Metabolites associated with adaptation of microorganisms to an acidophilic, metal-rich environment identified by stable-isotope-enabled metabolomics.
Microorganisms grow under a remarkable range of extreme conditions. Environmental transcriptomic and proteomic studies have highlighted metabolic pathways active in extremophilic communities. However, metabolites directly linked to their physiology are less well defined because metabolomics methods lag behind other omics technologies due to a wide range of experimental complexities often associated with the environmental matrix. We identified key metabolites associated with acidophilic and metal-tolerant microorganisms using stable isotope labeling coupled with untargeted, high-resolution mass spectrometry. We observed >3,500 metabolic features in biofilms growing in pH ~0.9 acid mine drainage solutions containing millimolar concentrations of iron, sulfate, zinc, copper, and arsenic. Stable isotope labeling improved chemical formula prediction by >50% for larger metabolites (>250 atomic mass units), many of which were unrepresented in metabolic databases and may represent novel compounds. Taurine and hydroxyectoine were identified and likely provide protection from osmotic stress in the biofilms. Community genomic, transcriptomic, and proteomic data implicate fungi in taurine metabolism. Leptospirillum group II bacteria decrease production of ectoine and hydroxyectoine as biofilms mature, suggesting that biofilm structure provides some resistance to high metal and proton concentrations. The combination of taurine, ectoine, and hydroxyectoine may also constitute a sulfur, nitrogen, and carbon currency in the communities. Microbial communities are central to many critical global processes and yet remain enigmatic largely due to their complex and distributed metabolic interactions. Metabolomics has the possibility of providing mechanistic insights into the function and ecology of microbial communities. However, our limited knowledge of microbial metabolites, the difficulty of identifying metabolites from complex samples, and the inability to link metabolites directly to community members have proven to be major limitations in developing advances in systems interactions. Here, we show that combining stable-isotope-enabled metabolomics with genomics, transcriptomics, and proteomics can illuminate the ecology of microorganisms at the community scale.


An HPLC-MS method for simultaneous estimation of alpha,beta-arteether and its metabolite dihydroartemisinin, in rat plasma for application to pharmacokinetic study.
This manuscript reports, the development and validation of a sensitive and selective assay method for simultaneous determination of alpha,beta-arteether and its metabolite dihydroartemisinin (DHA) in rat plasma by liquid chromatography-mass spectrometry. Chromatographic separations were achieved by gradient elution of the analytes with an initial composition of methanol-potassium acetate buffer (pH 4; 73:27, v/v) to 100% methanol in 3 min and maintained for 5 min on a Spheri-10, RP(18) (100 x 4.6 mm i.d.) column following an RP(18) (30 x 4.6 mm i.d.) guard column. The total effluent from the column was split so that one-tenth was injected into the electrospray LC/MS interface. ESI-MS analysis was performed using a Micromass Quattro II Triple Quadrupole Mass Spectrometer equipped with an electrospray source. The MS analysis was carried out at cone voltage of 22 V with a scan range of 200-500 Da. The analytes were quantified from the [M+ K](+) ion chromatograms of alpha,beta-arteether at m/z 352, DHA at m/z 323, artemisinin at m/z 321 and propyl ether analogue of arteether at m/z 365. Liquid-liquid extractions with a combination of n-hexane and hexane-ethyl acetate (8:2) were used to isolate alpha,beta-arteether and DHA from rat plasma. The method was validated and gave good accuracy and precision for the studied domain. Linearity in serum was observed over the range 4.375-70 ng/mL for alpha-arteether and 10-160 ng/mL for beta-arteether and DHA. Percentage bias (accuracy) and within- and between-assay precision were well within the acceptable range. This method was applied to study the pharmacokinetics following oral administration of alpha,beta-arteether (30 mg/kg) in rats.


The role of 131 iodine-metaiodobenzylguanidine scanning in the correlative imaging of patients with neuroblastoma.
Metaiodobenzylguanidine (MIBG) scans were studied to determine the impact of the scan results on the clinical treatment of pediatric patients with neural crest tumors. Serial scans were reviewed retrospectively for 27 patients with neural crest tumors: 25 with initial diagnoses of neuroblastoma (NB), 1 with ganglioneuroblastoma, and 1 with ganglioneuroma (GN). Results were compared with bone scans and computed tomography scans, as well as surgical pathologic findings. At initial diagnosis, when compared with bone and computed tomographic scans, MIBG imaging did not identify any unsuspected lesions that resulted in a change in staging. Thirteen patients with NB who had initially positive MIBG scan results had serial studies that normalized during therapy. However, after completion of therapy, 8 of 13 had relapses of the disease. Although areas of active disease were well delineated by other standard imaging modalities for all 8, only 4 (50%) had MIBG study results that were positive in sites of relapse. There were 4 cases of GN (1 at diagnosis and 3 after therapy for NB) demonstrating an uptake of MIBG that was similar in appearance to that in NB. MIBG imaging did not change the staging or alter treatment during therapy for any patient. Normalization of positive study results was an unreliable indicator of outcome for children with NB. Furthermore, when relapse occurred, MIBG scans identified only 50% of those with active NB. The uptake of MIBG in GN was indistinguishable from that in NB. In this series, the results of serial MIBG studies did not have a significant impact on patient treatment.


Evaluation of two phenanthrenemethanols for antimalarial activity in man: WR 122,455 and WR 171,669.
Two phenanthrenemethanols, WR 122,455 and WR 171,669, were tested in man for oral tolerance, toxicity, and efficacy against Plasmodium falciparum. In healthy subjects, gastrointestinal symptoms limited single-day dosage of WR 122,455 to 800 mg and WR 171,669 to 1,260 mg. No laboratory abnormalities phototoxicity, or gastrointestinal blood loss were noted in the subjects receiving these drugs. In subjects infected with malaria, WR 122,455 cleared parasitemia when given for 1 day in doses from 440 to 880 mg; recrudescences occurred in these subjects who were infected with either chloroquine-sensitive or chloroquine-resistant P. falciparum. However, WR 122,455 at 480 mg/day for 3 to 6 days cured 9/9 cases of chloroquine-resistant and 4/4 cases of chloroquine-sensitive P. falciparum infection. WR 171,669 at 1 g/day for 3 days cured 6/6 subjects with chloroquine-resistant and 3/3 cases of chloroquine-sensitive infection. Both agents cleared parasitemia and fever promptly.


Purification and properties of myo-inositol-1-phosphatase from bovine brain.
myo-Inositol-1-phosphatase from bovine brain was purified over 2000-fold. The native enzyme has a Mr of 59,000, and on SDS/polyacrylamide-gel electrophoresis the subunit Mr was 31,000. Thus the native enzyme is a dimer of two apparently identical subunits. The enzyme, purified to a specific activity of more than 300 units/mg of protein (1 unit of enzyme activity corresponds to the release of 1 mumol of Pi/h at 37 degrees C), catalysed the hydrolysis of a variety of phosphorylated compounds, the best one, in terms of V/Km, being D-myo-inositol 1-phosphate. Kinetic constants of compounds tested, including both isomers of glycerophosphate and two deoxy forms of beta-glycerophosphate, were measured. They show the importance of the two hydroxyl groups which are adjacent to the phosphate in myo-inositol 1-phosphate. With a wide variety of substrates Li+ was found to be an uncompetitive inhibitor whose Ki varied with substrate structure.


Electrophysiological, biochemical, and bioinformatic methods for studying CFTR channel gating and its regulation.
CFTR is the only member of the ABC (ATP-binding cassette) protein superfamily known to function as an ion channel. Most other ABC proteins are ATP-driven transporters, in which a cycle of ATP binding and hydrolysis, at intracellular nucleotide binding domains (NBDs), powers uphill substrate translocation across the membrane. In CFTR, this same ATP-driven cycle opens and closes a transmembrane pore through which chloride ions flow rapidly down their electrochemical gradient. Detailed analysis of the pattern of gating of CFTR channels thus offers the opportunity to learn about mechanisms of function not only of CFTR channels but also of their ABC transporter ancestors. In addition, CFTR channel gating is subject to complex regulation by kinase-mediated phosphorylation at multiple consensus sites in a cytoplasmic regulatory domain that is unique to CFTR. Here we offer a practical guide to extract useful information about the mechanisms that control opening and closing of CFTR channels: on how to plan (including information obtained from analysis of multiple sequence alignments), carry out, and analyze electrophysiological and biochemical experiments, as well as on how to circumvent potential pitfalls.


Association between continued residence in temporary prefabricated housing and musculoskeletal pain in survivors of the Great East Japan Earthquake: a longitudinal study.
Prolonged periods of living in prefabricated houses (PHs) may increase the risk of musculoskeletal (MSK) symptoms; however, the association is not clear. This study aimed to investigate the association between continued residence in PHs and MSK pain in a population affected by a natural disaster, the Great East Japan Earthquake (GEJE) survivors. A panel study was conducted including 1059 and 792 survivors at 2 and 4 years, respectively, after the GEJE, using a self-reported questionnaire. Those with no response on living status and those who did not live in a PH were excluded. Participants were classified into two groups by living status: continued residence in a PH (lived in a PH during both periods) or moving out of a PH (lived in a PH in the first period and did not live in a PH in the second). MSK pain included lower back, shoulder, knee, hand or foot, and neck pain. Changes in the occurrence of MSK pain during the two periods were assessed and defined as 'new-onset' and 'continuing' MSK pain. Multiple logistic regression analysis was used to examine the influence of continued residence in a PH on new-onset and continuing MSK pain. Continued residence in a PH was significantly associated with new-onset MSK pain, even after adjustment for covariates (adjusted OR 2.18, 95% CI 1.25 to 3.79, p=0.006). Participants who continued living in a PH had higher rates of continuing MSK pain than those who moved out; however, the difference was not significant (adjusted OR 1.69, 95% CI 0.94 to 3.05, p=0.079). Continued residence in a PH was associated with new-onset MSK pain among survivors. Public support should be provided to such people to ensure a more comfortable life.


Fabrication of hydrogel-coated single conical nanochannels exhibiting controllable ion rectification characteristics.
Heterogeneous nanochannel materials that endow new functionalities different to the intrinsic properties of two original nanoporous materials have wide potential applications in nanofluidics, energy conversion, and biosensors. Herein, we report novel, interesting hydrogel-composited nanochannel devices with regulatable ion rectification characteristics. The heterogeneous nanochannel devices were constructed by selectively coating the tip side, base side, or both sides of a single conical nanochannel membrane with thin agar hydrogel layers. The tunable ion current rectification of the nanochannels in the three different coating states was systematically demonstrated by current-voltage (I-V) curves. The asymmetric ionic transport property of the conical nanochannel was further strengthened in the tip-coating state and weakened in the base-coating state, whereas the conical nanochannel showed nearly symmetric ionic transport in the dual-coating state. Repeated experiments presented insight into the good stability and reversibility of the three coating states of the hydrogel-nanochannel-integrated systems. This work, as an example, may provide a new strategy to further design and develop multifunctional gel-nanochannel heterogeneous smart porous nanomaterials.


Circulating Tumor DNA: Towards More Individualized Treatment for Patients with Resectable Colorectal Cancer.
Despite curative-intent treatment, recurrence is common for patients with colorectal cancer (CRC). Currently, prediction of disease recurrence and prognostication following surgery is based upon vague clinical factors and more precise and dynamic biomarkers for risk stratification and treatment decisions are urgently needed. Circulating tumor DNA (ctDNA) is a promising biomarker for patients undergoing treatment for resectable CRC. In this review, we provide an overview of the data supporting current uses of ctDNA for CRC, including localized CRC and resectable colorectal liver metastases (CLM), as well as descriptions of important ongoing clinical trials using ctDNA in the care of patients with CRC. The detection of ctDNA following curative-intent therapy is associated with disease recurrence, and multiple trials are investigating its role in determining need and duration for adjuvant therapy for localized CRC. In addition, ctDNA reliably predicts prognosis for patients with CLM, with trials underway studying ctDNA-guided treatment sequencing and intensity. The detection of ctDNA is a sensitive and dynamic biomarker for disease recurrence in CRC. Many investigations are underway into ctDNA's potential role in surveillance and treatment algorithms, and it has the potential to become a critical biomarker to determine individualized strategies for treatment sequencing, choice, and duration of therapies.


Impact of anesthesia, analgesia, and euthanasia technique on the inflammatory cytokine profile in a rodent model of severe burn injury.
Anesthetics used in burn and trauma animal models may be influencing results by modulating inflammatory and acute-phase responses. Accordingly, we determined the effects of various anesthetics, analgesia, and euthanasia techniques in a rodent burn model. Isoflurane (ISO), ketamine-xylazine (KX), or pentobarbital (PEN) with or without buprenorphine were administered before scald-burn in 72 rats that were euthanized without anesthesia by decapitation after 24 h and compared with unburned shams. In a second experiment, 120 rats underwent the same scald-burn injury using KX, and 24 h later were euthanized under anesthesia or carbon dioxide (CO2). In addition, we compared euthanasia by exsanguination with that of decapitation. Serum cytokine levels were determined by an enzyme-linked immunosorbent assay. In the first experiment, ISO was associated with elevation of cytokine-induced neutrophil chemoattractant 2 (CINC-2) and monocyte chemotactic protein 1 (MCP-1), and KX and PEN was associated with elevation of CINC-1,CINC-2, IL-6, and MCP-1. Pentobarbital also decreased IL-1". IL-6 increased significantly when ISO or PEN were combined with buprenorphine. In the second experiment, euthanasia performed by exsanguination under ISO was associated with reduced levels of IL-1", CINC-1, CINC-2, and MCP-1, whereas KX reduced CINC-2 and increased IL-6 levels. Meanwhile, PEN reduced levels of IL-1" and MCP-1, and CO2 reduced CINC-2 and MCP-1. In addition,decapitation after KX, PEN, or CO2 decreased IL-1" and MCP-1, although we found no significant difference between ISO and controls. Euthanasia by exsanguination compared with decapitation using the same agent also led to modulation of several cytokines. Differential expression of inflammatory markers with the use of anesthetics and analgesics should be considered when designing animal studies and interpreting results because these seem to have a significant modulating impact. Our findings indicate that brief anesthesia with ISO immediately before euthanasia by decapitation exerted the least dampening effect on the cytokines measured. Conversely, KX with buprenorphine may offer a better balance during longer procedures to avoid significant modulation. Standardization across all experiments that are compared and awareness of these findings are essential for those investigating the pathophysiology of inflammation in animal models.


Affinity maturation of a broadly neutralizing human monoclonal antibody that prevents acute hepatitis C virus infection in mice.
Direct-acting antivirals (DAAs) have led to a high cure rate in treated patients with chronic hepatitis C virus (HCV) infection, but this still leaves a large number of treatment failures secondary to the emergence of resistance-associated variants (RAVs). To increase the barrier to resistance, a complementary strategy is to use neutralizing human monoclonal antibodies (HMAbs) to prevent acute infection. However, earlier efforts with the selected antibodies led to RAVs in animal and clinical studies. Therefore, we identified an HMAb that is less likely to elicit RAVs for affinity maturation to increase potency and, more important, breadth of protection. Selected matured antibodies show improved affinity and neutralization against a panel of diverse HCV isolates. Structural and modeling studies reveal that the affinity-matured HMAb mediates virus neutralization, in part, by inducing conformational change to the targeted epitope, and that the maturated light chain is responsible for the improved affinity and breadth of protection. A matured HMAb protected humanized mice when challenged with an infectious HCV human serum inoculum for a prolonged period. However, a single mouse experienced breakthrough infection after 63 days when the serum HMAb concentration dropped by several logs; sequence analysis revealed no viral escape mutation. The findings suggest that a single broadly neutralizing antibody can prevent acute HCV infection without inducing RAVs and may complement DAAs to reduce the emergence of RAVs. (Hepatology 2016;64:1922-1933).


[The influence of metoprolol eye drops on intraocular pressure (author's transl)].
The selective beta-1 blocker Metoprolol was tested for a period of one month in a double-blind trial involving 36 patients with open-angle glaucoma or intraocular hypertension in whom the substance was applied twice daily to the eye in concentrations of 1, 2, 4 and 8%. At all these concentrations the intraocular pressure showed a reduction of 23%. The concentrations over 1% were less well tolerated, the difference being statistically significant; the patients concerned complained of an unpleasant burning sensation for 30 seconds after the application. The subjective intolerance shown increased in proportion to the concentration. It is not clear whether the solvent used contributed to the intolerance. The Metoprolol drops had no effect on pupillary diameter, blood pressure or resting pulse. An important finding was that during the one month's treatment there was no fall-off in the effectiveness of the preparation, i.e., the reduction in intraocular pressure and duration of action showed no diminution such as is seen, for example, with Timolol.


Effect of hyperoxia and vitamin C on coronary blood flow in patients with ischemic heart disease.
Pathological formation of reactive oxygen species within the coronary circulation has been hypothesized to mediate some clinical manifestations of ischemic heart disease (IHD) by interfering with physiological regulation of coronary tone. To determine the degree to which coronary tone responds to acute changes in ambient levels of oxidants and antioxidants in vivo in a clinical setting, we measured the effect of an acute oxidative stress (breathing 100% oxygen) on coronary capacitance artery diameter (quantitative angiography) and blood flow velocity through the coronary microcirculation (intracoronary Doppler ultrasonography) before and after treatment with the antioxidant vitamin C (3-g intravenous infusion) in 12 IHD patients undergoing a clinical coronary interventional procedure. Relative to room air breathing, 100% oxygen breathing promptly reduced coronary blood flow velocity by 20% and increased coronary resistance by 23%, without significantly changing the diameter of capacitance arteries. Vitamin C administration promptly restored coronary flow velocity and resistance to a slightly suprabasal level, and it prevented the reinduction of coronary constriction with rechallenge with 100% oxygen. This suggests that acute oxidative stress produces prompt and substantial changes in coronary resistance and blood flow in a clinical setting in patients with IHD, and it suggests that these changes are mediated by vitamin C-quenchable substances acting on the coronary microcirculation. This observation may have relevance for clinical practice.


Calcium channels and their role in regenerative medicine.
Stem cells hold indefinite self-renewable capability that can be differentiated into all desired cell types. Based on their plasticity potential, they are divided into totipotent (morula stage cells), pluripotent (embryonic stem cells), multipotent (hematopoietic stem cells, multipotent adult progenitor stem cells, and mesenchymal stem cells [MSCs]), and unipotent (progenitor cells that differentiate into a single lineage) cells. Though bone marrow is the primary source of multipotent stem cells in adults, other tissues such as adipose tissues, placenta, amniotic fluid, umbilical cord blood, periodontal ligament, and dental pulp also harbor stem cells that can be used for regenerative therapy. In addition, induced pluripotent stem cells also exhibit fundamental properties of self-renewal and differentiation into specialized cells, and thus could be another source for regenerative medicine. Several diseases including neurodegenerative diseases, cardiovascular diseases, autoimmune diseases, virus infection (also coronavirus disease 2019) have limited success with conventional medicine, and stem cell transplantation is assumed to be the best therapy to treat these disorders. Importantly, MSCs, are by far the best for regenerative medicine due to their limited immune modulation and adequate tissue repair. Moreover, MSCs have the potential to migrate towards the damaged area, which is regulated by various factors and signaling processes. Recent studies have shown that extracellular calcium (Ca2+) promotes the proliferation of MSCs, and thus can assist in transplantation therapy. Ca2+ signaling is a highly adaptable intracellular signal that contains several components such as cell-surface receptors, Ca2+ channels/pumps/exchangers, Ca2+ buffers, and Ca2+ sensors, which together are essential for the appropriate functioning of stem cells and thus modulate their proliferative and regenerative capacity, which will be discussed in this review.


Specific Signatures of the Gut Microbiota and Increased Levels of Butyrate in Children Treated with Fermented Cow's Milk Containing Heat-Killed Lactobacillus paracasei CBA L74.
We recently demonstrated that cow's milk fermented with the probiotic Lactobacillus paracasei CBA L74 (FM-CBAL74) reduces the incidence of respiratory and gastrointestinal tract infections in young children attending school. This effect apparently derives from a complex regulation of non-immune and immune protective mechanisms. We investigated whether FM-CBAL74 could regulate gut microbiota composition and butyrate production. We randomly selected 20 healthy children (12 to 48 months) from the previous randomized controlled trial, before (t0) and after 3 months (t3) of dietary treatment with FM-CBAL74 (FM) or placebo (PL). Fecal microbiota was profiled using 16S rRNA gene amplicon sequencing, and the fecal butyrate concentration was also measured. Microbial alpha and beta diversities were not significantly different between groups prior to treatment. FM-CBAL74 but not PL treatment increased the relative abundance of Lactobacillus Individual Blautia, Roseburia, and Faecalibacterium oligotypes were associated with FM-CBAL74 treatment and demonstrated correlative associations with immune biomarkers. Accordingly, PICRUSt analysis predicted an increase in the proportion of genes involved in butyrate production pathways, consistent with an increase in fecal butyrate observed only in the FM group. Dietary supplementation with FM-CBAL74 induces specific signatures in gut microbiota composition and stimulates butyrate production. These effects are associated with changes in innate and acquired immunity.IMPORTANCE The use of a fermented milk product containing the heat-killed probiotic strain Lactobacillus paracasei CBAL74 induces changes in the gut microbiota, promoting the development of butyrate producers. These changes in the gut microbiota composition correlate with increased levels of innate and acquired immunity biomarkers.


Association of white matter hyperintensities with low serum 25-hydroxyvitamin D levels.
Vitamin D deficiency is associated with cognitive impairment in the elderly and with increased white matter T2 hyperintensities in elderly debilitated patients. We investigated the relationship between serum vitamin D and brain MR findings in adult outpatients. Brain MR studies of 56 patients ages 30-69 years were selected when vitamin D level had been obtained within 90 days of the MRI. White matter T2 hyperintensities were characterized by size and location by two neuroradiologists. Manual volumetric analysis was assessed in patients more than 50 years of age. The entire cohort showed a significant negative relationship between serum 25-hydroxyvitamin D and the number of confluent juxtacortical white matter T2 hyperintensities (P = .047). The cohort ages 50 years and older showed stronger correlation between confluent white matter T2 hyperintensities and serum 25-hydroxyvitamin D in the juxtacortical region; number (P = .015) and size of white matter T2 hyperintensities (P = .048). Atrophy was not significantly related to serum 25-hydroxyvitamin D by radiologist visual analysis or by the bicaudate ratio. We found a significant relationship between vitamin D and white matter T2 hyperintensities in independent adult outpatients, especially over the age of 50 years.


Cognitive behavioural therapy and mindfulness based stress reduction may be equally effective in reducing anxiety and depression in adults with autism spectrum disorders.
Anxiety and depression co-occur in 50-70% of adults with autism spectrum disorder (ASD) but treatment methods for these comorbid problems have not been systematically studied. Recently, two ASD-tailored protocols were published: mindfulness based stress reduction (MBSR) and cognitive behavioural therapy (CBT). We wanted to investigate if both methods are equally effective in reducing anxiety and depression symptoms among adults with ASD. 59 adults with ASD and anxiety or depression scores above 7 on the Hospital Anxiety and Depression Scale, gave informed consent to participate; 27 followed the CBT protocol, and 32 the MBSR treatment protocol. Anxiety and depression scores, autism symptoms, rumination, and global mood were registered at the start, at the end of the 13-week treatment period, and at 3-months follow-up. Irrational beliefs and mindful attention awareness were used as process measures during treatment and at follow-up. Results indicate that both MBSR and CBT are associated with a reduction in anxiety and depressive symptoms among adults with ASD, with a sustained effect at follow-up, but without a main effect for treatment group. A similar pattern was seen for the reduction of autistic symptoms, rumination and the improvement in global mood. There are some indications that MBSR may be preferred over CBT with respect to the treatment effect on anxiety when the scores on measures of irrational beliefs or positive global mood at baseline are high. Mindfulness and cognitive behavioral therapies are both promising treatment methods for reducing comorbid anxiety and depression in adults with ASD.


Host immune deficiency in immune complex nephritis.
Evidence is reviewed that indicates that an immune deficiency state may underlie patients with immune complex nephritis. Studies in several laboratories provide data that show defective immune function in patients with nephritis at the following loci: defective immunoglobulin production, impaired monocyte Fc and C3b receptor function, diminished complement biosynthesis, and decreased mesangial cell colony-stimulating factor-1 biosynthesis. Aberrant monocyte function may be an important common denominator. The validation of this hypothesis has important implications for therapeutic strategies.


An evaluation of test and removal for the elimination of porcine reproductive and respiratory syndrome virus from 5 swine farms.
The objective of this field study was to evaluate the protocol of test and removal (T&R) for the elimination of porcine reproductive and respiratory syndrome virus (PRRSV) from 5 chronically infected breeding herds. The T&R protocol involved sampling the entire breeding herd in one day, testing sera by polymerase chain reaction and ELISA to detect previously exposed and/or infected animals, and subsequently removing them from the herd. Following completion of T&R, breeding herds were monitored for 12 consecutive months, using ELISA, for the presence of antibodies to PRRSV. In order to be classified as a PRRSV-negative herd, all samples collected over the 12-month monitoring period were required to be negative by ELISA (s/p ratio < 0.4). At the conclusion of the monitoring period, all 5 farms were PRRSV-negative, according to the defined testing criteria. Approximately 2.2% (74/3408) ELISA false positive samples were detected across all 5 farms during the monitoring period. The diagnostic cost required during the T&R protocol was approximately US $10.66 per animal tested. Limitations of the study were a lack of herds with large (> 2000 sows) breeding herd inventories, and herds with a history of PRRSV vaccination.


Intermediate Skin Substitutes Are Unnecessary in Small (<10% TBSA) Burns.
The use of intermediate skin substitutes between debridement and final autografting is routine for many practitioners. Materials such as xenografts and allografts have been promoted to help with wound coverage before autografting. However, there is limited data for their use in relatively small burn wounds (<10% TBSA). In this study, we analyzed the outcomes of 100 consecutive patients who underwent autografting for burns <10% TBSA at our American Burn Association-verified burn unit in the absence of intermediate skin substitute use. We retrospectively analyzed 100 patients who underwent split thickness skin graft autografting for burns <10% TBSA between November 2017 and June 2019. No patients were treated with intermediate skin substitutes. Analysis included basic demographics, comorbidities, TBSA burned, mechanism of burn, time to grafting, if grafting was performed in a single procedure or staged, graft loss (>50% graft failure), and time to complete healing (no further wound care required). Twelve patients (12%) had unpredictable graft beds, and their procedure was staged. These patients underwent surgical debridement and were dressed in antimicrobial dressing for an average of 5 days before autografting. No patients had intermediate skin substitutes between procedures. Eighty-eight patients (88%) were debrided and grafted in a single stage. In the staged group, there was a 0% rate of graft failure compared with 9.1% rate of graft failure in the primarily grafted group (P = .004). There was a similar length of stay and time to complete healing in the staged group and primarily grafted group (P = .496 and P = .571). There was a significantly shorter time from injury to first procedure between the staged group and the primarily grafted group (8.7 days and 13.5 days, P = .014). In the eight instances of graft failure, infection or inadequate debridement was the cause. Seven of these eight cases required further surgical intervention. Intermediate skin substitutes are an unnecessary step in grafting small burns. These add only complexity and cost to patient care. Many patients can be debrided and grafted in a single stage. Debridement alone with delayed grafting is a highly effective surgical method when the wound bed is not suitable for immediate grafting. The use of intermediate skin substitutes in small burns requires further investigation as this study finds low benefit for this product.


Evaluation of a Centyrin-Based Near-Infrared Probe for Fluorescence-Guided Surgery of Epidermal Growth Factor Receptor Positive Tumors.
Tumor-targeted near-infrared fluorescent dyes have the potential to improve cancer surgery by enabling surgeons to locate and resect more malignant lesions where good visualization tools are required to ensure complete removal of malignant tissue. Although the tumor-targeted fluorescent dyes used in humans to date have been either small organic molecules or high molecular weight antibodies, low molecular weight protein scaffolds have attracted significant attention because they penetrate solid tumors almost as efficiently as small molecules, but can be infinitely mutated to bind almost any antigen. Here we describe the use of a 10 kDa protein scaffold, a Centyrin, to target a near-infrared fluorescent dye to tumors that overexpress the epidermal growth factor receptor (EGFR) for fluorescence-guided surgery (FGS). We have developed and optimized the dose and time required for imaging small tumor burdens with minimal background fluorescence in real-time fluorescence-guided surgery of EGFR-expressing tumor xenografts in murine models. We demonstrate that the Centyrin-near-infrared dye conjugate (CNDC) binds selectively to human EGFR+ cancer cells with an EC50 of 2 nM, localizes to EGFR+ tumor xenografts in athymic nude mice and that uptake of the dye in xenografts is significantly reduced when EGFR are blocked by preinjection of excess unlabeled Centyrin. Taken together, these data suggest that CNDCs can be used for intraoperative identification and surgical removal of EGFR-expressing lesions and that Centyrins targeted to other tumor-specific antigens should prove similarly useful in fluorescence guided surgery of cancer. In addition, we demonstrate that the CNDC is detected in the NIR region of the spectrum and can be utilized for fluorescence-guided surgery (FGS). In addition, we propose that with its eventual complete clearance from EGFR-negative tissues and its quantitative retention in the tumor mass for >24 h, a Centyrin-targeted NIR dye should provide excellent tumor contrast when injected at least 6-8 h before initiation of cancer surgery in human patients.


Cognitive empathy contributes to poor social functioning in schizophrenia: Evidence from a new self-report measure of cognitive and affective empathy.
Cognitive empathy impairments have been linked to poor social functioning in schizophrenia. However, prior studies primarily used self-reported empathy measures developed decades ago that are not well-aligned with contemporary models of empathy. We evaluated empathy and its relationship to social functioning in schizophrenia using the recently developed Questionnaire of Cognitive and Affective Empathy (QCAE). Schizophrenia (n=52) and healthy comparison (n=37) subjects completed the QCAE, Interpersonal Reactivity Index (IRI), and measures of neurocognition, symptoms, and social functioning. Between-group differences on the QCAE, and relationships between QCAE and IRI subscales, neurocognition, symptoms, and social functioning were examined. The schizophrenia group reported significantly lower cognitive empathy than comparison subjects, which was driven by low online simulation scores. Cognitive empathy explained significant variance in social functioning after accounting for neurocognition and symptoms. Group differences for affective empathy were variable; the schizophrenia group reported similar proximal responsivity, but elevated emotion contagion relative to comparison subjects. These findings bolster support for the presence and functional significance of impaired cognitive empathy in schizophrenia using a contemporary measure of empathy. Emerging evidence that some aspects of affective empathy may be unimpaired or hyper-responsive in schizophrenia and implications for the assessment and treatment of empathy in schizophrenia are discussed.


A novel nuclease-ATPase (Nar71) from archaea is part of a proposed thermophilic DNA repair system.
We have identified a novel structure-specific nuclease in highly fractionated extracts of the thermophilic archaeon Methanothermobacter thermautotrophicus (Mth). The 71 kDa protein product of open reading frame mth1090 is a nuclease with ATPase activity, which we call Nar71 (Nuclease-ATPase in Repair, 71 kDa). The nar71 gene is located in a gene neighbourhood proposed by genomics to encode a novel DNA repair system conserved in thermophiles. The biochemical characterization of Nar71 presented here is the first analysis from within this neighbourhood, and it supports the insight from genomics. Nuclease activity of Nar71 is specific for 3' flaps and flayed duplexes, targeting single-stranded DNA (ssDNA) regions. This activity requires Mg2+ or Mn2+ and is greatly reduced in ATP. In ATP, Nar71 displaces ssDNA, also with high specificity for 3' flap and flayed duplex DNA. Strand displacement is weak compared with nuclease activity, but in ATPS it is abolished, suggesting that Nar71 couples ATP hydrolysis to DNA strand separation. ATPase assays confirmed that Nar71 is stimulated by ssDNA, though not double-stranded DNA. Mutation of Lys-117 in Nar71 abolished ATPase and nuclease activity, and we describe a separation-of-function mutant (K68A) that has lost ATPase activity but retains nuclease activity. A model of possible Nar71 function in DNA repair is presented.


Developmental changes in the metabolic network of snapdragon flowers.
Evolutionary and reproductive success of angiosperms, the most diverse group of land plants, relies on visual and olfactory cues for pollinator attraction. Previous work has focused on elucidating the developmental regulation of pathways leading to the formation of pollinator-attracting secondary metabolites such as scent compounds and flower pigments. However, to date little is known about how flowers control their entire metabolic network to achieve the highly regulated production of metabolites attracting pollinators. Integrative analysis of transcripts and metabolites in snapdragon sepals and petals over flower development performed in this study revealed a profound developmental remodeling of gene expression and metabolite profiles in petals, but not in sepals. Genes up-regulated during petal development were enriched in functions related to secondary metabolism, fatty acid catabolism, and amino acid transport, whereas down-regulated genes were enriched in processes involved in cell growth, cell wall formation, and fatty acid biosynthesis. The levels of transcripts and metabolites in pathways leading to scent formation were coordinately up-regulated during petal development, implying transcriptional induction of metabolic pathways preceding scent formation. Developmental gene expression patterns in the pathways involved in scent production were different from those of glycolysis and the pentose phosphate pathway, highlighting distinct developmental regulation of secondary metabolism and primary metabolic pathways feeding into it.


Basement membrane and fibroblast aberration in blisters at the donor, graft, and spontaneously healed sites in patients with burns.
Blisters that developed on spontaneously healed wounds and grafts in 13 patients with burns were analyzed by light, fluorescence, and electron microscopy. Blisters developed on the dermal side of the dermoepidermal junction and occurred more frequently in donor site and healed mesh graft than in split-thickness sheet graft. The four major components of the basement membrane zone (bullous pemphigoid antigen, laminin, type IV collagen, and epidermolysis bullosa acquisita antigen) were reduced in quantity and irregularly deposited at blister sites. Immediately adjacent to the blisters, epidermolysis bullosa acquisita antigen appeared normal in quantity, while laminin, type IV collagen, and bullous pemphigoid antigen levels appeared slightly reduced. Mononuclear infiltrates and autoantibodies were not detected by light microscopy or direct-indirect immunofluorescence, respectively. Ultrastructurally, adjacent dermal fibroblasts demonstrated swollen rough endoplasmic reticulum and vacuolization. We speculate that blister development in patients with burns is related to defective reorganization of the basement membrane zone in association with dermal fibroblast aberration during wound healing.


Public health impact and economic benefits of quadrivalent influenza vaccine in Latin America.
Annual trivalent influenza vaccines (TIV) containing 2 A strains and one B lineage have been recommended for the prevention of influenza in most of Latin American countries. However, the circulation of 2 B lineages (Victoria and Yamagata) and difficulties in predicting the predominating lineage have led to the development of quadrivalent influenza vaccines (QIV), including both B lineages. Thus, the objective was to estimate the public health impact and influenza-related costs if QIV would have been used instead of TIV in 3 Latin American countries. We used a static model over the seasons 2010-2014 in Brazil, 2007-2014 in Colombia and 2006-2014 in Panama, focusing on population groups targeted by local vaccination recommendations: young children, adults with risk factors and the elderly. In Brazil, between 2010 and 2014, using QIV instead of TIV would have avoided US$ 6,200 per 100,000 person-years in societal costs, based on 168 influenza cases, 89 consultations, 3.2 hospitalizations and 0.38 deaths per 100,000 person-years. In Colombia and Panama, these would have ranged from US$ 1,000 to 12,700 (based on 34 cases, 13-25 consultations, 0.6-8.9 hospitalizations and 0.04-1.74 deaths) and from US$ 3,000 to 33,700 (based on 113 cases, 55-82 consultations, 0.5-27.8 hospitalizations and 0.08-6.87 deaths) per 100,000 person-years, respectively. Overall, the broader protection offered by QIV would have reduced the influenza humanistic and economic burden in the 3 countries. Despite the lack of local data leading to several extrapolations, this study is the first to give quantitative estimates of the potential benefits of QIV in Latin America.


Association of Retinal Architecture, Intrathecal Immunity, and Clinical Course in Multiple Sclerosis.
Biomarkers to estimate long-term outcomes in patients with multiple sclerosis (MS) and to assign patients to individual treatment regimens are urgently needed. To assess whether retinal layer volumes are correlated with immune cell subsets and immunoglobulin indices in the cerebrospinal fluid and whether retinal layer volumes alone or in combination with intrathecal variables are associated with worsening of disease in patients with relapsing-remitting MS. This observational cohort study included 312 patients with relapsing-remitting MS in 2 independent cohorts (72 patients with short disease duration [cohort 1] and 240 patients with longer disease duration [cohort 2]) treated at a single German university hospital from April 15, 2013, through November 11, 2015. The common ganglion cell and inner plexiform layer (GCIPL) and inner nuclear layer (INL) volumes were tested for association with the immunoglobulin indices and the frequencies of immune cells in the cerebrospinal fluid (including B cells, T cells, and natural killer cells) (cohort 1). Volumes of GCIPL alone (cohorts 1 and 2) or GCIPL corrected for intrathecal B-cell frequencies (cohort 1) were tested for their association with worsening disability. A total of 312 patients (212 women [67.9%] and 100 men [32.1%]; median age, 34.0 years [interquartile range (IQR), 28.0-42.0 years]) were available for analysis. In cohort 1 (50 women [69.4%] and 22 men [30.6%]; median age, 31.0 years [IQR, 26.3-38.3 years]), with short disease durations (median, 1.0 months [IQR, 1.0-2.0 months]), low GCIPL volumes were associated with increased intrathecal B-cell frequencies (median, 1.96% [IQR, 1.45%-4.20%]) and intrathecal IgG synthesis (median cerebrospinal fluid/serum IgG index, 0.78 [IQR, 0.53-1.07]). The INL volumes correlated with the frequencies of intrathecal CD56bright natural killer cells (r = 0.28; P = .007). Individuals with low GCIPL volumes (<1.99 mm3) had a 6.4-fold risk for worsening disability during follow-up compared with patients with higher GCIPL values (95% CI, 1.7-24.2; P = .007). This finding was reproduced in cohort 2 (162 women [67.5%] and 78 men [32.5%]; median age, 34.0 years [IQR, 29.0-42.0 years]) consisting of patients with longer disease durations (median, 36.0 months [IQR, 21.0-60.0 months]) (hazard ratio, 2.4; 95% CI, 1.2-4.8; P = .02). In both cohorts, INL volumes correlated with the prospective increase in T2 lesion load and the number of gadolinium-enhancing lesions. Retinal layers reflect different aspects of disease activity during MS. Loss of GCIPL is associated with intrathecal B-cell immunity and constitutes an independent risk factor for worsening disability, whereas high INL volumes are associated with activity on magnetic resonance imaging in the brain parenchyma. Thus, retinal optical coherence tomography might be a means to support stratification of patients with MS for different therapeutic regimens.


The role of mitochondrial dynamics in cardiovascular diseases.
The process of mitochondrial dynamics is emerging as a core player in cardiovascular homeostasis. This process refers to the co-ordinated cycles of biogenesis, fusion, fission and degradation to which mitochondria constantly undergo to maintain their integrity, distribution and size. These mechanisms represent an early response to mitochondrial stress, confining organelle portions that are irreversibly damaged and preserving mitochondrial function. Accumulating evidence demonstrates that impairment in mitochondrial dynamics leads to myocardial damage and cardiac disease progression in a variety of disease models, including pressure overload, ischaemia/reperfusion and metabolic disturbance. These findings suggest that modulation of mitochondrial dynamics may be considered as a valid therapeutic strategy in cardiovascular diseases. In this review, we discuss the current evidence about the role of mitochondrial dynamics in cardiac pathophysiology, with a particular focus on the mechanisms underlying the development of cardiac hypertrophy and heart failure, metabolic and genetic cardiomyopathies, ischaemia/reperfusion injury, atherosclerosis and ischaemic stroke. LINKED ARTICLES: This article is part of a themed issue on Cellular metabolism and diseases. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v178.10/issuetoc.


Stimulation of apolipoprotein secretion in very-low-density and high-density lipoproteins from cultured rat hepatocytes by dexamethasone.
The effects of dexamethasone (a synthetic glucocorticoid) and insulin on the secretion of very-low-density lipoprotein (VLDL) and high-density lipoprotein (HDL) were investigated. Rat hepatocytes in monolayer culture were preincubated for 15 h in the presence or absence of combinations of 100 nM-dexamethasone and 2 nM-, 10 nM- or 50 nM-insulin. Dexamethasone increased [3H]oleate incorporation into secreted triacylglycerol by 2.7-fold and the mass of triacylglycerol secreted by 1.5-fold. Insulin alone decreased these parameters and antagonized the effect of dexamethasone. Dexamethasone increased the secretion of [3H]leucine in apolipoprotein (apo) E, and in the large (BH) and small (BI) forms of apo B in VLDL by about 7.1-, 3.6- and 4.0-fold respectively. Insulin alone decreased the secretion of these 3H-labelled apolipoproteins in VLDL. However, 2 nM-insulin with dexamethasone increased the secretion of 3H-labelled apo BH and apo BL by a further 0.8- and 3.2-fold respectively; 50 nM-insulin decreased the secretions of apo E, apo BH and apo BL in VLDL. Similar effects for dexamethasone or insulin alone were also obtained for the masses of apo E and apo BL + H secreted in VLDL. Albumin secretion was not significantly altered by either dexamethasone or insulin alone, but in combination they stimulated by 2.1-2.6-fold. Insulin or dexamethasone alone had little effect on the secretion of apolipoproteins in the HDL fraction. However, dexamethasone plus 2 nM-insulin increased the incorporation of [3H]leucine into apo AI, apo AH plus apo C, apo AIV and apo E of HDL by about 1.8-, 1.6-, 1.7- and 2.0-fold respectively. The apo E in the bottom fraction represented about 69% of the total 3H-labelled apo E secreted. The responses in the total secretion of apo E from the hepatocytes resembled those seen in HDL. The interactions of insulin and dexamethasone are discussed in relation to the general regulation of lipoprotein metabolism, the development of hyperlipidaemias and the predisposition to premature atherosclerosis.


Positron emission tomography/ultrasound fusion technique in patients with malignant melanoma.
(18)F-fluorodeoxyglucose positron emission tomography (FDG-PET)/CT is commonly used to assess tumour recurrence in high-risk patients with malignant melanoma (MM). However, results can be ambiguous either because of the CT's insufficient soft-tissue contrast or non-specific FDG accumulation caused by inflammation. Ultrasound (US) can provide additional morphologic information that is superior to CT. For precisely combining PET and US findings, we used a real-time fusion technique based on navigated US (PET/US fusion). Here, we describe our results from patients where PET/US fusion proved helpful in differentiating unclear PET/CT findings. This fusion technique is likely to be helpful for decision making in MM patients and biopsy guidance.


Testosterone Responders to Continuous Androgen Deprivation Therapy Show Considerable Variations in Testosterone Levels on Followup: Implications for Clinical Practice.
We determined whether men on continuous androgen deprivation therapy who achieve testosterone less than 0.7 nmol/l demonstrate subsequent testosterone elevations during followup and whether such events predict worse oncologic outcomes. We evaluated a random, retrospective sample of 514 patients with prostate cancer treated with continuous androgen deprivation therapy in whom serum testosterone was less than 0.7 nmol/l at University Health Network between 2007 and 2016. Patients were followed from the date of the first testosterone measurement of less than 0.7 nmol/l to progression to castrate resistance, death or study period end. Study outcomes were the development of testosterone elevations greater than 0.7, greater than 1.1 and greater than 1.7 nmol/l, and progression to a castrate resistant state. Survival curves were constructed to determine the rate of testosterone elevations. Multivariate Cox regression analysis was done to assess whether elevations predicted progression to castrate resistance. Median patient age was 74 years and median followup was 20.3 months. Within 5 years of followup 82%, 45% and 18% of patients had subsequent testosterone levels greater than 0.7, greater than 1.1 and greater than 1.7 nmol/l, respectively. In 96% to 100% of these patients levels less than 0.7 nmol/l were subsequently reestablished within 5 years. No patient baseline characteristic was associated with elevations and elevations were not a significant predictor of progression to a castrate resistant state. Men on continuous androgen deprivation therapy in whom initial testosterone is less than 0.7 nmol/l frequently show subsequent elevations in serum testosterone. Such a development should not trigger an immediate response from physicians as these events are prognostically insignificant with regard to oncologic outcomes. Levels are eventually reestablished at less than 0.7 nmol/l.


Functional roentgenometric evaluation of the cervical spine in the sagittal plane.
Neutral, flexed and extended lateral cervical radiographs were taken of sixteen male and fourteen female asymptomatic subjects and were evaluated in order to determine the amplitude of sagittal motion occurring intersegmentally. Using templates, the quantity of vertebral body displacement was expressed as a percentage ratio. Statistical analysis provided a radiographic definition of a normal range of intersegmental motion, hypomobility, articular fixation, hypermobility and instability. Other quantitative features of sagittal plane motion are discussed. Absolute hyper and hypomobility is defined by motion (% SBD) values falling between one and two standard deviations above normal (.60 to .72 and .38 to .26, respectively). Values greater than two standard deviations above normal were found to represent instability and functional blockage (greater than .72 and less than .26, respectively).


Expression of class 3 semaphorins and neuropilin receptors in the developing mouse tooth.
The semaphorins are a large family of secreted or cell-bound signals needed for the development of the nervous system. We compared mRNA expression of class 3 semaphorins (Sema3A, 3B, 3C and 3F) and their two receptors (Neuropilin-1 and -2) in the embryonic mouse first molar tooth germ (E10-18) by radioactive in situ hybridization. All genes showed distinct developmentally regulated expression patterns during tooth organogenesis. Interestingly, Sema3A and 3C were first detected in the early dental epithelium, and later both genes were present in the epithelial primary enamel knot, a putative signaling center of the embryonic tooth regulating tooth morphogenesis. Prior to birth, Sema3A was also observed in tooth-specific cells, preodontoblasts, which later differentiate into odontoblasts secreting dentin, and in the mesenchymal dental follicle cells surrounding the tooth germ. Sema3B appeared transiently in the dental mesenchyme in the bud and cap stage tooth while Sema3F was expressed in both epithelial and mesenchymal components of the tooth. Of note, Npn-1 expression pattern was largely complementary to that of Sema3A, and transcripts were restricted to the dental mesenchymal cells. Npn-1 expression was first seen in the developing dental follicle, and later transcripts also appeared in the dental papilla mesenchyme. In contrast, Npn-2 signal was seen in both epithelial and mesenchymal tissues such as in the primary enamel knot and preodontoblasts.


Complexation of oppositely charged polyelectrolytes: effect of discrete charge distribution along the chain.
Similar to the Debye-Hückel plasma, charged groups of oppositely charged polyelectrolytes attract each other. We find exact expression for the correlation free energy of electrostatic attraction in polyelectrolytes within the random phase approximation. The obtained expression is shown to be valid in dilute, semidilute, concentrated solutions, and melts. In this theory we explicitly take into account positions of charged groups on the chains and examine both charge and polymer concentrations fluctuations. The existence of the critical point in theta solvent is demonstrated. The theory reproduces scaling results and describes the system beyond the scaling approximation.


Correlates of participation in physical activity among older women in Israel: does ethno-cultural background matter?
Osteoporosis and its related fractures are major public health concerns. Physical activity (PA) is crucial for bone density preservation and fractures prevention. Yet, gaps in understanding exist regarding how ethno-cultural backgrounds might shape attitudes, intentions and actual PA participation. Based on the theory of planned behaviour (TPB) for predicting PA, the aims of this study were: (i) to compare attitudes, subjective norms, perceived control, intentions and knowledge, across four ethno-cultural groups; (ii) to evaluate the fit of the model we constructed across four ethno-cultural groups of women: Israeli-born Jews and Israeli-born Bedouin-Muslims, immigrants from the Former Soviet Union (FSU) and Ethiopian immigrants. Four hundred women (one hundred from each group), aged >65, completed valid and reliable questionnaires assessing knowledge, TPB components and actual PA. The level of knowledge on osteoporosis was relatively low among all four ethno-cultural groups. Intention to participate in PA was the only variable that directly predicted actual PA. Intention to participate in PA served as a mediator among attitudes, subjective norms, perceived control and actual PA. The structural equation models (SEMs) revealed that among Israeli-born Jews and Ethiopian immigrants, TPB components mediated the link between knowledge and intention to participate in PA. Among FSU immigrants and Israeli Bedouin-Muslims, the knowledge variable was not included in the final model, as its contribution was not significant. It is essential to better understand and augment interventions that enhance PA in the community, and to address the unique needs of each ethno-cultural group.


Gestational Exposures to Phthalates and Folic Acid, and Autistic Traits in Canadian Children.
The etiology of autism spectrum disorder is poorly understood. Few studies have investigated the link between endocrine-disrupting chemicals and autistic traits. We examined the relationship between gestational phthalates and autistic traits in 3- to 4-y-old Canadian children. We also investigated potential effect modification by sex and folic acid supplementation. We enrolled 2,001 women>18 years of age during the first trimester of pregnancy between 2008 and 2011 from 10 cities in Canada. At 3-4 years of age, 610 children underwent neuropsychological assessments including the Social Responsiveness Scale-II (SRS-2) as a measure of autistic traits and social impairment. We measured 11 phthalate metabolites in maternal first trimester urine samples and assessed folic acid supplementation from reported intakes. We estimated covariate-adjusted differences in SRS-2 T-scores with a doubling in phthalate concentrations in 510 children with complete data. Mean total SRS T-score was 45.3 (SD=6.1). Children with higher gestational exposure to mono-n-butyl (MBP) and mono-3-carboxypropyl (MCPP) concentrations exhibited significantly higher total SRS T-scores, indicating greater overall social impairment, as well as higher scores on subdomains, indicating deficits in social cognition, social communication, social motivation, and restricted interests/repetitive behaviors. A doubling in MBP or MCPP concentrations was associated with 0.6 (95% CI: 0.1, 1.0) and 0.5 (95% CI: 0.1, 0.8) higher total SRS T-scores. Associations were consistently and significantly stronger in boys (βMBP=1.0; 95% CI: 0.4, 1.6; n=252) compared with girls (βMBP=0.1; 95% CI: -0.6, 0.7; n=258) and among children who had lower prenatal folic acid supplementation (<400μg/d) (βMBP=1.3; 95% CI: 0.4, 2.3; n=59) compared with those who had adequate folic acid supplementation (≥400μg/d) (βMBP=0.4; 95% CI: -0.1, 0.8; n=451). Higher gestational concentrations of some phthalate metabolites were associated with higher scores of autistic traits as measured by the SRS-2 in boys, but not girls; these small size effects were mitigated by first trimester-of-pregnancy folic acid supplementation. https://doi.org/10.1289/EHP5621.


Bending strength of delaminated aerospace composites.
Buckling-driven delamination is considered among the most critical failure modes in composite laminates. This paper examines the propagation of delaminations in a beam under pure bending. A pre-developed analytical model to predict the critical buckling moment of a thin sub-laminate is extended to account for propagation prediction, using mixed-mode fracture analysis. Fractography analysis is performed to distinguish between mode I and mode II contributions to the final failure of specimens. Comparison between experimental results and analysis shows agreement to within 5 per cent in static propagation moment for two different materials. It is concluded that static fracture is almost entirely driven by mode II effects. This result was unexpected because it arises from a buckling mode that opens the delamination. For this reason, and because of the excellent repeatability of the experiments, the method of testing may be a promising means of establishing the critical value of mode II fracture toughness, G(IIC), of the material. Fatigue testing on similar samples showed that buckled delamination resulted in a fatigue threshold that was over 80 per cent lower than the static propagation moment. Such an outcome highlights the significance of predicting snap-buckling moment and subsequent propagation for design purposes.


Phoneme discrimination and mismatch negativity in English and Japanese speakers.
Neural templates for phonemes in one's native language are formed early in life; these can be modified but are difficult to form de novo. These can be examined with mismatch negativity (MMN). Three phonemic contrasts were presented to adult native English compared with Japanese speakers who acquired English later in life: vowels native to both languages (/i//iy/), consonant-vowel contrasts (/da//wa/) phonemic in both languages, and consonant-vowel contrasts phonemic in English but not in Japanese (/ra//la/). For vowels, no MMN differences were found. For /da//wa/, MMN amplitude was significantly reduced in Japanese speakers. For /ra//la/, only 50% of the Japanese group showed an identifiable MMN. This suggests that phonemic templates are formed early in life, and non-native consonant contrasts are difficult to learn later.


T-cell-replete haploidentical transplantation versus autologous stem cell transplantation in adult acute leukemia: a matched pair analysis.
Adult patients with acute leukemia in need of a transplant but without a genoidentical donor are usually considered upfront for transplantation with stem cells from any other allogeneic source, rather than autologous stem cell transplantation. We used data from the European Society for Blood and Marrow Transplantation and performed a matched pair analysis on 188 T-cell-replete haploidentical and 356 autologous transplants done from January 2007 to December 2012, using age, diagnosis, disease status, cytogenetics, and interval from diagnosis to transplant as matching factors. "Haploidentical expert" centers were defined as having reported more than five haploidentical transplants for acute leukemia (median value for the study period). The median follow-up was 28 months. Multivariate analyses, including type of transplant categorized into three classes ("haploidentical regular", "haploidentical expert" and autologous), conditioning intensity (reduced intensity versus myeloablative conditioning) and the random effect taking into account associations related to matching, showed that non-relapse mortality was higher following haploidentical transplants in expert (HR: 4.7; P=0.00004) and regular (HR: 8.98; P<10(-5)) centers. Relapse incidence for haploidentical transplants was lower in expert centers (HR:0.39; P=0.0003) but in regular centers was similar to that for autologous transplants. Leukemia-free survival and overall survival rates were higher following autologous transplantation than haploidentical transplants in regular centers (HR: 1.63; P=0.008 and HR: 2.31; P=0.0002 respectively) but similar to those following haploidentical transplants in expert centers. We conclude that autologous stem cell transplantation should presently be considered as a possible alternative to haploidentical transplantation in regular centers that have not developed a specific expert program.


Integrated bioinformatics approaches to investigate alterations in transcriptomic profiles of monkeypox infected human cell line model.
The recent re-emergence of the monkeypox (mpox) epidemic in nonendemic regions has raised concerns regarding a potential global outbreak. The mpox virus (MPV) is a smallpox-like virus belonging to the genus Orthopoxvirus (family: Poxviridae). Although studies suggest that MPV infection suppresses the Toll-like receptor-3- and tumor necrosis factor-α-related signaling pathways, whether MPV regulates other immune-related pathways remains unclear. In this study, two distinct temporal patterns were used for establishing an MPV-infected human immortal epithelial cancer cell line (HeLa). These two durations 2 and 12 h of incubation were selected to identify the coregulated genes and pathways affected by MPV infection. The use of the Gene Ontology framework, Kyoto Encyclopedia of Genes and Genome database, and MetaCore software yielded valuable insights. Specifically, various pathways were found to be enriched in HeLa cells infected with MPV for 2 and 12 h. These pathways included Notch, CD40, CD95, hypoxia-inducible factor-1-α, interleukin (IL)- 1, IL-6, phosphoinositide 3-kinase, nuclear factor-κB, mitogen-activated protein kinase, and oxidative stress-induced signalling pathways. Clusters and pathways of metabolism and viral replication cycles were significantly associated with the 2-hour infection group. This association was identified based on the regulation of genes such as HSPG2, RHPN2, MYL1, ASPHD2, CA9, VIPR1, SNX12, MGC2752, SLC25A1, PEX19, and AREG. Furthermore, clusters and pathways related to immunity and cell movement were found to be associated with the 12-hour infection group. This association was identified based on the regulation of genes such as C1orf21, C19orf48, HRK, IL8, GULP1, SCAND2, ATP5C1, FEZ1, SGSH, TACC2, CYP4X1, MMP1, CPB1, P2RY13, WDR27, PRPF4, and ENDOD1. This study can improve our understanding of the mechanisms underlying the pathophysiology and post-infection sequelae of mpox. Our findings provide valuable insights into the various modes of MPV infection.


Comparative proteomic analysis of biofilm and planktonic cells of Lactobacillus plantarum DB200.
This study investigated the relative abundance of extracellular and cell wall associated proteins (exoproteome), cytoplasmic proteins (proteome), and related phenotypic traits of Lactobacillus plantarum grown under planktonic and biofilm conditions. Lactobacillus plantarum DB200 was preliminarily selected due to its ability to form biofilms and to adhere to Caco2 cells. As shown by fluorescence microscope analysis, biofilm cells became longer and autoaggregated at higher levels than planktonic cells. The molar ratio between glucose consumed and lactate synthesised was markedly decreased under biofilm compared to planktonic conditions. DIGE analysis showed a differential exoproteome (115 protein spots) and proteome (44) between planktonic and biofilm L. plantarum DB200 cells. Proteins up- or downregulated by at least twofold (p < 0.05) were found to belong mainly to the following functional categories: cell wall and catabolic process, cell cycle and adhesion, transport, glycolysis and carbohydrate metabolism, exopolysaccharide metabolism, amino acid and protein metabolisms, fatty acid and lipid biosynthesis, purine and nucleotide metabolism, stress response, oxidation/reduction process, and energy metabolism. Many of the above proteins showed moonlighting behavior. In accordance with the high expression levels of stress proteins (e.g., DnaK, GroEL, ClpP, GroES, and catalase), biofilm cells demonstrated enhanced survival under conditions of environmental stress.


Immunolocalization of succinate dehydrogenase in the esophagus epithelium of domesticated mammals.
Using immunohistochemistry and transmission electron microscopy (TEM), the esophagus epithelia of seven domesticated mammals (horse, cattle, goat, pig, dog, laboratory rat, cat) of three nutrition groups (herbivorous, omnivorous, carnivorous) were studied to get first information about energy generation, as demonstrated by succinate dehydrogenase (SDH) activities. Distinct reaction intensities could be observed in all esophageal cell layers of the different species studied reflecting moderate to strong metabolic activities. The generally strong staining in the stratum basale indicated that new cells are continuously produced. The latter feature was confirmed by a thick, and in the horse generally highly active stratum spinosum. Only in the pig, reaction intensity variations occurred, obviously related to differences in physical feed quality or restricted feed allocation. The immunohistochemical results were corroborated by the presence of intact mitochondria in the esophageal cells of all species and nutrition types studied, except for the horse. Possible relationships between SDH reaction intensities and feed structure, mass or consistency are discussed.


Fast subplasma membrane Ca2+ transients control exo-endocytosis of synaptic-like microvesicles in astrocytes.
Astrocytes are the most abundant glial cell type in the brain. Although not apposite for long-range rapid electrical communication, astrocytes share with neurons the capacity of chemical signaling via Ca(2+)-dependent transmitter exocytosis. Despite this recent finding, little is known about the specific properties of regulated secretion and vesicle recycling in astrocytes. Important differences may exist with the neuronal exocytosis, starting from the fact that stimulus-secretion coupling in astrocytes is voltage independent, mediated by G-protein-coupled receptors and the release of Ca(2+) from internal stores. Elucidating the spatiotemporal properties of astrocytic exo-endocytosis is, therefore, of primary importance for understanding the mode of communication of these cells and their role in brain signaling. We here take advantage of fluorescent tools recently developed for studying recycling of glutamatergic vesicles at synapses (Voglmaier et al., 2006; Balaji and Ryan, 2007); we combine epifluorescence and total internal reflection fluorescence imaging to investigate with unprecedented temporal and spatial resolution, the stimulus-secretion coupling underlying exo-endocytosis of glutamatergic synaptic-like microvesicles (SLMVs) in astrocytes. Our main findings indicate that (1) exo-endocytosis in astrocytes proceeds with a time course on the millisecond time scale (tau(exocytosis) = 0.24 +/- 0.017 s; tau(endocytosis) = 0.26 +/- 0.03 s) and (2) exocytosis is controlled by local Ca(2+) microdomains. We identified submicrometer cytosolic compartments delimited by endoplasmic reticulum tubuli reaching beneath the plasma membrane and containing SLMVs at which fast (time-to-peak, approximately 50 ms) Ca(2+) events occurred in precise spatial-temporal correlation with exocytic fusion events. Overall, the above characteristics of transmitter exocytosis from astrocytes support a role of this process in fast synaptic modulation.


Foetal kidney maldevelopment in maternal use of angiotensin II type I receptor antagonists.
We report renal lesions observed in a foetus exposed throughout pregnancy to angiotensin II type I (AT 1) receptor antagonists. The mother suffered from essential hypertension and was treated with Cozaar (losartan 50 mg). Autopsy examination of the foetus revealed severe renal lesions, including tubular dysgenesis, hypertrophy of the endothelial and medial cells lining the arterial and arteriolar walls, hyperplasia of the juxtaglomerular apparatus and poorly developed vasa recta. Similar lesions have already been observed in foetuses of women treated with angiotensin-converting enzyme antagonists and also in foetuses and neonates of animals undergoing experimental blockade of the renin-angiotensin system. The purpose of this report is to describe structural lesions observed in the kidneys, and, particularly, vascular lesions. Our results suggest that the use of AT 1 receptor antagonists during pregnancy may have a severe deleterious effect on kidney development in the foetus.


Excess release of ferriheme in G6PD-deficient erythrocytes: possible cause of hemolysis and resistance to malaria.
Hemoglobin in glucose-6-phosphate dehydrogenase (G6PD)-deficient erythrocytes is abnormally vulnerable to oxidative denaturation, which may release ferriheme, a known cytolytic agent. We found 13.3 nmol of ferriheme in G6PD-deficient erythrocyte membranes (per gram of total erythrocyte hemoglobin) using a spectrophotometric assay, as compared to 9.8 in normal membranes (P less than .05). After incubation of erythrocytes with 250 mumol/L menadione, an oxidant drug, the values increased by 37.4 nmol in G6PD-deficient membranes and by 26 in normal membranes (P less than .005), indicating increased hemoglobin denaturation. To verify that hemoglobin denaturation in G6PD-deficient erythrocytes releases ferriheme in a form available to interact with other ligands, [14C]-chloroquine binding to intact erythrocytes was measured. With an initial concentration of 5 mumol/L chloroquine in a medium containing no menadione, an excess of 14.8 nmol of chloroquine was bound in G6PD-deficient erythrocytes (per gram of hemoglobin) as compared to normal erythrocytes (P less than .005). In the presence of 250 mumol/L menadione, chloroquine binding increased by 17.9 nmol in G6PD-deficient and by 7.2 in normal erythrocytes (P less than .005). These results indicate that ferriheme becomes available to interact with endogenous ligands and, thus, to mediate menadione-induced hemolysis in patients with G6PD deficiency. Furthermore, the increase in ferriheme may mediate the selective toxicity of menadione for Plasmodium falciparum parasites growing in G6PD-deficient erythrocytes. Ferriheme release in response to the intraerythrocytic oxidant stress introduced by malaria parasites also may account for the resistance to malaria afforded by G6PD deficiency. This is a US government work. There are no restrictions on its use.


Functional analysis of human microtubule-based motor proteins, the kinesins and dyneins, in mitosis/cytokinesis using RNA interference.
Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an Escherichia coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family), and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1, and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail.


Poly(L-lysine)-grafted-poly(ethylene glycol)-based surface-chemical gradients. Preparation, characterization, and first applications.
A simple dipping process has been used to prepare PEGylated surface gradients from the polycationic polymer poly(L-lysine), grafted with poly(ethylene glycol) (PLL-g-PEG), on metal oxide substrates, such as TiO(2) and Nb(2)O(5). PLL-g-PEG coverage gradients were prepared during an initial, controlled immersion and characterized with variable angle spectroscopic ellipsometry and x-ray photoelectron spectroscopy. Gradients with a linear change in thickness and coverage were generated by the use of an immersion program based on an exponential function. These single-component gradients were used to study the adsorption of proteins of different sizes and shapes, namely, albumin, immunoglobulin G, and fibrinogen. The authors have shown that the density and size of defects in the PLL-g-PEG adlayer determine the amount of protein that is adsorbed at a certain adlayer thickness. In a second step, single-component gradients of functionalized PLL-g-PEG were backfilled with nonfunctionalized PLL-g-PEG to generate two-component gradients containing functional groups, such as biotin, in a protein-resistant background. Such gradients were combined with a patterning technique to generate individually addressable spots on a gradient surface. The surfaces generated in this way show promise as a useful and versatile biochemical screening tool and could readily be incorporated into a method for studying the behavior of cells on functionalized surfaces.


Effects of endocrine disrupting chemicals in pigs.
Endocrine-disrupting chemicals (EDCs) are compounds that interfere with the expression, synthesis, and activity of hormones in organisms. They are released into the environment from flame retardants and products containing plasticizers. Persistent pesticides, such as dichlorodiphenyltrichloroethane (DDT) and hexachlorobenzene, also disrupt the endocrine system through interaction with hormone receptors. Endogenous hormones, such as 17β-estradiol (E2), are released in the urine and feces of farm animals and seep into terrestrial and aquatic ecosystems through sewage. Pigs are widely used as animal models to determine the effects of EDCs because they are physiologically, biochemically, and histologically similar to humans. EDCs primarily disrupt the reproductive and nervous systems of pigs. Moreover, embryonic development during the prenatal and early postnatal periods is particularly sensitive to EDCs. Mycotoxins, such as zearalenone, are food contaminants that alter hormonal activities in pigs. Mycotoxins also alter the innate immune system in pigs, making them vulnerable to diseases. It has been reported that farm animals are exposed to various types of EDCs, which accumulate in tissues, such as those of gonads, livers, and intestines. There is a lack of an integrated understanding of the impact of EDCs on porcine reproduction and development. Thus, this article aims to provide a comprehensive review of literature regarding the effects of EDCs in pigs.


Simulation of epiretinal prostheses - evaluation of geometrical factors affecting stimulation thresholds.
An accurate understanding of the electrical interaction between retinal prostheses and retinal tissue is important to design effective devices. Previous studies have used modelling approaches to simulate electric fields generated by epiretinal prostheses in saline and to simulate retinal ganglion cell (RGC) activation using passive or/and active biophysical models of the retina. These models have limited scope for studying an implanted human retinal prosthesis as they often do not account for real geometry and composition of the prosthesis-retina interface. This interface consists of real dimensions and location of stimulation and ground electrodes that are separated by the retinal tissue and surrounded by physiological fluids. In this study, we combined the prosthesis-retina interface elements into a framework to evaluate the geometrical factors affecting stimulation thresholds for epiretinal prostheses used in clinical human trials, as described by Balthasar et al. in their Investigative Ophthalmology and Visual Science (IOVS) paper published in 2008 using the Argus I epiretinal implants. Finite element method (FEM) based computations were used to estimate threshold currents based on a threshold criterion employing a passive electric model of the retina. Threshold currents and impedances were estimated for different electrode-retina distances. The profiles and the values for thresholds and impedances obtained from our simulation framework are within the range of measured values in the only elaborate published clinical trial until now using Argus I epiretinal implants. An estimation of resolution for the electrodes used in these trials was provided. Our results reiterate the importance of close proximity between electrodes and retina for safe and efficient retinal stimulation. The validation of our simulation framework being relevant for epiretinal prosthesis research is derived from the good agreement of the computed trends and values of the current study with measurements demonstrated in existing clinical trials on humans (Argus I). The proposed simulation framework could be used to generate the relationship between threshold and impedance for any electrode geometry and consequently be an effective tool for design engineers, surgeons and electrophysiologists.


Relationships between followers' behaviors and job satisfaction in a sample of nurses.
The study investigated two followership behaviors, followers' active engagement and followers' independent critical thinking, and their relationship with job satisfaction in a sample of nurses. In addition, the study also considered a number of control variables and classical job demands and job resources-workload and emotional dissonance for job demands, and meaningful work for job resources-which have an impact on well-being at work. A paper-and-pencil questionnaire was administered to 425 nurses in an Italian hospital, and a hierarchical multiple regression was used to test the hypotheses. In addition to the job demands and job resources considered, followers' active engagement had a significant impact on job satisfaction. Moreover, it showed a significant linear and curvilinear relationship with the outcome variable. Followers' independent critical thinking has a non significant relationship with job satisfaction, confirming the mixed results obtained in the past for this dimension. These findings bore out the importance of analyzing followers' behaviors as potential resources that people can use on the job to increase their own well-being. Looking at followers not just as passive recipients but as active and proactive employees can also benefit the organization.


Health benefits of a reduction of PM10 and NO2 exposure after implementing a clean air plan in the Agglomeration Lausanne-Morges.
Exposure to urban air pollution has been associated with adverse effects on cardio-vascular and respiratory health, both short and long term. Consequently, governments have applied policies to reduce air pollution. Quantitative health impact assessments of hypothetic changes in air pollution have been conducted at national and global level, but assessments of observed air pollution changes associated with specific clean air policies at a local or regional scale remain scarce. This study estimates health impacts attributable to a decrease in PM10 and NO2 exposure in the Agglomeration of Lausanne-Morges (ALM), Switzerland, between 2005 and 2015, corresponding to the implementation period of a supra-municipal plan of measures to reduce air pollution in different sectors such as transport, energy, and industry (called Plan OPair 05). The health impact assessment compares health effects attributed to air pollution exposure levels in 2015 (reference case) with those in 2005 (counterfactual scenario), using 2015 as baseline for all other input data. In the ALM, the modeled PM10 exposure reduction of 3.3μg/m3 from 2005 to 2015 prevents 26 premature deaths (equivalent to around 290 years of life lost), 215 hospitalization days due to cardio-vascular and respiratory diseases as well as approximately 47,000 restricted activity days annually. Monetized health impacts of the reduction of PM10 exposure are valued at approximately CHF 36 million annually. Immaterial costs, mainly related to the economic valuation of years of life lost, dominate the monetized health impacts (90% of total value), while savings at the workplace (net loss in production and reoccupation costs) amount to about CHF 1.9 million, and savings in health care costs to about CHF 0.5 million. The assessment is sensitive to the value assigned to immaterial costs and to uncertainties in the relative risk estimates, whereas variations in the baseline year (i.e. using 2005 data instead of 2015 data) affect results to a much lower degree. The alternative calculation based on NO2 exposure, which dropped by 5.6μg/m3, suggests the prevention of 51 premature deaths (equivalent to around 550 years of life lost) overall impacts valued at CHF 49 million. All in all, the reduction in mortality due to the air quality improvements accounts for (depending on the considered pollutant) about 1% to 2% of total all-cause annual mortality in the ALM population or 4-8 times larger than the annual traffic fatalities in the ALM.


FGF-8 is associated with anteroposterior patterning and limb regeneration in Xenopus.
FGF-8 has attracted attention particularly because of its importance for limb development in the chick and mouse, although it also has a number of earlier expression domains in these species. We have now cloned an FGF-8 homologue from Xenopus in which it is easier to do functional studies on early development. There is no maternal expression, while zygotic expression is highest in the gastrula and neurula stages. XFGF-8 is expressed as a ring around the blastopore and subsequently in the tail bud. There are several domains in the head including the hatching gland, the branchial clefts, and the midbrain-hindbrain border. At later stages there is a prominent band of expression in the limb bud epidermis. Although there is no morphological apical ridge, this band of expression suggests that the Xenopus limb bud contains a cryptic region with a similar ability to stimulate mesenchymal outgrowth. The mesoderm-inducing activity of XFGF-8 is somewhat lower than that of other FGFs, while the posteriorizing activity is similar. These differences are probably due to the different receptor specificity. The posterior expression and high posteriorizing activity suggest that XFGF-8 contributes to the patterning of the anterior-posterior axis by FGF family members during gastrulation. In contrast to the amniotes, Xenopus limb buds can regenerate following damage. We show that regeneration is correlated with the reexpression of XFGF-8 in the distal epidermis, suggesting that this ability is critical for successful limb regeneration.


The status of malaria among pregnant women: a study in Lagos, Nigeria.
A study was carried out on the status of malaria among 800 randomly selected pregnant women in Lagos State, Nigeria. Blood samples were obtained from finger pricking and tested for malaria parasites in thin blood films and 60% prevalence of malaria parasites was obtained. Interviews were conducted and structured questionnaires were administered to the pregnant women to obtain information on the clinical and social aspects of malaria. Results show that primigravidae accounted for a greater part of the 60% prevalence of malaria that affected mainly women in their 1st to 3rd month of pregnancy. The ages of the infected women ranged from 30 to 39 years (77%). Women with blood groups A and O had the highest prevalence of malaria, but there was no statistically significant difference between them and the uninfected women. Women with genotype AA had the highest prevalence of malaria, while pregnant women in Ikeja division had the highest incidence of malaria (41.7%). Majority of the infected women believed that mosquito bites and stress were responsible for their infection. Only 21.8% of the women did not associate mosquitoes with malaria. All the women were familiar with the symptoms of malaria but did not see it as a serious disease that could lead to death. Most of the women used bed nets but not the impregnated brands. There is need to educate women, especially during antenatal visits, on the severity of malaria and the risk of their susceptibility to it during pregnancy.


Bone and nutrition in elderly women: protein, energy, and calcium as main determinants of bone mineral density.
Nutrition is an important factor in the prevention and treatment of osteoporosis. Our goal was to examine the relationship between various nutrients and bone mass of several skeletal sites in elderly women, taking into account possible confounding variables. A cross-sectional study in 136 healthy Caucasian, postmenopausal women, free of medications known to affect bone was carried out. Bone mineral density (BMD) and body composition (lean and fat tissue) were measured by dual X-ray absorptiometry using specialized software for different skeletal sites. Parathyroid hormone (PTH) and vitamin D, 25(OH)D, as possible confounders, were determined in serum samples. Dietary intake, including all supplements, was assessed by 3-day dietary record and analyzed using Food Processor. Past physical activity and present walking were examined as well and accounted for as potential confounders. Simple and multiple regression models were created to assess the relationships between nutrients and BMD. To examine the co-linear variables and their possible independent association with bone, subgroup analyses were performed. : Showed independent influence of calcium, energy, and protein, examined separately and in multiple regression models on BMD of several skeletal sites. Magnesium, zinc and vitamin C were significantly related to BMD of several skeletal sites in multiple regression models (controlled for age, fat and lean tissue, physical activity and energy intake), each contributing more than 1% of variance. Serum PTH and 25(OH)D did not show significant association with bone mass. Despite the cross-sectional nature of our study we were able to show a significant relationship between BMD and several critical nutrients: energy, protein, calcium, magnesium, zinc and vitamin C. The exact involvement of these nutrients and their clinical significance in bone health need to be further elucidated in humans and conclusions about the effects of a single nutrient on bone mass must be given cautiously, taking into account its interaction and co-linearity with others. Understanding relationships among nutrients, not just limited to calcium and vitamin D, but others that have not been investigated to such extent, is an important step toward identifying preventive measures for bone loss and prevention of osteoporosis.


Hydroxyl radical generation by electron paramagnetic resonance as a new method to monitor ambient particulate matter composition.
Epidemiological studies have demonstrated the relationship between exposure to ambient particulate matter (PM) and health effects in those with cardiopulmonary diseases. The free radical generating activity of particles has been suggested as a unifying factor in the biological activity of PM in toxicological studies but so far has not been applied as a method for environmental monitoring of PM. The purpose of this study was to characterize hydroxyl radical (OH*) production by different size fractions of PM, to use as an alternative method for monitoring of PM composition and activity. We have developed a method, using electron paramagnetic resonance (EPR), to measure OH* radical formation in suspensions of particles in the presence of hydrogen peroxide and 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a specific spin-trap. Samples of ambient particulate matter (PM) of different size fractions were collected from various sites on various filters. PM deposited on filters as well as suspensions in water retain its ability to generate OH* and this generation is determined by concentration of hydrogen peroxide and soluble metals. However, large variations in OH* radical formation and kinetics were found with different soluble metals and within metals (Fe, V) with different valencies. The method was applied to environmental monitoring in Hettstedt-Zerbst, situated in South-Eastern Germany, where it showed a relation to Cu-content of PM. The method was also applied in Duisburg, where the PMI fraction showed the highest DMPO-OH* generation but was not linked to particle counts. The method integrates metal bioavailability and reactivity and can provide a better understanding of the effect of small variations in mass concentrations on health.


Intra-abdominal hypertension and the abdominal compartment syndrome.
The pressure within the abdominal cavity is normally little more than atmospheric pressure. However, even small increases in intra-abdominal pressure can have adverse effects on renal function, cardiac output, hepatic blood flow, respiratory mechanics, splanchnic perfusion and intracranial pressure. Although intra-abdominal pressure can be measured directly, this is invasive and bedside measurement of intra-abdominal pressure is usually achieved via the urinary bladder. This cheap, easy approach has been shown to produce results that correlate closely with directly measured abdominal pressures. Significant increases in intra-abdominal pressure are seen in a wide variety of conditions commonly encountered in the intensive care unit, such as ruptured aortic aneurysm, abdominal trauma and acute pancreatitis. Abdominal compartment syndrome describes the combination of increased intra-abdominal pressure and end-organ dysfunction. This syndrome has a high mortality, most deaths resulting from sepsis and multi-organ failure. Detection of abdominal compartment syndrome requires close surveillance of intra-abdominal pressure in patients thought to be at risk of developing intra-abdominal hypertension. The only available treatment for established abdominal compartment syndrome is decompressive laparotomy. Prevention of abdominal compartment syndrome after laparotomy by adoption of an open abdomen approach may be preferable in the patient at significant risk of developing intra-abdominal hypertension, but this has not been demonstrated in any large trials. Most surgeons prefer to adopt a 'wait and see' policy, only intervening when clinical deterioration is associated with a significant increase in intra-abdominal pressure.


Influence of staff behavior on infectious risk in operating rooms: what is the evidence?
SUMMARY A systematic literature review was performed to assess the impact of surgical-staff behaviors on the risk of surgical site infections. Published data are limited, heterogeneous, and weakened by several methodological flaws, underlying the need for more studies with accurate tools. OBJECTIVE To assess the current literature regarding the impact of surgical-staff behaviors on the risk of surgical-site infection (SSI). DESIGN Systematic literature review. METHODS We searched the Medline, EMBASE, Ovid, Web of Science, and Cochrane databases for original articles about the impact of intraoperative behaviors on the risk of SSI published in English before September 2013. RESULTS We retrieved 27 original articles reporting data on number of people in the operating room (n=14), door openings (n=14; number [n=6], frequency [n=7], reasons [n=4], or duration [n=3]), surgical-team discipline (evidence of distraction; n=4), compliance with traffic measures (n=6), or simulated behaviors (n=3). Most (59%) articles were published in 2009-2013. End points were the 30-day SSI rate (n=8), air-particle count (n=2), or microbiological air counts (n=6); 11 studies were only descriptive. Number of people in the operating room and SSI rate or airborne contaminants (particle/bacteria) were correlated in 2 studies. Door openings and airborne bacteria counts were correlated in 2 observational studies and 1 experimental study. Two cohort studies showed a significant association between surgeon interruptions/distraction or noise and SSI rate. The level of evidence was low in all studies. CONCLUSIONS Published data about the impact of operating-room behaviors on the risk of infection are limited and heterogeneous. All studies exhibit major methodological flaws. More studies with accurate tools should be performed to address the influence of operating room behaviors on the infectious risk.


An improved approach for the synthesis of alpha,alpha-dialkyl glycine derivatives by the Ugi-Passerini reaction.
A general and simple strategy for routine peptide synthesis with alpha,alpha-dialkyl glycines taking advantage of the four-component Ugi-Passerini reaction is presented. The isonitrile required for the reaction can be relatively simple and its selection based on cost, as the group it generates is easily removed under acidic conditions: in addition, this removal is not visibly affected by the bulkiness of the alpha-alkyl groups. Being a good leaving group from the N-terminal amino group of the amino acid, 4-methoxybenzyl was the choice for the amine component of the reaction. The method is illustrated with the synthesis of a series of acyl derivatives of several alpha,alpha-dialkyl glycines. The preparation of the latter compounds is also reported.


Extracellular matrix in the trabecular meshwork.
The extracellular matrix (ECM) of the trabecular meshwork (TM) is thought to be important in regulating intraocular pressure (IOP) in both normal and glaucomatous eyes. IOP is regulated primarily by a fluid resistance to aqueous humor outflow. However, neither the exact site nor the identity of the normal resistance to aqueous humor outflow has been established. Whether the site and nature of the increased outflow resistance, which is associated with open-angle glaucoma, is the same or different from the normal resistance is also unclear. The ECMs of the TM beams, juxtacanalicular region (JCT) and Schlemm's canal (SC) inner wall are comprised of fibrillar and non-fibrillar collagens, elastin-containing microfibrils, matricellular and structural organizing proteins, glycosaminoglycans (GAGs) and proteoglycans. Both basement membranes and stromal ECM are present in the TM beams and JCT region. Cell adhesion proteins, cell surface ECM receptors and associated binding proteins are also present in the beams, JCT and SC inner wall region. The outflow pathway ECM is relatively dynamic, undergoing constant turnover and remodeling. Regulated changes in enzymes responsible for ECM degradation and biosynthetic replacement are observed. IOP homeostasis, triggered by pressure changes or mechanical stretching of the TM, appears to involve ECM turnover. Several cytokines, growth factors and drugs, which affect the outflow resistance, change ECM component expression, mRNA alternative splicing, cellular cytoskeletal organization or all of these. Changes in ECM associated with open-angle glaucoma have been identified.