipase C inhibitor and pertussis toxin. Finally, a high-affinity binding site (Kd = 1 nM) for 12(S)-HETE was detected in B16a cells, and binding of 12(S)-HETE to B16a cells was effectively inhibited by 13(S)-HODE (IC50 = 4 nM). In summary, our data provide evidence that regulation of PKC-alpha by 12(S)-HETE and 13(S)-HODE may be through a guanine nucleotide-binding protein-linked receptor-mediated hydrolysis of inositol phospholipids.


The elephant brain in numbers.
What explains the superior cognitive abilities of the human brain compared to other, larger brains? Here we investigate the possibility that the human brain has a larger number of neurons than even larger brains by determining the cellular composition of the brain of the African elephant. We find that the African elephant brain, which is about three times larger than the human brain, contains 257 billion (10(9)) neurons, three times more than the average human brain; however, 97.5% of the neurons in the elephant brain (251 billion) are found in the cerebellum. This makes the elephant an outlier in regard to the number of cerebellar neurons compared to other mammals, which might be related to sensorimotor specializations. In contrast, the elephant cerebral cortex, which has twice the mass of the human cerebral cortex, holds only 5.6 billion neurons, about one third of the number of neurons found in the human cerebral cortex. This finding supports the hypothesis that the larger absolute number of neurons in the human cerebral cortex (but not in the whole brain) is correlated with the superior cognitive abilities of humans compared to elephants and other large-brained mammals.


Linking biomechanics to mobility and disability in people with knee osteoarthritis.
Knee osteoarthritis is the leading cause of chronic disability among community-dwelling adults. The factors resulting in this disability extend beyond pathology. Therefore, understanding disability requires assessment of both pathology and the day-to-day consequences of knee osteoarthritis. This review develops a novel osteoarthritis construct, "careful mobility," by linking qualitative and quantitative methods to study both patient experience and biomechanics.


Picomolar analysis of flavins in biological samples by dynamic pH junction-sweeping capillary electrophoresis with laser-induced fluorescence detection.
Sensitive capillary electrophoresis (CE) methods are required for emerging areas of biochemical research such as the metabolome. In this report, dynamic pH junction-sweeping CE with laser-induced fluorescence (LIF) detection is applied as a robust single method to analyze trace amounts of three flavin derivatives, riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD), from several types of samples including bacterial cell extracts, recombinant protein, and biological fluids. Submicromolar amounts of flavin coenzymes were measured directly from formic acid cell extracts of Bacillus subtilis. Significant differences in flavin concentration were measured in cell extracts derived from either glucose or malate as the carbon source in the culture media. Quantitative assessment of FAD and FMN content from selected flavoenzymes was demonstrated after heat denaturation to release noncovalently bound coenzymes and deproteinization. This method was also applied to the analysis of free flavins in pooled human plasma and urine without the need for laborious off-line sample preconcentration. Picomolar detectability of flavins by CE-LIF detection was realized with on-line preconcentration (up to 15% capillary length used for injection) by dynamic pH junction-sweeping, resulting in a limit of detection (S/N = 3) of about 4.0 pM for FAD and FMN. This represents over a 60-fold improvement in concentration sensitivity compared to those of previous techniques using conventional injections. The method was validated in terms of reproducibility, sensitivity, linearity, and specificity. Flavin analysis by dynamic pH junction-sweeping CE-LIF offers a simple, yet sensitive way to analyze trace levels of flavin metabolites from complex biological samples.


Insights into the P. y. yoelii hepatic stage transcriptome reveal complex transcriptional patterns.
During their complex life cycle, malaria parasites adopt morphologically, biochemically and immunologically distinct forms. The intra-hepatic form is the least known, yet of established value in the induction of sterile immunity and as a target for chemoprophylaxis. Using Plasmodium yoelii as a model we present here a novel approach to the elucidation of the transcriptome of this poorly studied stage. Sequences from Plasmodium were obtained in 388 of the 3533 inserts (11%) isolated from liver stages cDNA obtained from optimized cultures with high yields. These corresponded to a total of 88 putative P. yoelii genes. The majority of the transcribed genes identified, code for predicted proteins of as yet unknown function. The RT-PCR analysis carried out for 29 of these genes, confirmed expression at the hepatic stage and provided evidence for complex patterns of genes transcription in the distinct stages found in the mosquito and vertebrate host. The results demonstrate the efficacy of the approach that can now be applied to further detailed analysis of the hepatic stage transcriptome of Plasmodium.


Atrophic glossitis: Etiology, serum autoantibodies, anemia, hematinic deficiencies, hyperhomocysteinemia, and management.
Atrophic glossitis (AG) is characterized by the partial or complete absence of filiform papillae on the dorsal surface of the tongue. AG may reflect the significant deficiencies of some major nutrients including riboflavin, niacin, pyridoxine, vitamin B12, folic acid, iron, zinc, and vitamin E. Moreover, protein-calorie malnutrition, candidiasis, Helicobacter pylori colonization, xerostomia, and diabetes mellitus are also the etiologies of AG. Our previous study found the serum gastric parietal cell antibody (GPCA), thyroglobulin antibody (TGA), and thyroid microsomal antibody (TMA) positivities in 26.7%, 28.4%, and 29.8% of 1064 AG patients, respectively. We also found anemia, serum iron, vitamin B12, and folic acid deficiencies, and hyperhomocysteinemia in 19.0%, 16.9%, 5.3%, 2.3%, and 11.9% of 1064 AG patients, respectively. Moreover, GPCA-positive AG patients tended to have relatively higher frequencies of hemoglobin, iron, and vitamin B12 deficiencies and hyperhomocysteinemia than GPCA-negative AG patients. Supplementations with vitamin BC capsules plus corresponding deficient hematinics for those AG patients with hematinic deficiencies can achieve complete remission of oral symptoms and AG in some AG patients. Therefore, it is very important to examine the complete blood count, serum hematinic, homocysteine, and autoantibody levels in AG patients before we start to offer treatments for AG patients.


Dehumanization, retributive and restorative justice, and aggressive versus diplomatic intergroup conflict resolution strategies.
The desire for justice can escalate or facilitate resolution of intergroup conflicts. Two studies investigated retributive and restorative notions of justice as the mediating factor of the effect of perceived outgroup sentience-an aspect of (mechanistic) dehumanization referring to the emotional depth attributed to others-on intergroup conflict resolution. Study 1 showed that for Palestinians, who see themselves as victims, perceived sentience of Israelis decreased retributive but increased restorative notions of justice, which, ultimately, increased support for conflict resolution by negotiation rather than political violence. Study 2 partially replicated Study 1's findings with Jewish Israelis. The role of perceived sentience and its relationship to retributive and restorative notions of justice in protracted and nonprotracted conflicts and their resolution is discussed.


Clinical characteristics in a kindred with early-onset Alzheimer's disease and their linkage to a G-->T change at position 2149 of the amyloid precursor protein gene.
Patients from five generations of a family developed presenile Alzheimer's disease (AD) early in the fifth decade. Recent memory, information-processing speed, sequential tracking, and conceptual reasoning were the earliest cognitive functions affected. Language and visuoperceptual skills were largely spared early in the course of the disease. Later, there were progressive cognitive deficits and inability to perform the activities of daily living. Death occurred, on average, 6 years after onset. Three autopsies in affected members revealed cerebral amyloid deposits and neurofibrillary tangles. Clinical and pathologic features were typical for familial AD. Direct DNA sequencing revealed a G-->T change at position 2149 of the amyloid precursor protein (APP) gene that resulted in the substitution of phenylalanine for valine in the transmembrane domain of the mature protein. This mutation was present in DNA from all four examined affected individuals and linked to the disease with a lod score of 3.25, and was the most probable cause of AD in this family.


Distension of the gall bladder inhibits sphincter of Oddi motility in humans.
Studies in animals have suggested a neural reflex between the gall bladder and the sphincter of Oddi. The aim of this study was to investigate whether sphincter of Oddi motility is altered by distension of the gall bladder in humans. Sphincter of Oddi motility was recorded intraoperatively in 10 patients undergoing elective cholecystectomy for gall stones. The manometry was performed by a triple lumen constantly perfused catheter which was introduced through the cystic duct and positioned across the sphincter of Oddi to record sphincter basal pressure, wave amplitude, and frequency of contractions. In five patients a separate catheter was introduced into the gall bladder after ligation of the cystic duct. This catheter was used to distend the gall bladder. Sphincter of Oddi pressures were measured before, during, and after the distension. In a separate control group of patients (n = 5) basal sphincter of Oddi activity was recorded without distension of the gall bladder. Distension of the gall bladder decreased sphincter of Oddi basal pressure from (mean (SD] 22.8 (8.5) mmHg to 18.6 (6.5) mmHg (p = 0.01, paired t test) and frequency of sphincter of Oddi contractions decreased from 2.6 (1.6) to 1.1 (1.3) contractions/min (p = 0.003, paired t test). The results were significantly different from those of the control group (p less than 0.05, unpaired t test) during the same time period (four minutes). Pulse rate and blood pressure were not affected by the gall bladder distension. The results suggest a local reflex between the gall bladder and the sphincter of Oddi that might be important in the regulation of the pressure within the bile ducts and flow across the sphincter. This reflex is likely to be neurally mediated and injuries to it may be important in the aetiology of postcholecystectomy sphincter of Oddi dysfunction.


Serum carotenoids and coronary heart disease. The Lipid Research Clinics Coronary Primary Prevention Trial and Follow-up Study.
To examine the relationship between total serum carotenoid levels and the risk of subsequent coronary heart disease (CHD) events. New analysis of a cohort from the Lipid Research Clinics Coronary Primary Prevention Trial and Follow-up Study (LRC-CPPT). The LRC-CPPT was a multicenter placebo-controlled trial of cholestyramine resin and CHD with a follow-up period of 13 years. Serum carotenoids were measured at baseline. The placebo group of the LRC-CPPT, which consisted of 1899 men aged 40 to 59 years with type II-a hyperlipidemia and without known preexisting CHD, cancer, or other major illnesses. Nonfatal myocardial infarctions and deaths attributable to CHD ascertained from hospital records, autopsy reports, and death certificates and reviewed by a panel of cardiologists. After adjustment for known CHD risk factors including smoking, serum carotenoids were inversely related to CHD events. Men in the highest quartile of serum carotenoids had an adjusted relative risk (RR) of 0.64 (95% confidence interval [CI], 0.44 to 0.92) compared with the lowest quartile. For men who never smoked, this RR was 0.28 (95% CI, 0.11 to 0.73). The LRC-CPPT participants with higher serum carotenoid levels had a decreased risk of incident CHD. This finding was stronger among men who never smoked.


Impaired nitric oxide-mediated flow-induced dilation in arterioles of spontaneously hypertensive rats.
We tested the hypothesis that impairment of flow-dependent dilator mechanisms of skeletal muscle arterioles is one of the underlying reasons for the increased peripheral resistance in hypertension. Isolated, cannulated arterioles (approximately 55 microns) of gracilis muscle of 12-week-old spontaneously hypertensive (SH) and normotensive Wistar (NW) rats were investigated. At a constant perfusion pressure (80 mm Hg), the active diameters of NW and SH arterioles were 57.7 +/- 1.9 and 51.5 +/- 3.2 microns, whereas their passive diameters (Ca(2+)-free solution) were 113.6 +/- 2.9 and 101.7 +/- 2.9 microns, respectively. Flow-induced dilation was elicited by increases in flow of the perfusion solution from 0 to 25 microL/min in 5-microL/min steps. This response was significantly less in arterioles of SH compared with NW rats. For example, at 25-microL/min flow, the diameter of arterioles of SH rats was approximately 56% less (P < .05) than those of NW rats. Indomethacin, an inhibitor of prostaglandin synthesis, significantly attenuated the flow-diameter curve in both strains of rats. In contrast, N omega-nitro-L-arginine, a nitric oxide synthase inhibitor, significantly shifted the flow-diameter curve to the right in NW rats, but it did not affect the flow-diameter curve in SH rats. Thus, the present findings demonstrate that in gracilis muscle arterioles of normotensive rats in response to increases in flow (shear stress), prostaglandins and nitric oxide are co-released, resulting in a dilation. In early hypertension, however, there is a reduced arteriolar dilation to increases in flow that is due to the impairment of the nitric oxide-mediated portion of the flow-dependent arteriolar dilation.


Effects of topiramate and gabapentin on cognitive abilities in healthy volunteers.
To evaluate the cognitive effects of topiramate (TPM) and gabapentin (GBP). Forty healthy volunteers were randomized to a 12-week course of TPM, GBP, or placebo. Doses were gradually escalated over 10 weeks to a maximum of 400 mg/day of TPM or 3,600 mg/day of GBP or to the highest tolerated dose. Subjects were interviewed and examined biweekly. Cognitive testing was performed prior to initiating the drug and again 12 weeks later, at least 2 weeks after achieving plateau dosing. For each subject and cognitive measure, test-retest Z scores were calculated based on regression equations derived from 73 healthy volunteers. Group comparisons utilized the Wilcoxon test. There were significant TPM vs GBP and TPM vs placebo differences in test-retest Z scores for four of six target cognitive measures (Digit Symbol, Story Recall, Selective Reminding, Controlled Oral Word Association), always indicating worse retest performance for subjects receiving TPM. Overall, 12 of 24 cognitive measures were similarly affected. TPM effects were large, and several target measures averaged >2 SD of negative change. One measure was significantly affected by GBP. Topiramate (TPM) impaired cognitive test performance, whereas gabapentin had minimal effects. The effects of TPM were of sufficient magnitude potentially to affect daily and occupational function.


The human histone H3 complement anno 2011.
Histones are highly basic, relatively small proteins that complex with DNA to form higher order structures that underlie chromosome topology. Of the four core histones H2A, H2B, H3 and H4, it is H3 that is most heavily modified at the post-translational level. The human genome harbours 16 annotated bona fide histone H3 genes which code for four H3 protein variants. In 2010, two novel histone H3.3 protein variants were reported, carrying over twenty amino acid substitutions. Nevertheless, they appear to be incorporated into chromatin. Interestingly, these new H3 genes are located on human chromosome 5 in a repetitive region that harbours an additional five H3 pseudogenes, but no other core histone ORFs. In addition, a human-specific novel putative histone H3.3 variant located at 12p11.21 was reported in 2011. These developments raised the question as to how many more human histone H3 ORFs there may be. Using homology searches, we detected 41 histone H3 pseudogenes in the current human genome assembly. The large majority are derived from the H3.3 gene H3F3A, and three of those may code for yet more histone H3.3 protein variants. We also identified one extra intact H3.2-type variant ORF in the vicinity of the canonical HIST2 gene cluster at chromosome 1p21.2. RNA polymerase II occupancy data revealed heterogeneity in H3 gene expression in human cell lines. None of the novel H3 genes were significantly occupied by RNA polymerase II in the data sets at hand, however. We discuss the implications of these recent developments.


The role of genetic factors in the development of hyperhomocysteinemia.
Moderate hyperhomocysteinemia has been identified as a new independent risk factor for cardiovascular and neurodegenerative diseases. This fact has produced interest in the study of genetic variants involved in homocysteine metabolism and its relationship to pathogenesis. Recently, more than 15 different genes were studied for their relationship to plasma homocysteine levels. We determined the influence of genetic variants in five genes (5,10-methylenetetrahydrofolate reductase (MTHFR) 677C --> T, serine hydroxymethyltransferase (SHMT) 1420C --> T, thymidylate synthase (TS) 2R --> 3R, catechol-O-methyltransferase (COMT) 1947G --> A and transcobalamin (TC) 776C --> G) on plasma homocysteine, folic acid and parameters of vitamin B12 metabolism in 111 vegetarians (mean age: 46 +/- 15 years) and 118 healthy seniors (mean age: 82 +/- 6.5 years). Median homocysteine concentration in plasma was significantly influenced by the MTHFR genotypes in both populations. In the vegetarians the median homocysteine level was increased by 8 micromol/l in individuals homozygous for the mutation as compared to wild-type or heterozygous genotypes (20.4 micromol/l vs. 12.9 and 12.7 micromol/l, respectively). This unexpected increase was observed although the folate levels were in medium to elevated ranges. Our results suggest that vegetarians have a higher demand for folate to neutralize the genotype effect. Preclinical vitamin B12 deficiency in vegetarians may be the cause for disturbed remethylation and folate trap. Plasma homocysteine was not significantly influenced by the SHMT, TS, COMT and TC mutations. In addition, for the TC mutation a trend toward cellular vitamin B12 deficiency was observed. The methylmalonic acid (MMA) levels were slightly elevated and the holotranscobalamin-II (holoTC-II) levels decreased. In the vegetarian group a significant relationship between the COMT genotype and holoTC-II concentration in plasma was determined, whereas the high activity COMT genotype (G/G) resulted in increased levels (35 micromol/l vs. 21 micromol/l for heterozygous and low activity genotypes). The MMA levels were inversely correlated to holoTC-II concentrations. In conclusion, the study on vegetarians and seniors documents interesting lifestyle-genotype interactions. Although the TC and COMT mutations influence cellular vitamin B12 metabolism, this effect did not result in overt homocysteine elevation.


Detection of Vaccine-Induced Antibodies to Ebola Virus in Oral Fluid.
Blood sampling to assess production of antigen-specific antibodies after immunization is commonly performed, but it presents logistical difficulties for trials carried out during an infectious disease outbreak. In this study, we show that antibodies may be reliably detected in oral fluid collected in a minimally invasive manner without use of sharps. Clinical Trials Registration. NCT02240875.


TETT-functionalized TiO2 nanoparticles for DOX loading: a quantum mechanical study at the atomic scale.
In this work, we present a quantum mechanical investigation, based on the self-consistent charge density functional tight-binding (SCC-DFTB) method, of the functionalization with silane-type ligands (TETT) of a spherical TiO2 nanoparticle of realistic size (2.2 nm containing 700 atoms) to create an efficient nanosystem for simultaneous photodynamic therapy and drug transport. We determine the mechanism of the TETT ligand anchoring and its stability under thermal treatment, through molecular dynamics simulations at 300 K. Then, we build a medium and a full coverage model (22 and 40 TETTs, respectively) and analyze the interaction among TETT ligands and between the ligands and the surface. Finally, on the fully covered nanoparticle, we succeed in localizing two minimum energy structures for an attached doxorubicin anticancer molecule (DOX) and provide the atomistic details for both the covalent and the non-covalent (electrostatic) types of interaction. A future development of this work will be the investigation of the loading capacity of this drug delivery system and of the pH effect of the surrounding aqueous environment.


Women with impaired glucose tolerance during pregnancy have significantly poor pregnancy outcomes.
This article tests the hypothesis that women with impaired glucose tolerance (IGT) have the same pregnancy outcomes as those of their counterparts with normal glucose tolerance. From December 1998 to December 1999, 84 of 90 antenatal care base units (ACBUs) under the Tianjin Antenatal Care Network in China participated in the first screening program for gestational diabetes mellitus (GDM). A total of 9,471 pregnant women under the care of participating ACBUs were screened. Of the women screened, 154 were positive for IGT. Of the 154 women, 102 opted for conventional obstetric care. The comparison group was 302 women of normal glucose tolerance (NGT). The initial screening consisted of a 50-g 1-h glucose test, and was carried out at 26-30 gestational weeks. Women with a serum glucose > or =7.8 mmol/l were followed up with a 75-g 2-h oral glucose tolerance test. The World Health Organization's diagnostic criteria for GDM were used. Women with IGT were at increased risk for premature rupture of membranes (P-ROM) (odds ratio [OR] 10.07; 95% CI 2.90-34.93); preterm birth (6.42; 1.46-28.34); breech presentation (3.47; 1.11-10.84); and high birth weight (90th percentile or 4,000 g) (2.42; 1.07-5.46); adjusting for maternal age, pregravid BMI, hospital levels, and other confounding factors. The presence of IGT in pregnancy is predictive of poor pregnancy outcomes.


Putting cholesterol in its place: apoE and reverse cholesterol transport.
To avoid toxic overload of cholesterol in peripheral cells, the reverse cholesterol transport pathway directs excess cholesterol through HDL acceptors to the liver for elimination. In this issue of the JCI, a study by Matsuura et al. reveals new features of this pathway, including the importance of the ATP-binding cassette transporter G1 in macrophages and apoE in cholesteryl efflux from cells to cholesterol ester-rich (CE-rich) HDL(2) acceptors (see the related article beginning on page 1435). One proposal for boosting reverse cholesterol transport has been to elevate plasma HDL levels by inhibiting CE transfer protein (CETP), which transfers CE from HDL to lower-density lipoproteins. However, there has been concern that large, CE-rich HDL(2) generated by CETP inhibition might impair reverse cholesterol transport. ApoE uniquely facilitates reverse cholesterol transport by allowing CE-rich core expansion in HDL. In lower species, these large HDLs are not atherogenic. Thus, CETP might not be essential for reverse cholesterol transport in humans, raising hope of using a CETP inhibitor to elevate HDL levels.


The serum kinetics of bovine testicular hyaluronidase in dogs, rats and humans.
There is experimental and clinical evidence that i.v. injection of bovine testicular hyaluronidase (BTH) reduces the extent of necrosis during myocardial infarction. The fate of i.v. administered BTH has not been described. In this study, serum kinetics of BTH enzyme activity in dogs, rats and humans were determined. Tissue distribution of BTH was determined with an 125I-labeled preparation of purified BTH. Serum BTH activity initially decreased exponentially with half-life 2.0 +/- 0.1 min in dogs with coronary artery occlusion (n = 8; 500 U of BTH/kg); 3.2 min in humans with acute myocardial infarction (n = 2; 500 U of BTH/kg); and 3.2 +/- 0.3 min in rats (n = 5; 5,000 U of BTH/kg). In dogs BTH disappearance showed two distinct phases. After injection of high dose BTH (5,000 U of BTH/kg), during the first 7 min serum half-life of BTH was 2.1 +/- 0.2 min (n = 8), but increased to 9.4 min in later serum samples. After the injection of 125I-labeled BTH into the rat, protein-bound 125I disappeared from serum with a half-life (3.4 min) that is similar to the serum half-life of BTH enzyme activity (3.2 min). Twenty minutes after injection of 125I-labeled BTH, 30% of the label was recovered in the liver. It is concluded that BTH activity has a short serum half-life of less than 10 min in dogs, rats and humans. In the rat model, the disappearance of serum BTH activity results from physical removal of circulating BTH molecules rather than serum inhibition or inactivation of BTH enzymatic activity.


Contrasting the co-variability of daytime cloud and precipitation over tropical land and ocean.
The co-variability of cloud and precipitation in the extended tropics (35°N-35°S) is investigated using contemporaneous data sets for a 13-year period. The goal is to quantify potential relationships between cloud type fractions and precipitation events of particular strength. Particular attention is paid to whether the relationships exhibit different characteristics over tropical land and ocean. A primary analysis metric is the correlation coefficient between fractions of individual cloud types and frequencies within precipitation histogram bins that have been matched in time and space. The cloud type fractions are derived from Moderate Resolution Imaging Spectroradiometer (MODIS) joint histograms of cloud top pressure and cloud optical thickness in 1°grid cells, and the precipitation frequencies come from the Tropical Rainfall Measuring Mission (TRMM) Multi-satellite Precipitation Analysis (TMPA) data set aggregated to the same grid. It is found that the strongest coupling (positive correlation) between clouds and precipitation occurs over ocean for cumulonimbus clouds and the heaviest rainfall. While the same cloud type and rainfall bin are also best correlated over land compared to other combinations, the correlation magnitude is weaker than over ocean. The difference is attributed to the greater size of convective systems over ocean. It is also found that both over ocean and land the anti-correlation of strong precipitation with "weak" (i.e., thin and/or low) cloud types is of greater absolute strength than positive correlations between weak cloud types and weak precipitation. Cloud type co-occurrence relationships explain some of the cloud-precipitation anti-correlations. Weak correlations between weaker rainfall and clouds indicate poor predictability for precipitation when cloud types are known, and this is even more true over land than over ocean.


Monosomy of chromosome 3 and an inflammatory phenotype occur together in uveal melanoma.
In uveal melanoma, different predictors of poor prognosis have been identified, including monosomy of chromosome 3, HLA expression, and the presence of infiltrating leukocytes and macrophages. Each of these parameters can be used to differentiate prognostically the favorable tumors from the unfavorable ones, and thus the hypothesis for the present study was that they are related, and that monosomy of chromosome 3 occurs in the same tumors as the unfavorable inflammatory phenotype. Tumor tissue was obtained from 50 cases of uveal melanoma treated between 1999 and 2004. After enucleation, nuclei were isolated from paraffin-embedded tissue for fluorescence in situ hybridization, to determine the chromosome 3 copy number. Each tumor-containing globe was further processed for conventional histopathologic examination and for immunohistochemical analysis with HLA class I and II-specific antibodies and with macrophage marker CD68. Of 50 uveal melanomas, 62% (31/50) were categorized as having monosomy of chromosome 3. Monosomy 3 was associated with the presence of epithelioid cells, an increased density of tumor-infiltrating macrophages, and a higher HLA class I and II expression. Survival analyses showed a correlation between monosomy 3 and decreased survival and identified monosomy 3, ciliary body involvement, and largest basal tumor diameter as the best prognostic markers. Monosomy 3 in uveal melanoma is associated with the presence of an inflammatory phenotype, consisting of a high HLA class I and II expression as well as an increased number of tumor-infiltrating macrophages. In a multivariate Cox regression analysis, the presence of monosomy 3 was one of the best prognostic markers of metastatic disease and survival, although the follow-up time was short.


Acetaldehyde exchange above a managed temperate mountain grassland.
An overview of acetaldehyde exchange above a managed temperate mountain grassland in Austria over four growing seasons is presented. The meadow acted as a net source of acetaldehyde in all four years, emitting between 7 and 28 mg C m-2 over the whole growing period. The cutting of the meadow resulted in huge acetaldehyde emission bursts on the day of harvesting or one day later. During undisturbed conditions, both uptake and emission fluxes were recorded. The bidirectional nature of acetaldehyde fluxes was also reflected by clear diurnal cycles during certain time periods, indicating strong deposition processes before the 1st cut and emission towards the end of the growing season. The analysis of acetaldehyde compensation points revealed a complex relationship between ambient acetaldehyde mixing ratios and respective fluxes, significantly influenced by multiple environmental parameters and variable throughout the year. As a major finding of this study, we identified both a positive and negative correlation between concentration and flux on a daily scale, where soil temperature and soil water content were the most significant factors in determining the direction of the slope. In turn, this bidirectional relationship on a daily scale resulted in compensation points between 0.40 ppbv and 0.54 ppbv, which could be well explained by collected ancillary data. We conclude that in order to model acetaldehyde fluxes at the site in Neustift on a daily scale over longer time periods, it is crucial to know the type of relationship, i.e. the direction of the slope, between mixing ratios and fluxes on a given day.


Host cell responses to persistent mycoplasmas--different stages in infection of HeLa cells with Mycoplasma hominis.
Mycoplasma hominis is a facultative human pathogen primarily associated with bacterial vaginosis and pelvic inflammatory disease, but it is also able to spread to other sites, leading to arthritis or, in neonates, meningitis. With a minimal set of 537 annotated genes, M. hominis is the second smallest self-replicating mycoplasma and thus an ideal model organism for studying the effects of an infectious agent on its host more closely. M. hominis adherence, colonisation and invasion of HeLa cells were characterised in a time-course study using scanning electron microscopy, confocal microscopy and microarray-based analysis of the HeLa cell transcriptome. At 4 h post infection, cytoadherence of M. hominis to the HeLa cell surface was accompanied by differential regulation of 723 host genes (>2 fold change in expression). Genes associated with immune responses and signal transduction pathways were mainly affected and components involved in cell-cycle regulation, growth and death were highly upregulated. At 48 h post infection, when mycoplasma invasion started, 1588 host genes were differentially expressed and expression of genes for lysosome-specific proteins associated with bacterial lysis was detected. In a chronically infected HeLa cell line (2 weeks), the proportion of intracellular mycoplasmas reached a maximum of 10% and M. hominis-filled protrusions of the host cell membrane were seen by confocal microscopy, suggesting exocytotic dissemination. Of the 1972 regulated host genes, components of the ECM-receptor interaction pathway and phagosome-related integrins were markedly increased. The immune response was quite different to that at the beginning of infection, with a prominent induction of IL1B gene expression, affecting pathways of MAPK signalling, and genes connected with cytokine-cytokine interactions and apoptosis. These data show for the first time the complex, time-dependent reaction of the host directed at mycoplasmal clearance and the counter measures of this pestering pathogen.


Human neutrophils transform prostaglandins by a myeloperoxidase-dependent mechanism.
Intact human neutrophils, incubated with the soluble stimulant phorbol myristate acetate, discharge lysosomal components, generate oxygen metabolites, and transform exogenous 6-keto-prostaglandin F1 alpha, prostaglandin E2, and prostaglandin F2 alpha as assessed by thin layer radiochromatography. Neutrophils alone were incapable of transforming the prostaglandins. The addition of catalase or the myeloperoxidase inhibitor, azide, protected all three prostaglandins from the phorbol-stimulated neutrophils. Neither superoxide dismutase, heat-inactivated catalase, nor albumin had any inhibitory effect in this system. A model system consisting of glucose-glucose oxidase, as a source of H2O2, purified myeloperoxidase, and chloride was also able to transform the prostaglandins in an identical fashion. Neither glucose-glucose oxidase alone nor glucose-glucose oxidase and myeloperoxidase under chloride-free conditions were able to mediate this transformation. Thus, it appears that intact human neutrophils can transform prostaglandins by a mechanism dependent on H2O2, the lysosomal enzyme myeloperoxidase, and chloride. Given the importance of prostaglandins in regulating immune function, neutrophil-dependent prostaglandin transformation could play a novel role in modulating the inflammatory response.


Comparison of skin testing and three in vitro assays for specific IgE in the clinical evaluation of immediate hypersensitivity.
A new assay, Pharmacia CAP System (PCS), for allergen-specific IgE (sIgE) was evaluated in 198 new patients presenting with respiratory symptoms to an urban allergy practice. An experienced allergist examined each patient and clinically assessed sensitivity to timothy, short ragweed, Alternaria tenuis, cat, or D. farinae. Puncture and selected intracutaneous skin tests (ST) with these inhalant extracts were then performed. The physician again rated the likelihood of clinical sensitivity to each inhalant, and serum was obtained for sIgE measurements by Phadebas RAST, modified RAST, and PCS. Results of the three in vitro tests (IVT) correlated well with each other and generally agreed with physician assessments and ST results. Individual differences for extracts and assay methods were identified. A few patients with negative ST had positive IVT, but more patients with positive ST were negative by IVT. Modified RAST had greater sensitivity but less specificity than the other two IVT. Analysis of receiver operating characteristic curves showed that sensitivity of the three assays when compared at the 95% level of specificity, did not differ. This result suggests that the cutoff criterion for a positive modified RAST result is too low and should be reevaluated. Skin tests remain the most sensitive and specific test available. The Pharmacia CAP System is a clinically useful assay for sIgE and appears to be a clear advancement for IVT technology.


The dimensions of the posterior arch of C2 for instrumented screw fixation. A radiological study in the Spanish population.
To describe the feasibility of the posterior arch of C2 accepting two crossing screws in the Spanish population. One hundred and fifty patients who underwent a routine neck CT scan for non-cervical pathology were enrolled. Submillimeter slices (thickness 0.7 mm) every 0.4 mm were performed with a 64 multi-detector CT scan, which allows isometric measurements. We measured the length and height of the cortical and cancellous (endomedullar) region of the lamina and the spinous process, inclination, maximal screw length and spinolaminar angle. The average (standard deviation) measurements of the lamina were: width of the left cortical 7.2 (1.5) mm, right cortical 6.9 (1.3) mm, width of the cancellous part of the left lamina 4.8 (1.5) mm, right side 4.6 (1.4) mm. The mean left cortical height was 13.0 (1.5) mm and 13.1 (1.6) mm for the right. The mean height of the cancellous part was 9.0 mm for both sides. The average measurements of the spinous process were: cortical length 15.7 (3.5) mm, endomedullar length 12.5 (3.9) mm; cortical height 11.9 (2.2) mm, endomedullar height 8.4 (2.1) mm; spinolaminar angle 49º (4); the maximum screw length 3.18 cm, and the inclination angle 143º. A CT scan with submillimeter slices is necessary in order to avoid malpositioning of the screws. The outer cortical measurements are 2 to 4 mm bigger than the endomedullar ones. Taking into account the dimensions of the spinous process, 24% of the population would not be candidates for this crossing screw technique.


Theiler's virus infection of beta 2-microglobulin-deficient mice.
Theiler's virus, a murine picornavirus, persists in the central nervous systems of susceptible mice and induces a chronic demyelinating disease. Susceptibility or resistance to this disease is controlled in part by the H2-D locus of the major histocompatibility complex (MHC). For this reason, it has been proposed that CD8+ class I-restricted cytotoxic T cells play a main role in the pathogenesis of this viral infection. We recently reported the existence of anti-virus CD8+ cytotoxic T cells in the course of Theiler's virus infection. In the present study, we examined the role of these effector cells in mice in which the beta 2-microglobulin gene had been disrupted. These mice fail to express class I MHC molecules and therefore lack CD8+ T cells. The mice are derived from a C57BL/6 x 129/Ola cross and are H-2b, a haplotype associated with resistance to Theiler's virus infection. beta 2-Microglobulin-deficient mice (beta 2m-/-mice) failed to clear the virus, developed demyelination, and, interestingly, did not succumb to early infection. These results demonstrate that CD8+ T cells are required to clear Theiler's virus infection. In contrast with a current hypothesis, they also demonstrate that CD8+ T cells are not major mediators of the demyelinating disease.


Quantitative podocyte parameters predict human native kidney and allograft half-lives.
Kidney function decreases with age. A potential mechanistic explanation for kidney and allograft half-life has evolved through the realization that linear reduction in glomerular podocyte density could drive progressive glomerulosclerosis to impact both native kidney and allograft half-lives. Predictions from podometrics (quantitation of podocyte parameters) were tested using independent pathologic, functional, and outcome data for native kidneys and allografts derived from published reports and large registries. With age, native kidneys exponentially develop glomerulosclerosis, reduced renal function, and end-stage kidney disease, projecting a finite average kidney life span. The slope of allograft failure rate versus age parallels that of reduction in podocyte density versus age. Quantitative modeling projects allograft half-life at any donor age, and rate of podocyte detachment parallels the observed allograft loss rate. Native kidneys are designed to have a limited average life span of about 100-140 years. Allografts undergo an accelerated aging-like process that accounts for their unexpectedly short half-life (about 15 years), the observation that older donor age is associated with shorter allograft half-life, and the fact that long-term allograft survival has not substantially improved. Podometrics provides potential readouts for these processes, thereby offering new approaches for monitoring and intervention. National Institutes of Health.


Juvenile hormone regulates butterfly larval pattern switches.
Insect color patterns can be very diverse. This variation is also seen among many larval instar stages, which can take on vastly different phenotypes. Young caterpillars of the swallowtail butterfly, Papilio xuthus, are mimics of bird droppings, whereas the fifth larval instar is camouflaged among the leaves of host plants (cryptic pattern). We find that juvenile hormone (JH) titers decrease during the fourth larval instar. Furthermore, treatment with JH analog at the beginning of the fourth instar stage resulted in reproducing the mimetic pattern instead of the usual cryptic one and likewise altered gene expression patterns to that associated with the mimetic pattern. These findings suggest that JH regulates the progressive larval pattern switch of this insect.


FRED--a framework for T-cell epitope detection.
Over the last decade, immunoinformatics has made significant progress. Computational approaches, in particular the prediction of T-cell epitopes using machine learning methods, are at the core of modern vaccine design. Large-scale analyses and the integration or comparison of different methods become increasingly important. We have developed FRED, an extendable, open source software framework for key tasks in immunoinformatics. In this, its first version, FRED offers easily accessible prediction methods for MHC binding and antigen processing as well as general infrastructure for the handling of antigen sequence data and epitopes. FRED is implemented in Python in a modular way and allows the integration of external methods. FRED is freely available for download at http://www-bs.informatik.uni-tuebingen.de/Software/FRED.


Repair of the secretion defect in the Z form of alpha 1-antitrypsin by addition of a second mutation.
Homozygous inheritance of the Z-type mutant form of the alpha 1-antitrypsin (alpha 1AT) gene results in the most common form of alpha 1AT deficiency, a human hereditary disease associated with a high risk for the development of emphysema and an increased incidence of neonatal hepatitis. The alpha 1AT-synthesizing cells of individuals with the Z gene have normal alpha 1AT messenger RNA levels, but alpha 1AT secretion is markedly reduced secondary to accumulation of newly synthesized alpha 1AT in the rough endoplasmic reticulum. Crystallographic analysis of alpha 1AT predicts that in normal alpha 1AT, a negatively charged Glu342 is adjacent to positively charged Lys290. Thus the Glu342----Lys342 Z mutation caused the loss of a normal salt bridge, resulting in the intracellular aggregation of the Z molecule. The prediction was made that a second mutation in the alpha 1AT genet that changed the positively charged Lys290 to a negatively charged Glu290 would correct the secretion defect. When the second mutation was added to the Z-type complementary DNA, the resulting gene directed the synthesis and secretion of amounts of alpha 1AT similar to that directed by the normal alpha 1AT complementary DNA in an in vitro eukaryotic expression system. This suggests the possibility that a human hereditary disease can be corrected by inserting an additional mutation in the same gene.


Maize RNA PolIV affects the expression of genes with nearby TE insertions and has a genome-wide repressive impact on transcription.
RNA-directed DNA methylation (RdDM) is a plant-specific epigenetic process that relies on the RNA polymerase IV (Pol IV) for the production of 24 nucleotide small interfering RNAs (siRNA) that guide the cytosine methylation and silencing of genes and transposons. Zea mays RPD1/RMR6 gene encodes the largest subunit of Pol IV and is required for normal plant development, paramutation, transcriptional repression of certain transposable elements (TEs) and transcriptional regulation of specific alleles. In this study we applied a total RNA-Seq approach to compare the B73 and rpd1/rmr6 leaf transcriptomes. Although previous studies indicated that loss of siRNAs production in RdDM mutants provokes a strong loss of CHH DNA methylation but not massive gene or TEs transcriptional activation in both Arabidopsis and maize, our total RNA-Seq analysis of rpd1/rmr6 transcriptome reveals that loss of Pol IV activity causes a global increase in the transcribed fraction of the maize genome. Our results point to the genes with nearby TE insertions as being the most strongly affected by Pol IV-mediated gene silencing. TEs modulation of nearby gene expression is linked to alternative methylation profiles on gene flanking regions, and these profiles are strictly dependent on specific characteristics of the TE member inserted. Although Pol IV is essential for the biogenesis of siRNAs, the genes with associated siRNA loci are less affected by the pol IV mutation. This deep and integrated analysis of gene expression, TEs distribution, smallRNA targeting and DNA methylation levels, reveals that loss of Pol IV activity globally affects genome regulation, pointing at TEs as modulator of nearby gene expression and indicating the existence of multiple level epigenetic silencing mechanisms. Our results also suggest a predominant role of the Pol IV-mediated RdDM pathway in genome dominance regulation, and subgenome stability and evolution in maize.


Challenges faced by rural people in mitigating the effects of climate change in the Mazungunye communal lands, Zimbabwe.
The phenomenon of climate change is one of the most contested and debated concepts globally. Some governments still deny the existence of climate change and its impact on rural-urban areas around the world. However, the effects of climate change have been visible in rural Zimbabwe, with some communities facing food insecurity, water scarcity and loss of livestock. Climate change has impacted negatively on agriculture, which is the main source of livelihood in Zimbabwe's rural communities. This study aims at exploring challenges faced by rural people in mitigating the effects of climate change in the Mazungunye community, Masvingo Province, in Zimbabwe. The objectives of the study were to identify the challenges that impede effective adaptation of rural people to climate change hazards and to examine their perceptions on how to foster effective adaptation. The researchers conducted a qualitative research study guided by descriptive and exploratory research designs. Purposive sampling was employed to draw the population of the study. The population sample consisted of 26 research participants drawn from members of the community. Data was collected through in-depth individual interviews and focus group discussions. Thematic content analysis was used to analyse data. The findings of the study revealed the following challenges: unpredictability of indigenous knowledge systems, lack of resources and technoscience adaptive methods, lack of support to implement viable mitigation strategies, lack of information about resilience and adaptive capacity to climate change. This study has significance to policymakers and other stakeholders concerned with devising and implementing policies and programmes that are responsive to rural people's needs in the climate change terrain, tapping into their presenting challenges as a departure point for intervention. The study recommended that the most important way to help rural poor people adapt to climate change is through the provision of information; immediate response to needs and climate-smart agricultural policies.


Efficacy and safety of intraoperative intracameral mydriasis in manual small incision cataract surgery - A randomized controlled trial.
The purpose of this study is to assess the efficacy and safety of intracameral mydriatic solution, as compared to preoperative topical mydriatics, in patients undergoing manual small incision cataract surgery (MSICS) under peribulbar anesthesia. To assess the sustainability of intracameral mydriasis in MSICS by monitoring pupil size at specific junctures during the surgery. This trial recruited 127 patients, who underwent MSICS under peribulbar block. Mydriasis in topical group was achieved with preoperative topical dilating drops while patients in intracameral group were taken up for surgery without dilation, and mydriasis was achieved intraoperatively with intracameral solution. Pupil sizes were measured serially, at six different junctures during surgery. Time duration of surgery, any intraoperative complications and first postoperative day visual acuity, corneal edema score, and anterior chamber inflammation score were noted in all patients. Mean pupil size just before peribulbar block was 7.3 mm in topical group and 3.3 mm in intracameral group (P < 0.001). Mean pupil size in intracameral group increased to 7.3 mm 30 s after injecting intracameral dilating solution. Mean pupil size in both groups progressively reduced, reaching 5.5 mm (topical group) and 6.2 mm (intracameral group) just before intraocular lens implantation (P = 0.001), and measured 5.1 mm and 5.5 mm, respectively, at the end of surgery (P = 0.048). On first postoperative day, there was no significant difference in distribution of corneal edema scores, AC inflammation scores, and in median logMAR visual acuity between the two groups. MSICS can be performed effectively and safely utilizing intracameral mydriatic solution, without the use of preoperative dilating drops. CTRI/2016/06/007036.


Prevalence-based economic evaluation.
Researchers have often stated that economic evaluations of new drugs have rarely been used to inform healthcare decisions, despite the large volume of published studies. In this paper, a new categorization for economic evaluations of new drugs is proposed: incidence-based and prevalence-based. This categorization is designed to increase the likelihood that decision-makers are given more complete and usable economic information about new treatments. Incidence-based evaluations (such as cost-effectiveness analysis) focus on the impact of a new treatment on a health condition from onset until cure or death. Prevalence-based evaluations focus on the impact of a new treatment on a health condition during a 1-year period. An incidence-based evaluation may focus either on a representative individual or on a specific disease cohort. A prevalence-based evaluation generally focuses on a specific population. Incidence-based evaluations measure the value of the new treatment compared to alternative treatments for the same health conditions and compared to commonly used treatments for other health conditions. Prevalence-based evaluations measure the impact of introducing the new treatment on annual healthcare budgets and population health. Both types of evaluation provide important information when a new treatment is introduced to a population.


Divergent evolutionary processes associated with colonization of offshore islands.
Oceanic islands have been a test ground for evolutionary theory, but here, we focus on the possibilities for evolutionary study created by offshore islands. These can be colonized through various means and by a wide range of species, including those with low dispersal capabilities. We use morphology, modern and ancient sequences of cytochrome b (cytb) and microsatellite genotypes to examine colonization history and evolutionary change associated with occupation of the Orkney archipelago by the common vole (Microtus arvalis), a species found in continental Europe but not in Britain. Among possible colonization scenarios, our results are most consistent with human introduction at least 5100 bp (confirmed by radiocarbon dating). We used approximate Bayesian computation of population history to infer the coast of Belgium as the possible source and estimated the evolutionary timescale using a Bayesian coalescent approach. We showed substantial morphological divergence of the island populations, including a size increase presumably driven by selection and reduced microsatellite variation likely reflecting founder events and genetic drift. More surprisingly, our results suggest that a recent and widespread cytb replacement event in the continental source area purged cytb variation there, whereas the ancestral diversity is largely retained in the colonized islands as a genetic 'ark'. The replacement event in the continental M. arvalis was probably triggered by anthropogenic causes (land-use change). Our studies illustrate that small offshore islands can act as field laboratories for studying various evolutionary processes over relatively short timescales, informing about the mainland source area as well as the island.


Temperature dependence of myofilament Ca sensitivity of rat, guinea pig, and frog ventricular muscle.
Cooling the superfusate of intact ventricular muscle, from 30 degrees C to below 4 degrees C in less than 2 s, leads to contractures thought to reflect the amount of Ca available for release from the sarcoplasmic reticulum (SR). On rewarming, tension transiently increases in guinea pig and rat ventricular muscle. It has been proposed that this rewarming tension spike reflects changes in myofilament Ca sensitivity and maximum Ca-activated force (Cmax) associated with rewarming. There are differences in intracellular Ca regulation among cardiac muscle preparations. Some characteristics of rapid-cooling contractures (e.g., the magnitude of the rewarming spike) also differ between species. Therefore, the Ca sensitivity of skinned ventricular muscle from the rat, guinea pig, and frog was determined at 29 (22 degrees C for frog ventricular preparations), 8, and 1 degrees C. The results show that cooling rat and guinea pig ventricular muscle from 29 to 1 degrees C shifts the pCa vs. tension relationship toward higher [Ca2+] by 0.65 and 0.55 pCa units, respectively. Cooling to 1 degrees C also reduced Cmax to 3.3 and 7.8% of that at 29 degrees C in rat and guinea pig ventricular muscle, respectively. Similar results were found for frog ventricular muscle, in which cooling from 22 to 1 degrees C reduced Ca sensitivity by 0.6 pCa units and Cmax to 45.7% of its value at 22 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical features of facioscapulohumeral muscular dystrophy 2.
In some 5% of patients with facioscapulohumeral muscular dystrophy (FSHD), no D4Z4 repeat contraction on chromosome 4q35 is observed. Such patients, termed patients with FSHD2, show loss of DNA methylation and heterochromatin markers at the D4Z4 repeat that are similar to patients with D4Z4 contractions (FSHD1). This commonality suggests that a change in D4Z4 chromatin structure unifies FSHD1 and FSHD2. The aim of our study was to critically evaluate the clinical features in patients with FSHD2 in order to establish whether these patients are phenotypically identical to FSHD1 and to establish the effects of the (epi-) genotype on the phenotype. This cross-sectional study studied 33 patients with FSHD2 from 27 families, the largest cohort described to date. All patients were clinically assessed using a standardized clinical evaluation form. Genotype analysis was performed by pulsed field gel electrophoresis and PCR; D4Z4 methylation was studied by methylation-sensitive Southern blot analysis. FSHD2 is identical to FSHD1 in its clinical presentation. Notable differences include a higher incidence (67%) of sporadic cases and the absence of gender differences in disease severity in FSHD2. Overall, average disease severity in FSHD2 was similar to that reported in FSHD1 and was not influenced by D4Z4 repeat size. In FSHD2, a small effect of the degree of hypomethylation on disease severity was observed. Clinically, patients with FSHD2 are indistinguishable from patients with FSHD1. The present data suggest that FSHD1 and FSHD2 are the result of the same pathophysiologic process.


The significance of droughts for hyporheic dwellers: evidence from freshwater crayfish.
Freshwater biodiversity is globally threatened by various factors while severe weather events like long-term droughts may be substantially devastating. In order to remain in contact with the water or stay in a sufficiently humid environment at drying localities, the ability to withstand desiccation by dwelling in the hyporheic zone, particularly through vertical burrowing is crucial. We assessed the ability of three European native and five non-native crayfish as models to survive and construct vertical burrows in a humid sandy-clayey substrate under a simulated one-week drought. Three native species (Astacus astacus, A. leptodactylus, and Austropotamobius torrentium) suffered extensive mortalities. Survival of non-native species was substantially higher while all specimens of Cherax destructor and Procambarus clarkii survived. The native species and Pacifastacus leniusculus exhibited no ability to construct vertical burrows. Procambarus fallax f. virginalis and P. clarkii constructed bigger and deeper burrows than C. destructor and Orconectes limosus. In the context of predicted weather fluctuations, the ability to withstand desiccation through constructing vertical burrows into the hyporheic zone under drought conditions might play a significant role in the success of particular crayfish species, as well as a wide range of further hyporheic-dwelling aquatic organisms in general.


Parents are a drag: long-lived birds share the cost of increased foraging effort with their offspring, but males pass on more of the costs than females.
Life history theory predicts that parents will balance benefits from investment in current offspring against benefits from future reproductive investments. Long-lived organisms are therefore less likely to increase parental effort when environmental conditions deteriorate. To investigate the effect of decreased foraging capacity on parental behaviour of long-lived monogamous seabirds, we experimentally increased energy costs for chick-rearing thick-billed murres (Uria lomvia). Handicapped birds had lighter chicks and lower provisioning rates, supporting the prediction that long-lived animals would pass some of the costs of impaired foraging ability on to their offspring. Nonetheless, handicapped birds spent less time underwater, had longer inter-dive surface intervals, had lower body mass, showed lower resighting probabilities in subsequent years and consumed fewer risky prey items. Corticosterone levels were similar between control and handicapped birds. Apparently, adults shared some of the costs of impaired foraging, but those costs were not measurable in all metrics. Handicapped males had higher plasma neutral lipid concentrations (higher energy mobilisation) and their chicks exhibited lower growth rates than handicapped females, suggesting different sex-specific investment strategies. Unlike other studies of auks, partners did not compensate for handicapping, despite good foraging conditions for unhandicapped birds. In conclusion, parental murres and their offspring shared the costs of experimentally increased foraging constraints, with females investing more than males.


Vortex generation in pulsatile flow through arterial bifurcation models including the human carotid artery.
Visualization experiments were performed to elucidate the complicated flow pattern in pulsatile flow through arterial bifurcations. Human common carotid arteries, which were made transparent, and glass-models simulating Y- and T-shaped bifurcations were used. Pulsatile flow with wave forms similar to those of arterial flow was generated with a piston pump, elastic tube, airchamber, and valves controlling the outflow resistance. Helically recirculating flow with a pattern similar to that of the horseshoe vortex produced around wall-based protuberances in circular tubes was observed in pulsatile flow through all the bifurcations used in the present study. This flow type, which we shall refer to as the horseshoe vortex, has also been demonstrated to occur at the human common carotid bifurcation in steady flow with Reynolds numbers above 100. Time-varying flows also produced the horseshoe vortex mostly during the decelerating phase. Fluid particles of dye solution approaching the bifurcation apex diverged, divided into two directions perpendicularly, and then showed helical motion representing the horseshoe vortex formation. While this helical flow was produced, the stagnation points appeared on the wall upstream of the apex. Their position was dependent upon the flow distribution ratio between the branches in the individual arteries. The region affected by the horseshoe vortex was smaller during pulsatile flow than during steady flow. Lowering the Reynolds number together with the Womersley number weakened the intensity of helical flow. A separation bubble, resulting from the divergence or wall roughness, was observed at the outer or inner wall of the branch vessels and made the flow more complicated.


Lack of correlation between serum anti-HBcore detectability and hepatocellular carcinoma in patients with HCV-related cirrhosis.
While the likelihood of developing hepatocellular carcinoma (HCC) in patients coinfected with both HBV and HCV is increased, the role of previous exposure to HBV as a risk factor associated with tumor occurrence in subjects with HCV-related cirrhosis has not been fully investigated. To assess whether serum anti-HBc positivity, as a marker of previous HBV exposure, is associated with HCC development in HCV-related positive, hepatitis B surface antigen (HBsAg) negative patients with cirrhosis treated with alfa-interferon (IFN) monotherapy. PATIENTS AND: A database including 883 consecutive patients (557 men, mean age 54.7 yr) with histologically proven cirrhosis treated with IFN between 1992 and 1997 was analyzed. All subjects have been surveilled every 6 months by ultrasound. Independent predictors of HCC were assessed by Cox multiple regression analysis. Mean follow-up was 96.1 months. Anti-HBc testing was available in 693 cases and, among them, 303 patients (43.7%) were anti-HBc seropositive. Anti-HBc positive patients were more often men (67.0%vs 58.7%, P= 0.03), had lower transaminase levels (3.3 +/- 2.0 vs 3.8 +/- 2.5 u.l.n., P= 0.004), and had higher rate of alcohol intake (38.3%vs 22.5%, P < 0.001) than anti-HBc negative patients. Overall, the incidence rates of HCC per 100 person-years were 1.84 (95% CI 1.34-2.47) in the anti-HBc positive patients and 1.86 (95% CI 1.41-2.42) in anti-HBc negative ones. By Cox multiple regression, there was no association of serum anti-HBc with HCC development (HR 1.03, 95% CI 0.69-1.52) or liver-related deaths incidence (HR 1.21; 95% CI 0.76-1.95). In comparison with anti-HBc negative subjects, serum anti-HBc positive patients with HCV-related/HBsAg negative cirrhosis treated with IFN monotherapy did not show a greater risk of HCC.


Introduction to a New Special Series for the Journal of Pain and Symptom Management-Science in Action: Evidence and Opportunities for Palliative Care Across Diverse Populations and Care Settings.
Practices to optimize palliative care delivery and new opportunities in which to integrate palliative care vary across populations and care settings. Systematic reviews are an efficient and methodologically rigorous approach to summarize existing research to identify both evidence-based best practices and new areas for future research and clinical practice. This is the introduction to a special series of articles in which members of the American Academy of Hospice and Palliative Medicine Research Committee report the results of circumscribed systematic reviews, which in a specific population or care setting seek to 1) summarize existing evidence for optimal palliative care practices or 2) identify opportunities where better palliative care delivery could improve patient and/or family outcomes.


Multi-genome identification and characterization of chlamydiae-specific type III secretion substrates: the Inc proteins.
Chlamydiae are obligate intracellular bacteria that multiply in a vacuolar compartment, the inclusion. Several chlamydial proteins containing a bilobal hydrophobic domain are translocated by a type III secretion (TTS) mechanism into the inclusion membrane. They form the family of Inc proteins, which is specific to this phylum. Based on their localization, Inc proteins likely play important roles in the interactions between the microbe and the host. In this paper we sought to identify and analyze, using bioinformatics tools, all putative Inc proteins in published chlamydial genomes, including an environmental species. Inc proteins contain at least one bilobal hydrophobic domain made of two transmembrane helices separated by a loop of less than 30 amino acids. Using bioinformatics tools we identified 537 putative Inc proteins across seven chlamydial proteomes. The amino-terminal segment of the putative Inc proteins was recognized as a functional TTS signal in 90% of the C. trachomatis and C. pneumoniae sequences tested, validating the data obtained in silico. We identified a macro domain in several putative Inc proteins, and observed that Inc proteins are enriched in segments predicted to form coiled coils. A surprisingly large proportion of the putative Inc proteins are not constitutively translocated to the inclusion membrane in culture conditions. The Inc proteins represent 7 to 10% of each proteome and show a great degree of sequence diversity between species. The abundance of segments with a high probability for coiled coil conformation in Inc proteins support the hypothesis that they interact with host proteins. While the large majority of Inc proteins possess a functional TTS signal, less than half may be constitutively translocated to the inclusion surface in some species. This suggests the novel finding that translocation of Inc proteins may be regulated by as-yet undetermined mechanisms.


Fluorinated derivatives of tetrahydroaltersolanol molecule on COVID-19, HIV, and HTLV protease by DFT and molecular docking approaches.
Structural, optoelectronic, and biological properties of tetrahydroaltersolanol (C16H20O7) and fluorinated derivatives are calculated using density functional theory (DFT) and molecular docking approaches. It is shown that the pure C16H20O7 molecule has a direct HOMO-LUMO energy gap about 3.1 eV. The substitution of F atom at A category decreases the electronic energy gap, while it is constant at B category. In A category, the behavior of the pure molecule changed from insulator to semiconductor with various substitution of F atom. The electronic properties were depended on the F sites in the pure molecule. The molecular electrical transport properties and charge-transfer possibilities increase with decreasing energy gap. The pure C16H20O7 molecule with high energy gap has low chemical reactivity and substitution of F atom at considered molecule increases chemical reactivity. Obtained results show that F-O bonds in trifurcation bonds of C16H19O7(F14), C16H19O7(F16), and C16H19O7(F17) molecules play a key role in confronting with COVID-19, HIV, and HTLV proteases, respectively. Optical spectra, such as the dielectric functions, electron energy-loss spectroscopy, refractive index, extinction coefficient, and reflection spectra show that fluorinated derivatives of C16H20O7 at B category can be used in the new drugs.


Summary of the clinical practice guideline for the treatment of posttraumatic stress disorder (PTSD) in adults.
The American Psychological Association (APA) developed a clinical practice guideline (CPG) to provide recommendations on psychological and pharmacological treatments for posttraumatic stress disorder (PTSD) in adults. This paper is a summary of the CPG, including the development process. Members of the guideline development panel (GDP) used a comprehensive systematic review conducted by the Research Triangle Institute-University of North Carolina Evidence-based Practice Center (RTI-UNC EPC) as its primary evidence base (Jonas et al., 2013). The GDP consisted of health professionals from psychology, psychiatry, social work, and family medicine as well as community members who self-identified as having had PTSD. PTSD symptom reduction and serious harms were selected by the GDP as critical outcomes for making recommendations. The GDP strongly recommends use of the following psychotherapies/interventions (in alphabetical order) for adults with PTSD: cognitive-behavioral therapy, cognitive processing therapy, cognitive therapy, and prolonged exposure therapy. The GDP conditionally recommends the use of brief eclectic psychotherapy, eye movement desensitization and reprocessing (EMDR), and narrative exposure therapy (NET). For medications, the GDP conditionally recommends the following (in alphabetical order): fluoxetine, paroxetine, sertraline, and venlafaxine. There is insufficient evidence to recommend for or against offering Seeking Safety, relaxation, risperidone, and topiramate. A subgroup of the GDP reviewed studies published after the systematic review for those treatments that received substantive recommendations; the GDP concluded that future systematic reviews that incorporated those new studies could change the recommendations for EMDR and NET from conditional to strong. For all other treatments, results of the update indicated that recommendations were unlikely to change or that there were no new trials for comparison. The target audience for this CPG includes clinicians, researchers, patients, and policymakers. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Management of red cell alloimmunisation in pregnancy: the non-invasive monitoring of the disease.
Haemolytic disease of the fetus and newborn (HDFN) due to red cell alloimmunization was a significant cause of fetal and neonatal morbidity and mortality until the introduction of anti-D immunoglobulin, which has dramatically changed the incidence of the disease. However, it is still a major problem in affected pregnancies. The emphasis of current clinical management has shifted from an invasive approach to non-invasive monitoring of the disease. The key elements of the modern management are determining which fetuses are at risk of HDFN with the use of cell-free fetal DNA in maternal plasma (fetal RHD genotype) and the follow-up of antigen positive fetuses by Doppler ultrasonography to detect anaemia severe enough to need treatment. When anaemia is suspected, an invasive approach is still required in a timely manner for confirmation of the degree of anaemia and to administer blood transfusions. This non-invasive approach prevents unnecessary administration of human-derived blood products, with the consequent ethical and cost implications and most importantly avoids iatrogenic conversion of mild to severe disease by avoiding need for techniques such as amniocentesis. The potential problem of the non-invasive approach is the reduction in the total number of invasive procedures, with the subsequent difficulty of maintaining the skills required to perform them.


Caffeic acid phenethyl ester rescued streptozotocin-induced memory loss through PI3-kinase dependent pathway.
The present study was undertaken to elucidate the role of PI3-kinase signaling in memory enhancing potential of caffeic acid phenethyl ester (CAPE) against cognitive defects in rats after centrally administered streptozotocin as a model of Alzheimer's disease. The Morris water maze and elevated plus maze paradigms showed profound loss of memory in adult Wistar rats (180-200 g) injected with streptozotocin (3 mg/kg) bilaterally (STZ-ICV) on day 1 and 3. Intraperitoneal administration of CAPE (6 mg/kg, i.p., 28 days) attenuated STZ-ICV triggered memory loss in rats. Treatment with PI3-kinase inhibitor (wortmannin, 5 μg/rat, ICV) or NOS blocker (L-NAME, 20 mg/kg, i.p., 28 days) interfered with memory restorative function of CAPE in STZ treated rats. In biochemical analysis markers of oxidative stress (TBARS, GSH, SOD, CAT), nitrite, AChE, TNF-α, eNOS and NFκB were measured in brain of rats on day 28. Interestingly, L-Arginine (100 mg/kg, i.p., 28 days) group exhibited moderate (p > 0.05) decline in memory functions. The brain oxidative stress, TNF-α, AChE activity and NFκB levels were elevated, and eNOS level was lowered by STZ-ICV treatment. Administration of CAPE lowered oxidative stress, AChE, nitrite and TNF-α levels in brain of rats. The eNOS level was enhanced and NFκB level was decreased by CAPE in STZ treated rats. Wortmannin injection elevated the brain oxidative stress, AChE activity and TNF-α levels, and decreased the nitrite, eNOS and NFκB level. Rise of brain oxidative stress parameters, AChE activity, TNF-α, eNOS and NFκB levels, and decline in brain nitrite content was observed in L-NAME treated group. L-Arginine administration showed modest effects (p > 0.05) on oxidative stress parameters. Brain nitrite content was enhanced although eNOS, NFκB levels, and AChE activity was decimated by L-Arginine treatment. It can be concluded that PI3-kinase mediated nitric oxide facilitation is an essential feature of CAPE action in STZ-ICV treated rats.


Broken ring solitons in Bessel optical lattices.
We address the existence of ring solitons broken by several nodes in a defocusing saturable nonlinear medium with an imprinted Bessel optical lattice. Such a multipolelike soliton is composed of two or more arc patterns with opposite phase between the adjacent components. The width of existence domain is determined only by the saturation degree of medium. The maximum number of soliton components depends on the radius of the lattice ring, where they reside. Those novel solitons can be trapped entirely on any ring of the Bessel lattice provided that the lattice is modulated deep enough. This study offers a smooth transition from the multipole soliton to necklace soliton.


Retinal vessel oxygen saturation increases after vitrectomy.
To evaluate the effects of vitrectomy on retinal vascular oxygen saturation in an adult population. This was a prospective observational study. Twenty-seven eyes of 27 patients who underwent vitrectomy for macular conditions were included. Retinal oximetry was performed using the Oxymap system prior to vitrectomy and 3 months after surgery and the mean retinal arterial and venous oxygen saturation were measured. The arterial-venous difference (AVD) was calculated as the difference between the arterial and venous saturations. Multivariate linear regression models were constructed to compare oxygen saturation before and after surgery, with adjustments for age, sex, hypertension, hyperlipidemia, diabetes mellitus, and indication for surgery. The mean age of the subjects was 68.4 ± 8.9 years, 15 (55%) were male and the majority were of Chinese ethnicity (93%). The mean arterial saturation increased significantly after vitrectomy (101.93 ± 8.36% vs. 96.16 ± 14.14%, P = 0.01). The mean venous saturation also increased significantly after surgery (59.76 ± 8.52% vs. 50.40 ± 11.72%, P = 0.02). The mean AVD significantly decreased from 45.76 ± 12.18% before surgery to 42.17 ± 10.94% after surgery (P = 0.02). Retinal arterial and venous oxygen saturation are significantly increased after vitrectomy, while the AVD is decreased after vitrectomy.


Intrapulse depolarization in optical fibers: a classical analog of spin decoherence.
We show that the combination of cross-phase modulation and polarization-mode dispersion inside optical fibers leads to a novel phenomenon of intrapulse depolarization manifested as different random states of polarization across the pulse profile. Such polarization evolution of optical pulses is directly analogous to the phenomenon of spin decoherence in semiconductors or pseudospin relaxation in atoms.


dmPFC-vlPAG projection neurons contribute to pain threshold maintenance and antianxiety behaviors.
The dorsal medial prefrontal cortex (dmPFC) has been recognized as a key cortical area for nociceptive modulation. However, the underlying neural pathway and the function of specific cell types remain largely unclear. Here, we show that lesions in the dmPFC induced an algesic and anxious state. Using multiple tracing methods including a rabies-based transsynaptic tracing method, we outlined an excitatory descending neural pathway from the dmPFC to the ventrolateral periaqueductal gray (vlPAG). Specific activation of the dmPFC/vlPAG neural pathway by optogenetic manipulation produced analgesic and antianxiety effects in a mouse model of chronic pain. Inhibitory neurons in the dmPFC were specifically activated using a chemogenetic approach, which logically produced an algesic and anxious state under both normal and chronic pain conditions. Antagonists of the GABAA receptor (GABAAR) or mGluR1 were applied to the dmPFC, which produced analgesic and antianxiety effects. In summary, the results of our study suggest that the dmPFC/vlPAG neural pathway might participate in the maintenance of pain thresholds and antianxiety behaviors under normal conditions, while silencing or suppressing the dmPFC/vlPAG pathway might be involved in the initial stages and maintenance of chronic pain and the emergence of anxiety-like behaviors.


Ancestral Folate Promotes Neuronal Regeneration in Serial Generations of Progeny.
Folate supplementation in F0 mating rodents increases regeneration of injured spinal axons in vivo in 4 or more generations of progeny (F1-F4) in the absence of interval folate administration to the progeny. Transmission of the enhanced regeneration phenotype to untreated progeny parallels axonal growth in neuron culture after in vivo folate administration to the F0 ancestors alone, in correlation with differential patterns of genomic DNA methylation and RNA transcription in treated lineages. Enhanced axonal regeneration phenotypes are observed with diverse folate preparations and routes of administration, in outbred and inbred rodent strains, and in two rodent genera comprising rats and mice, and are reversed in F4-F5 progeny by pretreatment with DNA demethylating agents prior to phenotyping. Uniform transmission of the enhanced regeneration phenotype to progeny together with differential patterns of DNA methylation and RNA expression is consistent with a non-Mendelian mechanism. The capacity of an essential nutritional co-factor to induce a beneficial transgenerational phenotype in untreated offspring carries broad implications for the diagnosis, prevention, and treatment of inborn and acquired disorders.


Intake of Caffeinated Soft Drinks before and during Pregnancy, but Not Total Caffeine Intake, Is Associated with Increased Cerebral Palsy Risk in the Norwegian Mother and Child Cohort Study.
Postnatal administration of caffeine may reduce the risk of cerebral palsy (CP) in vulnerable low-birth-weight neonates. The effect of antenatal caffeine exposure remains unknown. We investigated the association of intake of caffeine by pregnant women and risk of CP in their children. The study was based on The Norwegian Mother and Child Cohort Study, comprising >100,000 live-born children, of whom 222 were subsequently diagnosed with CP. Mothers reported their caffeine consumption in questionnaires completed around pregnancy week 17 (102,986 mother-child pairs), week 22 (87,987 mother-child pairs), and week 30 (94,372 mother-child pairs). At week 17, participants were asked about present and prepregnancy consumption. We used Cox regression models to estimate associations between exposure [daily servings (1 serving = 125 mL) of caffeinated coffee, tea, and soft drinks and total caffeine consumption] and CP in children, with nonconsumers as the reference group. Models included adjustment for maternal age and education, medically assisted reproduction, and smoking, and for each source of caffeine, adjustments were made for the other sources. Total daily caffeine intake before and during pregnancy was not associated with CP risk. High consumption (≥6 servings/d) of caffeinated soft drinks before pregnancy was associated with an increased CP risk (HR: 1.9; 95% CI: 1.2, 3.1), and children of women consuming 3-5 daily servings of caffeinated soft drinks during pregnancy weeks 13-30 also had an increased CP risk (HR: 1.7; 95% CI: 1.1, 2.8). A mean daily consumption of 51-100 mg caffeine from soft drinks during the first half of pregnancy was associated with a 1.9-fold increased risk of CP in children (HR: 1.9; 95% CI: 1.1, 3.6). Maternal total daily caffeine consumption before and during pregnancy was not associated with CP risk in children. The observed increased risk with caffeinated soft drinks warrants further investigation.


Successful Prediction of a Left Nonrecurrent Laryngeal Nerve in a Patient With Right-Sided Aorta and Aberrant Left Subclavian Artery.
Left nonrecurrent laryngeal nerve (LNRLN) is an extremely rare anatomic variant. The development of such anatomic variation requires the regression of both the fourth (aortic arch) and sixth (ductus arteriosus, DA) arches on the left side. Preoperative prediction of this variant is difficult but might reduce risk of nerve injury. A 34-year-old female was indicated for thyroidectomy for a 2.4 cm follicular neoplasm and Graves' disease. Due to a positive medical history of 22q11.2 microdeletion and unexplained left vocal cord paralysis, a preoperative chest computed tomography (CT) scan was obtained and revealed a right-sided aorta (RSA) and aberrant left subclavian artery (ALSA) without Kommerell's diverticulum. A left-sided NRLN was then highly suspected. Thyroidectomy was performed under general anesthesia with the utilization of intraoperative laryngeal nerve monitoring. A LNRLN was confirmed intraoperatively. Right-sided aorta and ALSA indicate embryologic regression of the left fourth primitive aortic arch. The absence of Kommerell's diverticulum at the origin of the ALSA indicates the lack of high-pressure blood flow from the pulmonary artery to the ALSA through the ductus arteriosus during embryogenesis, suggesting the embryologic regression of the left sixth primitive aortic arch. The presence of all 3 radiologic features thus highly suggests the possibility of a LNRLN.


Tailored medicine: whom will it fit? The ethics of patient and disease stratification.
A key selling point of pharmacogenetics is the genetic stratification of either patients or diseases in order to target the prescribing of medicine. The hope is that genetically 'tailored' medicines will replace the current 'one-size-fits-all' paradigm of drug development and usage. This paper is concerned with the relationship between difference and justice in the use of pharmacogenetics. This new technology, which facilitates the identification and use of difference, has, we shall argue, the potential to lead to injustice either by the inappropriate use of difference or through the inappropriate failure to use difference. We build on empirical data from a detailed study of the range of options for the development of pharmacogenetics to present a consideration of the ethical issues that surround patient and disease stratification. In it we explore the ways in which the use of pharmacogenetics may lead to the creation of new, genetically stratified, forms of difference and new forms of injustice based on these divisions. We also examine the ways in which existing forms of difference and social stratification may interact with the use of pharmacogenetics. In conclusion, we suggest how an understanding of these ethical issues could usefully inform future policy discussions.


Genetic Risk for Aortic Aneurysm in Adolescent Idiopathic Scoliosis.
Scoliosis is a feature of several genetic disorders that are also associated with aortic aneurysm, including Marfan syndrome, Loeys-Dietz syndrome, and type-IV Ehlers-Danlos syndrome. Life-threatening complications of aortic aneurysm can be decreased through early diagnosis. Genetic screening for mutations in populations at risk, such as patients with adolescent idiopathic scoliosis, may improve recognition of these disorders. The coding regions of five clinically actionable genes associated with scoliosis (COL3A1, FBN1, TGFBR1, TGFBR2, and SMAD3) and aortic aneurysm were sequenced in 343 adolescent idiopathic scoliosis cases. Gene variants that had minor allele frequencies of <0.0001 or were present in human disease mutation databases were identified. Variants were classified as pathogenic, likely pathogenic, or variants of unknown significance. Pathogenic or likely pathogenic mutations were identified in 0.9% (three) of 343 adolescent idiopathic scoliosis cases. Two patients had pathogenic SMAD3 nonsense mutations consistent with type-III Loeys-Dietz syndrome and one patient had a pathogenic FBN1 mutation with subsequent confirmation of Marfan syndrome. Variants of unknown significance in COL3A1 and FBN1 were identified in 5.0% (seventeen) of 343 adolescent idiopathic scoliosis cases. Six FBN1 variants were previously reported in patients with Marfan syndrome, yet were considered variants of unknown significance based on the level of evidence. Variants of unknown significance occurred most frequently in FBN1 and were associated with greater curve severity, systemic features of Marfan syndrome, and joint hypermobility. Clinically actionable pathogenic mutations in genes associated with adolescent idiopathic scoliosis and aortic aneurysm are rare in patients with adolescent idiopathic scoliosis who are not suspected of having these disorders, although variants of unknown significance are relatively common. Routine genetic screening of all patients with adolescent idiopathic scoliosis for mutations in clinically actionable aortic aneurysm disease genes is not recommended on the basis of the high frequency of variants of unknown significance. Clinical evaluation and family history should heighten indications for genetic referral and testing.


Recombination and the frequency spectrum in Drosophila melanogaster and Drosophila simulans.
Most "tests of neutrality" assess whether particular data sets depart from the predictions of a standard neutral model with no recombination. For Drosophila, where nuclear polymorphism data routinely show evidence of genetic exchange, the assumption of no recombination is often unrealistic. In addition, while conservative, this assumption is made at the cost of a great loss in power. Perhaps as a result, tests of the frequency spectrum based on zero recombination suggest an adequate fit of Drosophila polymorphism data to the predictions of the standard neutral model. Here, we analyze the frequency spectrum of a large number of loci in Drosophila melanogaster and D. simulans using two summary statistics. We use an estimate of the population recombination rate based on a laboratory estimate of the rate of crossing over per physical length and an estimate of the species' effective population size. In contrast to previous studies, we find that roughly half of the loci depart from the predictions of the standard neutral model. The extent of the departure depends on the exact recombination rate, but the global pattern that emerges is robust. Interestingly, these departures from neutral expectations are not unidirectional. The large variance in outcomes may be due to a complex demographic history and inconsistent sampling, or to the pervasive action of natural selection.


Abnormal whole-brain functional networks in homogeneous acute mild traumatic brain injury.
To evaluate the whole-brain resting-state networks in a homogeneous group of patients with acute mild traumatic brain injury (MTBI) and to identify alterations in functional connectivity induced by MTBI. Thirty-five patients with acute MTBI and 35 healthy control subjects, matched in age, gender, handedness, and education, underwent resting-state fMRI, susceptibility weighted imaging, neuropsychological, and postconcussive symptom assessments. We ensured the homogeneity of the patient group by limiting the injury mechanism to fronto-occipital impacts. Alterations in functional connectivity were analyzed by using data-driven independent component analysis, which is not biased by a priori region selection. We found a decrease in functional connectivity within the motor-striatal network in the MTBI group. At the same time, patients showed deficits in psychomotor speed as well as in speed of information processing. We propose that although disorders in motor function after MTBI are rarely reported, injury still has an effect on motor functioning, which in its turn may also explain the reduction in speed of information processing. Further, we found a cluster of increased functional connectivity in the right frontoparietal network in the MTBI group. We suggest that this abnormal increased connectivity might reflect increased awareness to external environment and explain excessive cognitive fatigue reported by patients with MTBI. It might also underlie the physical postconcussive symptoms, such as headache and increased sensitivity to noise/light. We proved that whole-brain functional connectivity is altered early (within 4 weeks) after MTBI, suggesting that changes in functional networks underlie the cognitive deficits and postconcussive complaints reported by patients with MTBI.


Developmental and behavioral disorders grown up: attention deficit hyperactivity disorder.
Pat was diagnosed with attention deficit hyperactivity disorder-combined type in the second grade and has been treated primarily with stimulants, albeit on an inconsistent basis. Pat is now a 22-year-old male, who is a second year student at a local community college. He attained a 2.1 GPA during his first year of postsecondary studies, doing quite well in several classes yet rather poorly in several others. Pat has a part-time job at a fast food restaurant, a position that he has held for approximately 6 months. Before his current job, Pat worked as a dishwasher at a local restaurant. He was fired from this position due to chronically being late to work. Pat stated that he gets along well with his peers, although his mother reported that Pat has a history of being able to make friends, "yet not keep them." Pat's mother also reported that Pat has traditionally gravitated toward peers who were "not especially good influences." She described various antisocial acts that Pat was involved with during high school. Pat reported that he began experimenting with alcohol and cigarettes in ninth grade and marijuana shortly thereafter. He stated that he drinks less often than he smokes marijuana ("several times a week"). Pat denied legal difficulties, although his mother reported that Pat was asked to leave the dormitory during his first semester due to an alcohol-related incident. Pat is not currently involved in psychotherapy and is prescribed extended release methylphenidate. He reported that he took his stimulant whenever he had to do a lot of work for school.


Characterization of gelatinases in bronchoalveolar lavage fluid and gelatinases produced by alveolar macrophages isolated from healthy calves.
To characterize gelatinases in bronchoalveolar lavage fluid (BALF) and gelatinases produced by alveolar macrophages of healthy calves. Samples of BALF and alveolar macrophages obtained from 20 healthy 2-month-old calves. BALF was examined by use of gelatin zymography and immunoblotting to detect gelatinases and tissue inhibitor of metalloproteinase (TIMP)-1 and -2. Cultured alveolar macrophages were stimulated with lipopolysaccharide (LPS), and conditioned medium was subjected to zymography. Alveolar macrophage RNA was used for reverse transcriptase-polymerase chain reaction assay of matrix metalloproteinases (MMPs), cyclooxygenase-2, and inducible nitric oxide synthase. Gelatinolytic activity in BALF was evident at 92 kd (14/20 calves; latent MMP-9) and 72 kd (18/20; latent MMP-2). Gelatinolytic activity was evident at 82 kd (10/20 calves; active MMP-9) and 62 kd (17/20; active MMP-2). Gelatinases were inhibited by metal chelators but not serine protease inhibitors. Immunoblotting of BALF protein and conditioned medium confirmed the MMP-2 and -9 proteins. Endogenous inhibitors (ie, TIMPs) were detected in BALF from all calves (TIMP-1) or BALF from only 4 calves (TIMP-2). Cultured alveolar macrophages expressed detectable amounts of MMP-9 mRNA but not MMP-2 mRNA. Healthy calves have detectable amounts of the gelatinases MMP-2 and -9 in BALF Endogenous inhibitors of MMPs were detected in BALF (ie, TIMP-1, all calves; TIMP-2, 4 calves). Lipopolysaccharide-stimulated alveolar macrophages express MMP-9 but not MMP-2 mRNA. The role of proteases in the pathogenesis of lung injury associated with pneumonia has yet to be determined.


INFANT AVOIDANCE DURING A TACTILE TASK PREDICTS AUTISM SPECTRUM BEHAVIORS IN TODDLERHOOD.
The experience of touch is critical for early communication and social interaction; infants who show aversion to touch may be at risk for atypical development and behavior problems. The current study aimed to clarify predictive associations between infant responses to tactile stimuli and toddler autism spectrum, internalizing, and externalizing behaviors. This study measured 9-month-old infants' (N = 561; 58% male) avoidance and negative affect during a novel tactile task in which parents painted infants' hands and feet and pressed them to paper to make a picture. Parent reports on the Pervasive Developmental Problems (PDP), Internalizing, and Externalizing scales of the Child Behavior Checklist were used to measure toddler behaviors at 18 months. Infant observed avoidance and negative affect were significantly correlated; however, avoidance predicted subsequent PDP scores only, independent of negative affect, which did not predict any toddler behaviors. Findings suggest that incorporating measures of responses to touch in the study of early social interaction may provide an important and discriminating construct for identifying children at greater risk for social impairments related to autism spectrum behaviors.