Quantitative Point of Care Tests for Timely Diagnosis of Early-Onset Preeclampsia with High Sensitivity and Specificity.
Preeclampsia is a heterogeneous and multiorgan cardiovascular disorder of pregnancy. Here, we report the development of a novel strip-based lateral flow assay (LFA) using lanthanide-doped upconversion nanoparticles conjugated to antibodies targeting two different biomarkers for detection of preeclampsia. We first measured circulating plasma FKBPL and CD44 protein concentrations from individuals with early-onset preeclampsia (EOPE), using ELISA. We confirmed that the CD44/FKBPL ratio is reduced in EOPE with a good diagnostic potential. Using our rapid LFA prototypes, we achieved an improved lower limit of detection: 10 pg ml-1 for FKBPL and 15 pg ml-1 for CD44, which is more than one order lower than the standard ELISA method. Using clinical samples, a cut-off value of 1.24 for CD44/FKBPL ratio provided positive predictive value of 100 % and the negative predictive value of 91 %. Our LFA shows promise as a rapid and highly sensitive point-of-care test for preeclampsia.


Acaricides containing zein nanoparticles: A tool for a lower impact control of the cattle tick Rhipicephalus microplus.
Nanoformulations containing zein nanoparticles (ZN) can promote the stability and protection of molecules with acaricidal activity. The present study sought to develop nanoformulations with ZN associated with cypermethrin (CYPE) + chlorpyrifos (CHLO) + a plant compound (citral, menthol or limonene), characterize them, and verify their efficacy against Rhipicephalus microplus ticks. Additionally, we aimed to assess its safety in nontarget nematodes found in soil at a site subjected to contamination by acaricides. The nanoformulations were characterized by dynamic light scattering and nanoparticle tracking analysis. Nanoformulations 1 (ZN+CYPE+CHLO+citral), 2 (ZN+CYPE+CHLO+menthol), and 3 (ZN+CYPE+CHLO+limonene) were measured for diameter, polydispersion, zeta potential, concentration, and encapsulation efficiency. Nanoformulations 1, 2, and 3 were evaluated in a range from 0.004 to 0.466 mg/mL on R. microplus larvae and caused mortality > 80% at concentrations above 0.029 mg/mL. The commercial acaricide Colosso® (CYPE 15 g + CHLO 25 g + citronellal 1 g) was evaluated also from 0.004 to 0.512 mg/mL and resulted in 71.9% larval mortality at 0.064 mg/mL. Formulations 1, 2, and 3 at 0.466 mg/mL showed acaricidal efficacy of 50.2%, 40.5%, and 60.1% on engorged females, respectively, while Colosso® at 0.512 mg/mL obtained only 39.4%. The nanoformulations exhibited long residual period of activity and lower toxicity to nontarget nematodes. ZN was able to protect the active compounds against degradation during the storage period. Thus, ZN can be an alternative for the development of new acaricidal formulations using lower concentrations of active compounds.


Cav2.2-NFAT2-USP43 axis promotes invadopodia formation and breast cancer metastasis through cortactin stabilization.
Distant metastasis is the main cause of mortality in breast cancer patients. Using the breast cancer genomic data from The Cancer Genome Atlas (TCGA), we identified brain specific Cav2.2 as a critical regulator of metastasis. Cav2.2 expression is significantly upregulated in breast cancer and its higher expression is inversely correlated with survival suggesting a previously unappreciated role of Cav2.2 in breast cancer. Cav2.2 is required for breast cancer migration, invasion, and metastasis. Interestingly, Cav2.2 promotes invadopodia formation and extracellular matrix (ECM) degradation through the stabilization of invadopodia component cortactin in a proteosome-dependent manner. Moreover, deubiquitinating enzyme USP43 mediated the functions of Cav2.2 in cortactin stabilization, invadopodia formation, ECM degradation, and metastasis. Interestingly, Cav2.2 upregulates USP43 expression through NFAT2 dephosphorylation and nuclear localization. Our study uncovered a novel pathway that regulates cortactin expression and invadopodia formation in breast cancer metastasis.


Metabolism and nutrition in patients with moderate and severe traumatic brain injury: A systematic review.
To examine the evidence on the metabolic state and nutritional treatment of patients with moderate-to-severe traumatic brain injury (TBI). A systematic review of the literature. From 1547 citations, 232 articles were identified and retrieved for text screening. Thirty-six studies fulfilled the criteria and 30 were accepted for data extraction. Variations in measurement methods and definitions of metabolic abnormalities hampered comparison of studies. However, consistent data demonstrated increased metabolic rate (96-160% of the predicted values), of hypercatabolism (-3 to -16 g N per day) and of upper gastrointestinal intolerance in the majority of the patients during the first 2 weeks after injury. Data also indicated a tendency towards less morbidity and mortality in early fed patients. The impact of timing, content and ways of administration of nutritional support on neurological outcome after TBI remains to be demonstrated.


Serum IL-36 cytokines levels in idiopathic pulmonary fibrosis and connective tissue disease-associated interstitial lung diseases.
Interleukin-36 (IL-36) family is associated with several fibrosis-related disorders and connective tissue diseases. However, their expression in idiopathic pulmonary fibrosis (IPF) and connective tissue disease-interstitial lung disease (CTD-ILD) is unknown. We included 19 CTD-ILD patients, 16 IPF patients, and 27 healthy control subjects. Determination of serum concentrations of IL-36α, IL-36γ and IL-36 receptor antagonist (IL-36Ra) was performed by ELISA. The value of biomarkers for the diagnosis and assessment of ILD was assessed by lung function tests and high-resolution computed tomography. Serum concentrations of IL-36α and IL-36γ in patients with CTD-ILD and IPF were significantly higher than that in healthy controls, whereas serum IL-36Ra concentrations were not significantly different between the 3 groups. Increased IL-36 levels correlated with disease severity in IPF patients. ROC curve analysis showed that the AUC was 0.9931 for IL-36α and 0.8194 for IL-36γ in IPF group. In CTD-ILD group, the AUC was 0.9825 for IL-36α and 0.7973 for IL-36γ. We demonstrated an imbalance in the agonist and antagonist profiles of IL-36 cytokines in ILD. IL-36 cytokines may be a new diagnostic or therapeutic target in ILD, especially in IPF.


Rescue of Rous sarcoma virus (RSV) from RSV-transformed human embryonic cells by cell fusion with chick embryo fibroblasts using lysolecithin.
Heterokaryon formation and Rous sarcoma virus (RSV)-induction were studied by fusion of RSV-transformed human embryonic cells with chick embryo fibroblasts in the presence of lysolecithin. Heterokaryon formation was observed by autoradiography. RSV-induction was identified by focus formation, electron microscopy and density gradient centrifugation of 3H-uridine-labeled particles. The most effective concentration of lysolecithin for virus induction was 10 mug/10(6) cells/0.1 ml. Efficiency of lysolecithin in virus induction was not less than that of ultraviolet-inactivated Sendai virus (UV-HVJ).


Location of cardiac arrest and impact of pre-arrest chronic disease and medication use on survival.
Cardiac arrest in a private location is associated with a higher mortality when compared to public location. Past studies have not accounted for pre-arrest factors such as chronic disease and medication. To investigate whether the association between cardiac arrest in a private location and a higher mortality can be explained by differences in chronic diseases and medication. We identified 27,771 out-of-hospital cardiac arrest patients ≥18 years old from the Danish Cardiac Arrest Registry (2001-2012). Using National Registries, we identified pre-arrest chronic disease and medication. To investigate the importance of cardiac arrest related factors and chronic disease and medication use we performed adjusted Cox regression analyses during day 0-7 and day 8-365 following cardiac arrest to calculate hazard ratios (HR) for death. Day 0-7: Un-adjusted HR for death day 0-7 was 1.21 (95%CI:1.18-1.25) in private compared to public location. When including cardiac arrest related factors HR for death was 1.09 (95%CI:1.06-1.12). Adding chronic disease and medication to the analysis changed HR for death to 1.08 (95%CI:1.05-1.12). 8-365 day: The un-adjusted HR for death day 8-365 was 1.70 (95% CI: 1.43-2.02) in private compared to public location. When including cardiac arrest related factors the HR decreased to 1.39 (95% CI: 1.14-1.68). Adding chronic disease and medication to the analysis changed HR for death to 1.27 (95% CI:1.04-1.54). The higher mortality following cardiac arrest in a private location is partly explained by a higher prevalence of chronic disease and medication use in patients surviving until day 8.


Erratum: Building effective machine learning models for ankle joint power estimation during walking using FMG sensors.
[This corrects the article DOI: 10.3389/fnbot.2022.836779.].


Adult survival in migratory caribou is negatively associated with MHC functional diversity.
Genes of the major histocompatibility complex (MHC) are involved in acquired immunity in vertebrates. Only a few studies have investigated the fitness consequences of MHC gene diversity in wild populations. Here, we looked at the association between annual survival and body mass and MHC-DRB exon 2 (MHC-DRB) genetic diversity, obtained from high-throughput sequencing, in two declining migratory caribou (Rangifer tarandus) herds. To disentangle the potential direct and general effects of MHC-DRB genetic diversity, we compared different indices of diversity that were either based on DNA-sequence variation or on physicochemical divergence of the translated peptides, thereby covering a gradient of allelic-to-functional diversity. We found that (1) body mass was not related to MHC-DRB diversity or genotype, and (2) adult survival probability was negatively associated with point accepted mutation distance, a corrected distance that considers the likelihood of each amino acid substitution to be accepted by natural selection. In addition, we found no evidence of fluctuating selection over time on MHC-DRB diversity. We concluded that direct effects were involved in the negative relationship between MHC functional diversity and survival, although the mechanism underlying this result remains unclear. A possible explanation could be that individuals with higher MHC diversity suffer higher costs of immunity (immunopathology). Our results suggest that genetic diversity is not always beneficial even in genes that are likely to be strongly shaped by balancing selection.


Construction of Biofunctionalized Anisotropic Hydrogel Micropatterns and Their Effect on Schwann Cell Behavior in Peripheral Nerve Regeneration.
Hydrogels have promising application in tissue regeneration due to their excellent physicochemical and biocompatible properties, whereas anisotropic micropatterns are been proven to directionally induce cell alignment and accelerate cell migration. However, an effect of biofunctionalized anisotropic hydrogel micropatterns on nerve regeneration has rarely been reported. In this study, the anisotropic polyacrylamide (PAM) hydrogel micropatterns with aligned ridge/groove structures were first prepared via in situ free radical polymerization and micromolding, and then biofunctionalized using YIGSR peptide for better promoting cell growth. The morphology, swelling ratio, wettability, mechanical properties, and stability of the prepared hydrogel were characterized. The successful immobilization of YIGSR peptide on the PAM hydrogel was monitored using FTIR, immunofluorescence staining, and ELISA. The effects on adhesion, directional growth, and biological function of Schwann cells were evaluated. The results displayed that the anisotropic PAM hydrogel micropatterns with inner porous structure possessed good stability, swelling, and mechanical properties. The YIGSR peptide could be well immobilized on hydrogel micropatterns with a percentage of 62.6%. The biofunctionalized anisotropic hydrogel micropatterns could effectively regulate the orientation growth of Schwann cells, and obviously up-regulate BDNF (40%) and β-actin (50%) expression compared with single hydrogel micropatterns, without negatively affecting the normal secretion of neurotropic factors by Schwann cells. To the best of our knowledge, this is the first time to study the construction and effect of biofunctionalized anisotropic hydrogel micropatterns on nerve regeneration, which may provide an experimental and theoretical basis for the design and development of artificial implants for nerve regeneration application.


Extracellular secretion of a cutinase with polyester-degrading potential by E. coli using a novel signal peptide from Amycolatopsis mediterranei.
Recent studies in this laboratory showed that an extracellular cutinase from A. mediterranei (AmCut) was able to degrade the plastics polycaprolactone and polybutylene succinate. Such plastics can be slow to degrade in soils due to a lack of efficient polyester degrading organisms. AmCut also showed potential for the biocatalytic synthesis of esters by reverse hydrolysis. The gene for AmCut has an upstream leader sequence whose transcript is not present in the purified enzyme. In this study, we show using predictive modelling, that this sequence codes for an N-terminal signal peptide that directs transmembrane expression via the Sec secretion pathway. E. coli is a useful host for recombinant enzymes used in biocatalysis due to the ease of genetic manipulation in this organism, which allows tuning of enzymes for specific applications, by mutagenesis. When a truncated GST-tagged AmCut gene (lacking its signal peptide) was expressed in E. coli, all cutinase activity was observed in the cytosolic fraction. However, when GST-tagged AmCut was expressed in E. coli along with its native signal peptide, cutinase activity was observed in both the periplasmic space and the culture medium. This finding revealed that the native signal peptide of a Gram-positive organism (AmCut) was being recognised by the Gram-negative (E. coli) Sec transmembrane transport system. AmCut was transported into E. coli's periplasmic space from where it was released into the culture medium. Surprisingly, the presence of a bulky GST tag at the N-terminus of the signal peptide did not hinder transmembrane targeting. Although the periplasmic targeting was unexpected, it is not unprecedented due to the conservation of the Sec pathway across species. It was more surprising that AmCut was secreted from the periplasmic space into the culture medium. This suggests that extracellular AmCut translocation across the E. coli outer membrane may involve non-classical secretion pathways. This tuneable recombinant E. coli expressing extracellular AmCut may be useful for degradation of polyester substrates in the environment; this and other applications are discussed.


Infrapopliteal bypass for severe ischemia: comparison of autogenous vein, composite, and prosthetic grafts.
Results of 253 consecutive bypass grafts to infrapopliteal arteries were reviewed. Most (92%) were placed for rest pain (103) or tissue loss (130). Autogenous veins were used in 175 (69%) cases, composite vein-prosthetic grafts were used in 45 (18%), and prosthetic grafts alone were used in 33 (13%). Follow-up ranged from 0 to 101 months (mean, 19 months); 37 grafts (15%) were lost to follow-up. The operative mortality rate was 4%, and 5-year patient survival rate was 44%. Limb salvage was 82% at 5 years. The 5-year patency of vein grafts (63%) exceeded that of both composite (28%) and prosthetic (7%) grafts (p = 0.005 and p = 0.00007, respectively); but the patency of composite and prosthetic grafts did not differ significantly (p = 0.29). The patency of reversed vein (59%) and in situ vein grafts (74%) was not significantly different at 5 years (p = 0.34). Patency was also not affected by the site of the proximal or distal anastomoses or diabetes. The major determinant of long-term patency in infrapopliteal reconstructions continues to be graft material. Composite grafts offered no clear advantage over prosthetic grafts, and both should be used only when there is no other alternative to amputation.


Cluster headache and obstructive sleep apnea: are they related disorders?
Patients with cluster headache (CH) have a higher prevalence of sleep apnea, and a possible relationship between these two conditions has been proposed. Although patients suffering from CH attacks often wake up from sleep, sleep apnea has been suggested to be a trigger or an associated abnormality in CH. It has been proposed that regulation of the hypothalamus may be responsible for sleep apnea, and that similiarly CH is generated in the hypothalamus. However, there is evidence that CH and obstructive sleep apnea are not causal, but rather parallel processes both generated in the hypothalamus. The exact role that sleep apnea plays in the perpetuation or precipitation of CH is still to be determined. This paper discusses the proposed pathophysiological mechanisms of these two entities and the possible relationship between CH and sleep apnea.


DIFFERENTIAL IMPACT OF PERIODONTAL TREATMENT STRATEGIES DURING PREGNANCY ON PERINATAL OUTCOMES: A SYSTEMATIC REVIEW AND META-ANALYSIS.
This study aimed to evaluate the impact of different periodontal treatment strategies during pregnancy on perinatal outcomes. This systematic review and meta-analysis of clinical trials was conducted according to PRISMA guidelines to assess the effect of mouthwash in addition to scaling and root planning (SRPM) on pregnancy outcomes, including preterm birth, low birth weight, gestational age, and birth weight. Pooled risk ratios (RR), mean differences (MD), and 95% confidence intervals (CI) were calculated using the random effect model. Twenty trials involving 5938 participants, including thirteen trials comparing scaling and root planning (SRP) and seven trials comparing SRPM with control groups. SRPM was associated with reduced risk of preterm birth (RR = 0.37; 95%CI = 0.16-0.84; P = .017; I2=93.26%; P < .001; number needed to treat (NNT): 3), low birth weight (RR = 0.54; 95%CI = 0.40-0.74; P < .0001; I2 = 0%; P = .46; NNT: 13), increased gestational age (MD = 0.78; 95%CI: 0.19-1.37; P = .009; I2 = 87.15%; P < .001), and birth weight (MD = 121.77; 95%CI = 3.19-240.34; P = .044; I2 = 80.68%; P < .001). There were no statistically significant differences in the analysis of SRP group, except for the increased birth weight (MD = 93.85; 95% CI = 3.27-184.42; P = .042; I2 = 84.11%; P < .001). Using mouthwash in addition to scaling and root planning (SRPM) for the treatment of periodontal disease during pregnancy significantly improves perinatal outcomes.


Convergent Adaptation in Mitochondria of Phylogenetically Distant Birds: Does it Exist?
In a wide range of taxa, proteins encoded by mitochondrial genomes are involved in adaptation to lifestyle that requires oxygen starvation or elevation of metabolism rate. It remains poorly understood to what extent adaptation to similar conditions is associated with parallel changes in these proteins. We search for a genetic signal of parallel or convergent evolution in recurrent molecular adaptation to high altitude, migration, diving, wintering, unusual flight abilities, or loss of flight in mitochondrial genomes of birds. Developing on previous work, we design an approach for the detection of recurrent coincident changes in genotype and phenotype, indicative of an association between the two. We describe a number of candidate sites involved in recurrent adaptation in ND genes. However, we find that the majority of convergence events can be explained by random coincidences without invoking adaptation.


Anticonvulsants in anxiety disorders.
Anticonvulsant drugs have shown promising results in the treatment of mood disorders, leading to the investigation of their potential efficacy in other psychiatric disorders. Numerous case reports, open-label trials, and placebo- controlled trials investigating the use of anticonvulsants in the treatment of anxiety disorders have yielded broad and varying results. Generally, these studies have indicated a potential role for anticonvulsants in the treatment of anxiety as monotherapy or augmentation therapy; however, these early findings must be supported by additional investigation in large-scale, placebo-controlled studies. This article reviews past and current research being done in this novel area of psychopharmacology.


Outcomes of transplantation with partial T-cell depletion of matched or mismatched unrelated or partially matched related donor bone marrow in children and adolescents with leukemias.
Graft-versus-host disease (GVHD) remains a major barrier to successful hematopoietic stem cell transplant for patients who lack a matched related donor. Partial T-cell depletion (TCD) of the graft may decrease the risk of severe GVHD with unrelated donors (URD) and partially matched related donors (PMRD) while retaining an antileukemic effect. We analyzed our experience using URD and PMRD for pediatric patients with leukemias from 1990 to 2001. A subgroup of 'matched' URD donor pairs was retrospectively analyzed for high-resolution class I. Partial TCD was accomplished with monoclonal antibody T10B9 or OKT3 and complement. There were 76 URD (45% matched) and 28 PMRD recipients. Event-free survival (EFS) was 38.3%, and overall survival (OS) 45.1% at 3 years. On multivariate analysis, there was no difference in survival based upon marrow source, but nonrelapse mortality was higher with the use of PMRD. Relapse occurred in 6% of ALL patients, and 22.8% of AML/MDS patients. Grades III-IV GVHD was observed in only 6.7% of patients. Partial TCD allows use of matched or mismatched URD, or PMRD with little mortality from GVHD, durable engraftment, and no increase in relapse risk.


High salt culture conditions suppress proliferation of rat C6 glioma cell by arresting cell-cycle progression at S-phase.
As one of the in vitro model experiments for investigating a possible effect of extracellular environmental stresses on glial cell proliferation, the influence of high salt culture conditions on the growth of rat C6 glioma cells was examined. Exposure to the culture medium containing high concentrations of NaCl reduced the number of viable cells in a concentration-dependent manner without any significant change in their viability. In contrast, proliferation of these cells was not substantially altered by culturing them in hyperosmotic medium containing either sucrose or glycerol, both of which were osmotically almost equivalent to high salt culture medium. On the other hand, expression of the egr-1 gene, an immediate early gene related to the proliferation and differentiation of various cells, was enhanced by culturing glioma cells in high salt medium while the reduction of glial fibrillary acidic protein content, an index of glial cell differentiation, was observed under the same culture conditions. Further studies on the relationship between egr-1 gene expression and the cell cycle showed that cell-cycle progression was arrested at S-phase by culturing glioma cells in high salt medium. Moreover, both resveratrol and CPT-11, which are known to arrest cell-cycle progression, elevated egr-1 mRNA levels in glioma cells. Thus, these observations suggest that high salt culture conditions might suppress the proliferation of rat C6 glioma cells as a consequence of arresting cell-cycle progression at S-phase, resulting secondarily in the compensatory enhancement of egr-1 gene expression.


Obesity impairs skeletal muscle repair through NID-1 mediated extracellular matrix remodeling by mesenchymal progenitors.
Obesity triggers skeletal muscle physio-pathological alterations. However, the crosstalk between adipose tissue and myogenic cells remains poorly understood during obesity. We identified NID-1 among the adipose tissue secreted factors impairing myogenic potential of human myoblasts and murine muscle stem cells in vitro. Mice under High Fat Diet (HFD) displayed increased NID-1 expression in the skeletal muscle endomysium associated with intramuscular fat adipose tissue expansion and compromised muscle stem cell function. We show that NID-1 is highly secreted by skeletal muscle fibro-adipogenic/mesenchymal progenitors (FAPs) during obesity. We demonstrate that increased muscle NID-1 impairs muscle stem cells proliferation and primes the fibrogenic differentiation of FAPs, giving rise to an excessive deposition of extracellular matrix. Finally, we propose a model in which obesity leads to skeletal muscle extracellular matrix remodeling by FAPs, mediating the alteration of myogenic function by adipose tissue and highlighting the key role of NID-1 in the crosstalk between adipose tissue and skeletal muscle.


Effects of calcium on the heterogeneity of the Na+, K+-ATPase forms in rat heart.
The sensitivity of the Na+, K+-ATPase to ouabain has been studied in sarcolemma vesicles isolated from normal rat heart. Two enzyme forms exhibiting high and low sensitivities to ouabain have been observed in Ca2+-free perfused heart. The half-maximal inhibitory effects occurred with 1-2 X 10(-8) M ouabain. The high sensitivity form undetectable in hearts maintained at a physiological Ca2+ level might represent altered low affinity sites or latent enzyme forms unmasked by low calcium concentrations. The heterogeneity of the Na+, K+-ATPase forms was found to be also modulated by the K+/ouabain antagonism, addition of K+ accentuating the heterogeneity. These in vitro results associated with in vivo experiments on isolated rat heart working under isovolumic conditions suggested that lowering Ca2+ has qualitative and quantitative effects. Low Ca2+ concentrations increased the sensitivities to ouabain and the amplitudes of both the enzyme inhibition and the positive inotropic effects.


Zinc alleviates mercury-induced oxidative stress in Pfaffia glomerata (Spreng.) Pedersen.
The possible role of zinc (Zn) to reverse the oxidative stress caused by mercury (Hg) was investigated in Pfaffia glomerata plantlets. Thirty-day-old acclimatized plantlets of P. glomerata were exposed to four treatments: control, 50 μM Zn, 50 μM Hg and 50 μM Zn + 50 μM Hg for 9 days. In Zn + Hg treatment, shoot and root Hg concentrations were 59 and 24% smaller than that plants exposed to 50 μM Hg added alone. An increase in the Zn concentration in the shoot of plants exposed to Zn + Hg occurred, although in the roots Zn concentration was not altered, when compared to the control. Fresh and dry weights, as well as the activity of δ-aminolevulinic acid dehydratase (δ-ALA-D) in Hg-treated plants were significantly reduced. Percentage survival, fresh and dry weights and δ-ALA-D activity of plants treated by 50 μM Zn + 50 μM Hg were greater than of that treated by Hg alone. Moreover, Zn treatment reduced the lipid peroxidation caused by Hg, being this effect related to increased root superoxide dismutase activity, and shoot catalase and ascorbate peroxidase activities. In conclusion, the presence of Zn in the substrate caused a significant reduction in the oxidative stress induced by Hg.


Behavioural flexibility of children with Autism Spectrum Disorder on a card-sorting task with varying task difficulty.
Inflexibility is taken to be a key characteristic of Autism Spectrum Disorder (ASD), although it is unclear which aspect of cognitive functioning is critical in this context. The current study investigated task-switching problems and inflexibility with a group of children with ASD, and a mental-aged matched control group. Participants (n = 50; mean age = 7 years) completed two card-sorting tasks, which involved learning to sort by either two or three possible dimensions, and then the sorting rule was switched although the number of dimensions required to sort the cards remained the same. Following the sorting rule change, the ASD group made more errors compared to controls. Errors were also related to task type (two or three dimensions), but this was not found to interact with ASD. If poor performance were solely dependent on executive function (working memory) problems in ASD, then a steeper decrease in performance with an increase in task difficulty for one group, compared to another group, would be expected. The current results suggest that task difficulty is an aspect of importance in set-shifting, but shifting is not differentially affected by this component.


First serological evidence of Crimean-Congo haemorrhagic fever virus in transhumant bovines in Italy.
Crimean-Congo haemorrhagic fever (CCHF) is an emerging tick-borne disease caused by the arbovirus Crimean-Congo haemorrhagic fever virus (CCHFV; family Nairoviridae). Given the public health impact, CCHF is considered a priority disease for the European Union. This study describes the first detection of anti-CCHFV antibodies in transhumant bovines in Italy. Sera from 794 cattle collected across Basilicata region (Southern Italy) were screened using a commercial ELISA kit. The animal-level and herd-level seroprevalences detected were 1.89% [95%CI: 1.12-3.1] and 29.63% [95%CI: 15.68-48.65], respectively. Results of the χ2 test for trend show that the exposure to CCHFV was significantly associated with increasing age, with the odds 5 times higher in 11-22-year old cattle than 1-4-year old cattle. The detection of antibodies against CCHFV in indigenous cattle indicates that the infection occurred in the study area and may warrant further consideration. Additionally, no significant spatial clustering of CCHF infection was detected, supporting the hypothesis that the disease is widespread in the region. Further studies at larger scale are needed to identify the areas at higher risk of zoonotic infection. A One Health approach should be implemented to better understand the disease risk and dynamics in the country, which effectively address the related public health threat.


[Importance of orientation on visual and neuronal resolution (author's transl)].
Spatial resolution of gratings of different orientation has been evaluated in 39 patients, by means of projected slides and by laser interference fringes. Both methods reveal a significantly higher resolution (about 12%) for vertical or horizontal bars compared to oblique ones. The similarity of findings for both methods is indicative of the neuronal origin of this orientation-dependent effect and exclude an optical origin.


A double-blind, placebo-controlled study of the safety and efficacy of ipratropium bromide nasal spray versus placebo in patients with the common cold.
Ipratropium bromide (IB) has been found to reduce secretions in the upper respiratory tract; this is accomplished through competitive inhibition of acetylcholine at muscarinic receptors that control rhinorrhea production. This study compared the safety and efficacy of IB with placebo in the symptomatic relief of rhinorrhea in patients with the common cold. Human subjects with symptoms of a common cold, primarily rhinorrhea, were enrolled and treated with either IB (84 micrograms/nostril) or placebo; each was administered as two sprays per nostril, four times a day, for 4 days. Primary efficacy analyses were in-clinic measurements of nasal discharge weights over a 3-hour period after administration on days 1 and 2 and assessment of rhinorrhea symptoms by use of a subjective patient-completed visual analog rating scale. IB significantly reduced rhinorrhea an average of 18% over placebo for days 1 and 2 (p = 0.01). Visual analog scale scores showed an average improvement in rhinorrhea of 22% over placebo (p = 0.001). When patients with relatively minor rhinorrhea (baseline weight of nasal discharge < or = 1.0 gm) were excluded, IB produced an average reduction in nasal discharge of 23% over placebo for days 1 and 2 (p = 0.003).


A new surgical approach of temporary ovarian transposition for children undergoing brachytherapy: technical assessment and dose evaluation.
We developed a new technique of temporary ovarian transposition (OT) for prepubertal girls undergoing brachytherapy. The aim of this study was to describe it, assess its feasibility and safety and calculate the dose delivered to the ovary in order to prove its efficacy. Sixteen prepubertal patients underwent temporary OT for brachytherapy at our center from March 2001 to December 2012. OT was done either by laparotomy or by laparoscopy. In all patients, the ovaries were grasped with an atraumatic forceps and mobilized above the iliac crest level as high as possible without any dissection or division of the ovarian ligaments or of the fallopian tube. They were sutured to the anterior abdominal wall by a transfixing stitch of non-dissolvable suture knotted on the outside of the patient on a pledget. Median age at surgery was 3 years (range: 2-9 years). The integrity of the fallopian tube was respected and not a single ligament was dissected or divided. None of the patients had intraoperative or postoperative complications. The stitches were retrieved after completion of irradiation and the ovaries in all the patients fell back into the pelvis. The calculated median radiation dose to the ovary was 1.4 Gy (range: 0.4-2.4 Gy). This surgical technique is simple and safe, either by laparotomy or by laparoscopy. It meets the radiation and physical constraints in prepubertal girls with vaginal or bladder RMS. However, longer follow-up is required to assess the ovarian function.


Talking with the chronic patient about malnutrition
We talk with Alejandro Toledo, the President of the Fundación Renal ALCER and Vicepresident of the First Platform of Patients Organizations.


Erratum to: Incidental Findings of Intense Radioiodine Uptake in Struma Ovarii and Bilateral Non-Lactating Breast Simultaneously on Post-Ablation SPECT/CT for Papillary Thyroid Cancer.
[This corrects the article DOI: 10.1007/s13139-016-0459-0.].


Natural product drug discovery: the successful optimization of ISP-1 and halichondrin B.
The concept that natural products provide excellent leads for drug discovery, ultimately producing viable drugs, is a widely accepted view. Natural products embody inherent structural complexity and biological activity which often leads to new targets, pathways, or modes of action. The challenge lies in identifying quality natural product scaffolds that can ultimately result in a drug. Two recently approved drugs originating from unlikely natural product leads, ISP-1 and halichondrin B, were examples of such high quality scaffolds. In initial testing, both compounds displayed excellent in vitro potency, but more importantly were amenable to chemical optimization. This combination of unique biological activity plus the generation of structural activity relationships (SAR) may be early indicators of a high quality natural product scaffold worthy of additional studies.


Bacterial dynamics in a raw cow's milk Caciotta cheese manufactured with aqueous extract of Cynara cardunculus dried flowers.
To investigate the bacterial dynamics of a Caciotta cheese traditionally manufactured in the Montefeltro area (Central Italy) with raw cow's milk and an aqueous extract of dried flowers from Cynara cardunculus as a coagulating agent. Conventional methods and a combined PCR-DGGE approach, relying on culture-dependent and -independent analyses, were used to investigate the cheese bacterial community, with a special focus on lactic acid bacteria. A heterogeneous population, including enterococci, lactococci, lactobacilli, food spoilage and other banal micro-organisms, was found. The study contributed to highlighting the influence of different technological parameters on bacterial dynamics of a raw milk Caciotta cheese coagulated with vegetable rennet. None of the species found in the vegetable rennet became dominant during the cheese-making and a prevailing role of the adventitions microbita coming from the raw milk and the dairy environment was highlighted.


Protocol for the trial to establish a causal linkage between mycotoxin exposure and child stunting: a cluster randomized trial.
The number of stunted children has fallen globally but continues to increase in Africa. Stunting is estimated to contribute to 14-17% of child deaths under 5 years of age and is a risk factor for poor cognitive and motor development and educational outcomes. Inadequate dietary intake and disease are thought to be the immediate causes of undernutrition and stunting. However, improving infant diets through complementary feeding interventions has been shown to only modestly reduce stunting. Multiple observational studies demonstrate a dose response relationship between fetal and post-natal aflatoxin exposure and reduced linear growth. This community-based cluster randomized trial will measure the effect of a reduced aflatoxin diet on length-for-age Z scores at 18 months in central Tanzania. All 52 health facilities in the Kongwa District of Dodoma Region were randomized into two groups. Starting at 6 months of age, participants in the intervention group receive a low-aflatoxin pre-blended porridge flour containing maize and groundnut (ratio 4:1 respectively) and low-aflatoxin groundnut flour, whereas in the control group the same porridge mix and groundnut flour are promoted through education but acquired by the household. Both groups will receive the same infant and young child feeding education and a thermos flask. A total of 3120 infants between 6 weeks and 3 months of age will be recruited into the study over 1 year. Data will be collected four times - at recruitment and when the infants are 6, 12 and 18 months of age. In a cohort of 600 infants, additional data will be collected at 9 and 15 months of age. The primary outcome is length-for-age at 18 months. Secondary outcomes include the Z scores for weight-for-age, middle upper arm circumference and head circumference, and the blood biomarker aflatoxin-albumin in the full sample, with the urine biomarker aflatoxin M1 analyzed in the cohort only. Better understanding the etiology of childhood stunting can lead to more appropriate interventions and policies to further reduce linear growth faltering and meet the Sustainable Development Goals. NCT03940547, (April 24, 2019).


Modifications of the FIM instrument under the inpatient rehabilitation facility prospective payment system.
To describe the modifications made to the FIM instrument when it was incorporated into the Inpatient Rehabilitation Facility-Patient Assessment Instrument (IRF-PAI), and to compare FIM data collected before and after the IRF prospective payment system (IRF-PPS) was implemented in 2002 for patients with stroke. Year-by-year comparison of data of Medicare patients with stroke discharged in 1998-2003 from 411 IRFs that submitted data to the Uniform Data System for Medical Rehabilitation for each of those years. In the pre-PPS period, admission motor FIM ratings decreased slightly, and discharge motor, admission cognitive, and discharge cognitive ratings remained stable. Between 2001 and 2003, all four ratings decreased: admission motor by 1.8 FIM units, discharge motor by 3.3 FIM units, and admission and discharge cognitive each by 1.0 FIM unit. The lower admission FIM ratings led to an increase in the mean case-mix index from 1.39 to 1.49. The decrease in FIM ratings in the IRF-PAI/PPS years may reflect alterations in coding practices as a result of changed rules for rating the FIM instrument, "downcoding" leading to assignment into higher-paying categories, changes in the IRF patient population, and/or changes in IRF patient outcomes. Coding changes should be considered when comparing pre-PPS and PPS FIM data.


Analysis of the Temporal Patterning of Notch Downstream Targets during Drosophila melanogaster Egg Chamber Development.
Living organisms require complex signaling interactions and proper regulation of these interactions to influence biological processes. Of these complex networks, one of the most distinguished is the Notch pathway. Dysregulation of this pathway often results in defects during organismal development and can be a causative mechanism for initiation and progression of cancer. Despite previous research entailing the importance of this signaling pathway and the organismal processes that it is involved in, less is known concerning the major Notch downstream targets, especially the onset and sequence in which they are modulated during normal development. As timing of regulation may be linked to many biological processes, we investigated and established a model of temporal patterning of major Notch downstream targets including broad, cut, and hindsight during Drosophila melanogaster egg chamber development. We confirmed the sequential order of Broad upregulation, Hindsight upregulation, and Cut downregulation. In addition, we showed that Notch signaling could be activated at stage 4, one stage earlier than the stage 5, a previously long-held belief. However, our further mitotic marker analysis re-stated that mitotic cycle continues until stage 5. Through our study, we once again validated the effectiveness and reliability of our MATLAB toolbox designed to systematically identify egg chamber stages based on area size, ratio, and additional morphological characteristics.


Effects of macronutrient intake on sleep duration and quality: A systematic review.
The aim of this systematic review was to characterise and synthesise the literature that explores the effects of macronutrient manipulation on sleep outcomes in order to provide dietary recommendations for sleep improvement. Intervention studies that evaluated the effects of macronutrient manipulations on sleep were identified using four databases. A study was included if it: (a) delivered a macronutrient-based dietary intervention, regardless of length; (b) included healthy human participants aged 18 and older; and (c) measured some aspect of sleep, for example, duration or quality, in some manner. A total of 17 papers and 19 studies met the inclusion criteria. Included studies were organised into acute studies lasting less than 24 hours (n = 4), longer-term (≥24 hours) studies (n = 10), and energy restriction studies (n = 5). The greatest amount of support was observed for longer-term administration of higher carbohydrate diets. These interventions positively influenced rapid eye movement sleep; however, non-rapid eye movement sleep was negatively affected. The overall health outcomes of these changes are difficult to assess. Limited evidence suggests higher protein diets under conditions of energy restriction can improve sleep quality, but these effects might be restricted to individuals with overweight or obesity. Current evidence does not support the effects of acute macronutrient manipulation, defined as less than 24 hours, on sleep outcomes. Manipulating carbohydrate intake for at least 24 hours appears to alter sleep outcomes among healthy individuals. Interventions were highly varied and methodological shortcomings were identified. More work is required to fully understand how macronutrient intake affects sleep outcomes.


A review on biosurfactant producing bacteria for remediation of petroleum contaminated soils.
The discharge of potentially toxic petroleum hydrocarbons into the environment has been a matter of concern, as these organic pollutants accumulate in many ecosystems due to their hydrophobicity and low bioavailability. Petroleum hydrocarbons are neurotoxic and carcinogenic organic pollutants, extremely harmful to human and environmental health. Traditional treatment methods for removing hydrocarbons from polluted areas, including various mechanical and chemical strategies, are ineffective and costly. However, many indigenous microorganisms in soil and water can utilise hydrocarbon compounds as sources of carbon and energy and hence, can be employed to degrade hydrocarbon contaminants. Therefore, bioremediation using bacteria that degrade petroleum hydrocarbons is commonly viewed as an environmentally acceptable and effective method. The efficacy of bioremediation can be boosted further by using potential biosurfactant-producing microorganisms, as biosurfactants reduce surface tension, promote emulsification and micelle formation, making hydrocarbons bio-available for microbial breakdown. Further, introducing nanoparticles can improve the solubility of hydrophobic hydrocarbons as well as microbial synthesis of biosurfactants, hence establishing a favourable environment for microbial breakdown of these chemicals. The review provides insights into the role of microbes in the bioremediation of soils contaminated with petroleum hydrocarbons and emphasises the significance of biosurfactants and potential biosurfactant-producing bacteria. The review partly focusses on how nanotechnology is being employed in different critical bioremediation processes.


Prenatal impairment of brain serotonergic transmission in infants.
To evaluate whether the free fraction of L-tryptophan (L-Trp) and the N1/P2 component of the auditory evoked potentials (AEPs) are associated with impaired brain serotonin neurotransmission in infants with intrauterine growth restriction (IUGR). We measured free, bound, and total plasma L-Trp and recorded the N1/P2 component of AEP in a prospective, longitudinal, and comparative study comparing IUGR and control infants. Plasma free L-Trp was increased and the amplitude of N1/P2 component was significantly decreased in IUGR relative to control infants. The free fraction of L-Trp and N1/P2 component had a negative association. In newborns with IUGR, the changes in measured plasma free fraction of L-Trp and in the amplitude the N1/P2 component of the AEP suggest an inverse association between free L-Trp and components of the AEP. The changes observed in the free fraction of L-Trp and AEP may be causally associated with brain serotonergic activity in utero. In IUGR, epigenetic factors such as stress-induced disturbances in brain serotonin metabolism or serotonergic activity, identifiable by alterations in AEP, influence cerebral sensory cortex development and may be causally associated with serotonin-related disorders in adulthood.


Stenting for malignant colonic obstruction: a comparison of efficacy and complications in colonic versus extracolonic malignancy.
Self-expandable metal stent placement for palliation of malignant colonic obstruction for colorectal cancer (CRC) is safe and efficacious. In contrast, outcomes of stent placement for extracolonic malignancy (ECM) are unclear. To compare the success and complication rates of colorectal stenting in patients with CRC versus those with ECM. Retrospective chart review. Tertiary-care academic medical center. Between September 2000 and December 2007, all patients with malignant colon obstruction in whom endoscopy was performed with the intention of placing a colonic metal stent. Technical and clinical success rates, surgical interventions, and procedure-related complications. Colonic stenting was performed for CRC in 34 patients and for ECM in 15 patients. Patients with CRC were more likely to have clinical success after all endoscopic therapy (94.1%) than those with ECM (20.0%) (P < .0001). Surgical diversion to relieve persistent obstructive symptoms was required in significantly more patients with ECM. Five patients with ECM (33.3%) had at least one complication, including 2 deaths, compared with 3 patients with CRC (8.8%) (P = .046). Only underlying ECM was predictive of failed colon stent placement by multivariate analysis (hazard ratio 21.0, P = .0013). A history of radiation therapy was the sole predictor of complications (hazard ratio 7.8, P = .048). Single institution, retrospective analysis, relatively small sample size. Colon stenting for large-bowel obstruction from ECM is infrequently successful and is associated with a significantly higher risk of complications in comparison with patients with CRC.


Hepatic Induction of Fatty Acid Binding Protein 4 Plays a Pathogenic Role in Sepsis in Mice.
Sepsis is defined as the host's deleterious systemic inflammatory response to microbial infections. Herein, we report an essential role of the fatty acid binding protein 4 (FABP4; alias adipocyte protein 2 or aP2), a lipid-binding chaperone, in sepsis response. Bioinformatic analysis of the Gene Expression Omnibus data sets showed the level of FABP4 was higher in the nonsurvival sepsis patients' whole blood compared to the survival cohorts. The expression of Fabp4 was induced in a liver-specific manner in cecal ligation and puncture (CLP) and lipopolysaccharide treatment models of sepsis. The induction of Fabp4 may have played a pathogenic role, because ectopic expression of Fabp4 in the liver sensitized mice to CLP-induced inflammatory response and worsened the animal's survival. In contrast, pharmacological inhibition of Fabp4 markedly alleviated the CLP responsive inflammation and tissue damage and improved survival. We conclude that FABP4 is an important mediator of the sepsis response. Early intervention by pharmacological inhibition of FABP4 may help to manage sepsis in the clinic.


The effects of breaks on digital eye strain, dry eye and binocular vision: Testing the 20-20-20 rule.
To evaluate the benefits on the eyes of taking breaks based on the 20-20-20 rule. Bespoke computer software using the laptop webcam to assess user breaks, eye gaze and blinking, and emitting personalized reminders of breaks based on the 20-20-20 rule, was downloaded onto the laptops of 29 symptomatic computer users. Digital eye strain (DES), binocular vision and dry eye were assessed before and after two weeks of using the reminders and one week after the discontinuation of the strategy. Binocular measurements included visual acuity, accommodative posture, stereopsis, fixation disparity, ocular alignment, accommodative facility, positive/negative vergences and near point of convergence. Symptoms were evaluated using the computer vision syndrome questionnaire, ocular surface disease index (OSDI), and symptom assessment in dry eye questionnaire (SANDE) versions one and two. Dry eye signs were assessed by measuring tear meniscus height, conjunctival redness, blink rate and incomplete blinking, lipid layer thickness, non-invasive keratograph break-up time, corneal and conjunctival staining and lid wiper epitheliopathy. A decrease in the duration of computer work and the duration of breaks, along with an increase in the number of breaks taken per day was observed as a result of the 20-20-20 rule reminders (p ≤ 0.015). No changes on any binocular parameter were observed after the management period (p ≥ 0.051), except for an increase in accommodative facility (p = 0.010). Dry eye symptoms and DES decreased with the rule reminders (p ≤ 0.045), although this improvement was not maintained one week after discontinuation (p > 0.05). No changes on any ocular surface and tear film parameter were observed with the rule reminders (p ≥ 0.089). The 20-20-20 rule is an effective strategy for reducing DES and dry eye symptoms, although 2 weeks was not enough to considerably improve binocular vision or dry eye signs.


Mental health, functional impairment, and barriers to mental health access among cancer patients in Vietnam.
This study evaluated the prevalence and severity of depression and anxiety symptomatology, barriers to mental health access, and correlates of functional impairment among cancer inpatients. This cross-sectional study recruited adult cancer patients (N = 300) in June and July 2022 at the largest oncological hospital in Vietnam. Multivariable linear regression analyses examined the association between demographics, clinical characteristics, and patients' functional impairment. Approximately 46.3% and 27.0% showed some depression and anxiety symptomatology, while 8.0% and 3.0% experienced major depressive and anxiety symptoms, respectively. Patients reported the most impairment in mobility and capacity for life activities. More functional impairment was identified in patients with gastrointestinal cancers, those receiving radiation therapy alone, and those scoring higher on depression and anxiety than in those with cancers originating in the head, neck, or lung or those receiving chemotherapy alone. Reports of better overall health status were negatively associated with functional impairment. Patients reported extensive perceived barriers to seeking psychiatric care, including not knowing where to get mental health support (86.7%), wanting to manage mental health independently (73.7%), and thinking mental health will resolve on its own (73.7%), and denying mental health concerns (61.0%). High frequency and severity of depression and anxiety symptomatology underscore the importance of integrating mental health services into existing oncological treatment protocols. Increasing mental health literacy and provision of psychoeducation is critical to addressing barriers to mental health service access. Integration of functional impairment evaluations into hospital admission and discharge planning is also needed.


End-of-Life Care among Nursing Home Residents with Dementia Varies by Nursing Home and Market Characteristics.
Nursing homes (NHs) are critical end-of-life (EOL) care settings for 70% of Americans dying with Alzheimer's disease/related dementias (ADRD). Whether EOL care/outcomes vary by NH/market characteristics for this population is unknown but essential information for improving NH EOL care/outcomes. Our objectives were to examine variations in EOL care/outcomes among decedents with ADRD and identify associations with NH/market characteristics. Cross-sectional. Place-of-death (hospital/NH), presence of pressure ulcers, potentially avoidable hospitalizations (PAHs), and hospice use at EOL. Key covariates were ownership, staffing, presence of Alzheimer's units, and market competition. Long-stay NH residents with ADRD, age 65 + years of age, who died in 2017 (N = 191,435; 14,618 NHs) in NHs or hospitals shortly after NH discharge. National Medicare claims, Minimum Data Set, public datasets. Descriptive analyses and multivariable logistic regressions. As ADRD severity increased, adjusted rates of in-hospital deaths and PAHs decreased (17.0% to 6.3%; 11.2% to 7.0%); adjusted rates of dying with pressure ulcers and hospice use increased (8.2% to 13.5%; 24.5% to 40.7%). Decedents with moderate and severe ADRD had 16% and 13% higher likelihoods of in-hospital deaths in for-profit NHs. In NHs with Alzheimer's units, likelihoods of in-hospital deaths, dying with pressure ulcers, and PAHs were significantly lower. As ADRD severity increased, higher licensed nurse staffing was associated with 14%‒27% lower likelihoods of PAHs. Increased NH market competition was associated with higher likelihood of hospice use, and lower likelihood of in-hospital deaths among decedents with moderate ADRD. Decedents with ADRD in NHs that were nonprofit, had Alzheimer's units, higher licensed nurse staffing, and in more competitive markets, had better EOL care/outcomes. Modifications to state Medicaid NH payments may promote better EOL care/outcomes for this population. Future research to understand NH care practices associated with presence of Alzheimer's units is warranted to identify mechanisms possibly promoting higher-quality EOL care.


Anorectal manometry findings in relation with long-term functional outcomes of the patients operated on for Hirschsprung's disease compared to the reference-based population.
This study investigated anorectal manometry (AM) findings and bowel function of patients operated on for Hirschsprung's disease (HD). A cross-sectional study was conducted at Children's Hospital 2. Patients operated on for HD from January 2015 to January 2020 were reviewed. Their clinical characteristics, bowel function, and manometric findings were investigated and compared with the references. Ninety-five patients and 95 references were enrolled. Mean ages were 6.6 ± 2.2 years and 7.2 ± 2.9 years,; fecal incontinence rates were 25.3% and 2.1%, and constipation rates were 12.6% and 4.2 for the patients versus the references, respectively. Anal resting pressures were significantly decreased in the patients compared to the references (53.2 ± 16.1 mmHg versus 62.2 ± 14.0 mmHg; p < 0.05). Among the patients, the anal resting pressure was significantly decreased in the incontinents than in the continents (46.0 ± 10.6 mmHg versus 55.6 ± 16.9 mmHg, p < 0.05). During the sensation test, the value of maximum tolerated volume was significantly decreased in the incontinents than in the continents (135.9 ± 47.9 mL versus 166.6 ± 58.3 mL, p < 0.05). AM is an objective method providing beneficial information that could guide a more adapted management in HD patients with defecation disorders.


Long-term trend in serum (1,3)-β-D-glucan level in a man with chronic disseminated candidiasis treated with corticosteroids.
Chronic disseminated candidiasis (CDC) is a type of invasive candidiasis. CDC commonly appears in the neutrophil recovery phase after chemotherapy in patients with hematologic malignancies, and immune reconstitution inflammatory syndrome (IRIS) is thought to play a major role in CDC development. This report describes the case of a 33-year-old man with CDC as a complication of acute myeloid leukemia. We describe the clinical course, body temperature, therapy, and (1,3)-β-D-glucan (BDG) levels over the course of 22 months. He was initially treated with antifungals, but corticosteroids were added because of a persistently elevated body temperature, which we attributed to IRIS. After starting corticosteroids, his clinical condition improved, but his BDG levels became markedly elevated. We hypothesize that the suppression of the excessive immune response by corticosteroids lead to granuloma collapse, fungal release, and hematogenous dissemination, resulting in elevated BDG levels. The patient's condition gradually improved over the course of follow-up.


A modified mixed haemadsorption procedure for the detection and localisation of sperm antigens.
A mixed haemadsorption procedure for the detection and localisation of spermatozoal antigens is described. Sperm are immobilised by attaching them to the bases of wells on plastic plates using poly-L-lysine and antisera and sensitised red cells are then added to these wells. The technique uses simple reagents and allows sperm/red cell rosettes to be kept as permanent records.


One-by-one immediate dental implants: A papillae preservation concept for adjacent implants in a compromised periodontal case.
In an immediate implant placement and provisionalization strategy, the esthetic results of multiple adjacent implants can be obtained even with compromised periodontium by implementing the "one-by-one" protocol. Staged extractions of multiple adjacent teeth to maintain soft tissue architecture are a key feature of the technique described.


Hierarchy of cytotoxic T cell clones. II. Intraclonal triggering of lysis by hapten-specific CTL.
Cells from clones of anti-hapten murine cytotoxic T lymphocytes (CTL) can act as both target and effector cells, but will not lyse members of the same clone. The effect of haptenation on the cytolytic activity of anti-fluorescein (FL) and anti-trinitrophenol (TNP) CTL clones was examined. Treatment of anti-FL clones with fluorescein isothiocyanate or anti-TNP clones with trinitrobenzene sulphonic acid induces these clones to kill in an antigen-independent fashion. Targets killed by the haptenated CTL included syngeneic and allogeneic B lymphocyte blast cells, P815, YAC-1 and in one case human GM 4072 tumor cells. The importance of CD8 and T cell receptor (TCR) occupancy is demonstrated by the ability to block autotriggering by antibody directed against Ly 2 and the TCR. The results demonstrate that effects other than antigen recognition of the target play a role in the final outcome of effector-target cell interactions and provide a mechanism which could lead to autodestruction and immunosuppression particularly in some types of viral infection.


The autologous stump plasty. Treatment for bony overgrowth in juvenile amputees.
Autologous stump capping is a procedure designed to prevent bony overgrowth in skeletally immature amputation stumps. All 19 capping procedures in the lower extremities were successful after an average follow-up of 7.3 years. All patients use their prostheses, and no secondary operations have been needed for stump problems. Of the 31 cap-plasties of the humerus, six required re-operation. The overall failure rate of 12% is low compared with the failure rate of re-amputation.


Carbapenem resistance in a human clinical isolate identified to be closely related to Acinetobacter indicus.
Here we report a case of carbapenem resistance in a human clinical isolate that was found to be closely related to the newly described environmental species Acinetobacter indicus. This strain harboured the blaOXA-23 carbapenemase gene located on a conjugative plasmid. Partial sequencing of 16S rDNA and rpoB genes, together with matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) analysis, showed that this strain was distantly related to the Acinetobacter baumannii-calcoaceticus complex and was closely related to A. indicus.


How native yeasts may influence the chemical profile of the Brazilian spirit, cachaça?
Cachaça is the typical Brazilian spirit, obtained by distillation of fermented sugarcane must, whose production is closely related to regional heritage and geographical traits. In this work, we investigate the influence of different autochthonous yeast species, in comparison to other commercial strains, on the chemical profile of cachaça. Fermentations were performed with four different wild species, a mixed starter culture with native yeasts, and two commercial strains. Procedures were carried out simulating traditional practices, the distillates analyzed by gas chromatography, and data analyzed by multivariate statistics. Results show that the overgrowth of some non-Saccharomyces during fermentation can lead to higher concentration of contaminant compounds in the distillate. However, those microorganisms do not impress negative traits when working in cluster association, and may indeed contribute to the distinctive chemical quality and flavor of cachaça. Spontaneous fermentation are still the traditional method used for the production of cachaça, and our results indeed suggest that higher diversity, and evened fermentative environments, enable yeasts to act as a complex cluster and imprint a distinctive, quality chemical profile to the distillate. Finally, this study may help to understand the role of the native microbiota and its influence on cachaça's chemical and sensory profiles, besides demonstrating the importance of adopting insightful handling practices and strict control of technical guidelines.


Hydrogen-deuterium exchange mass spectrometry highlights conformational changes induced by factor XI activation and binding of factor IX to factor XIa.
Factor XI (FXI) is a zymogen in the coagulation pathway that, once activated, promotes haemostasis by activating factor IX (FIX). Substitution studies using apple domains of the homologous protein prekallikrein have identified that FIX binds to the apple 3 domain of FXI. However, the molecular changes upon activation of FXI or binding of FIX to FXIa have remained largely unresolved. This study aimed to gain more insight in the FXI activation mechanism by identifying the molecular differences between FXI and FXIa, and in the conformational changes in FXIa induced by binding of FIX. Hydrogen-deuterium exchange mass spectrometry was performed on FXI, FXIa, and FXIa in complex with FIX. Both activation and binding to FIX induced conformational changes at the interface between the catalytic domain and the apple domains of FXI(a)-more specifically at the loops connecting the apple domains. Moreover, introduction of FIX uniquely induced a reduction of deuterium uptake in the beginning of the apple 3 domain. We propose that the conformational changes of the catalytic domain upon activation increase the accessibility to the apple 3 domain to enable FIX binding. Moreover, our HDX MS results support the location of the proposed FIX binding site at the beginning of the apple 3 domain and suggest a mediating role in FIX binding for both loops adjacent to the apple 3 domain.


LOCATIONAL AGREEMENT OF NEAR-INFRARED AUTOFLUORESCENCE WITH CHOROIDAL VASCULAR HYPERPERMEABILITY IN CENTRAL SEROUS CHORIORETINOPATHY.
To characterize the findings of eyes with choroidal vascular hyperpermeability (CVH), a hallmark of central serous chorioretinopathy, using multimodal imaging and investigate the locational agreement between these findings and CVH. Among patients with central serous chorioretinopathy in either eye, eyes with CVH identified using indocyanine green angiography without exudative changes were included. All eyes were examined using funduscopy, fluorescein angiography, spectral-domain optical coherence tomography, and short-wavelength or near-infrared autofluorescence (SWAF or NIRAF). The locational agreement between CVH and imaging findings was evaluated for each modality. The relative index on how they overlapped was calculated as the overlapping index. Binarized images, particularly for NIRAF, were also evaluated. This study included 69 CVH sites in 33 eyes of 28 patients. Pachydrusen was detected in 36% of CVH sites. Fluorescein angiography revealed hyperfluorescent areas in 39% of CVH sites. Optical coherence tomography findings identified 65% of CVH sites, but the overlapping index was 5%. Short-wavelength or near-infrared autofluorescence imaging identified 89% of CVH sites, but they exhibited variable autofluorescence. NIRAF imaging revealed hypoautofluorescence findings in all CVH sites. When binarized, near-infrared autofluorescent dark dots were observed in all CVH sites. Overlapping indices before and after binarization were 78% and 98%, respectively. The near-infrared autofluorescent dark dots area corresponded well with that of CVH (intraclass correlation coefficient, 0.987 [95% confidence interval, 0.952-0.995]). Hypoautofluorescent findings revealed on NIRAF imaging, especially after binarization processing, corresponded well with CVH sites. These multimodal imaging results may help investigate the anatomical or locational characteristics of CVH in patients with central serous chorioretinopathy.


Electronic structure, cationic, and excited states of nitrogen-containing spiroborates.
This paper presents the results of the study of the electronic structure and cationic and excited states of three spiroborate complexes (2-acetylacetonato-1,3,2-benzodioxaborol, its NH- and NMe-derivatives) and three corresponding ligands (acetylacetone, 4-aminopent-3-en-2-one, and 4-methylaminopent-3-en-2-one). Materials based on spiroborates are used in medicine, for example, as a drug carrier. In industry, spiroborate anions are used in ionic liquids and as alternative high performance lubricants. Analysis of experimental and calculated data allowed determining the influence of functional groups on the parameters of the electronic structure and energy of electronic transitions. Compared to acetylacetone and its NH- and NMe-derivatives, the upper filled molecular orbitals of the corresponding spiroborates are stabilized at 0.4-1.7 eV, which is due to the positive charge of the ligand due to the acceptor properties of the dioxyphenylene fragment. Among the studied compounds, when replacing the oxygen atom in the α-position with the NH- or NMe-group, a bathochromic shift of intense bands in the absorption spectra is observed, since the energy intervals between the orbitals of the π3 and π4 ligand are reduced. In addition, in a number of spiroborates, the violation of C2v symmetry when replacing an oxygen atom leads to the appearance of a low-intensity maximum in the long-wave part of the absorption spectrum, due to the π2X → π4 transition. Complexes were studied by photoelectron spectroscopy, absorption spectroscopy, and high-level ab initio quantum chemical computations, including the algebraic diagrammatic construction method for the polarization propagator of the second order (ADC(2)), the outer-valence Green's function (OVGF), the density functional theory (DFT), the time-dependent density functional theory (TDDFT) and the domain-based local pair natural orbital (EOM-DLPNO) methods. X-ray photoelectronic spectra of two spiroborates in the condensed state were measured using a two-chamber high-vacuum system MXPS XP (Omicron, Germany). UV-visible absorption spectra were recorded using a spectrophotometer 2550 (Shimadzu-UV, Japan). The geometry of all studied compounds was optimized by the DFT/B3LYP/Def2-SVP method. The energy of electron levels in the S0 state and the distribution of electron density at each MO were obtained by the DFT/CAMB3LYP/cc-pVDZ method. The energies of excited states were obtained by the TDDFT/CAMB3LYP/cc-pVDZ, ADC(2)/cc-pVDZ and EOM-DLPNO/cc-pVDZ methods. All DFT and TDDFT calculations were carried out in the GAMESS (US) software computing package. ADC(2) calculations of excited states were performed using the Orca 4.0.1 software package. EOM-DLPNO and OVGF calculations were carried out in the Gaussian 16 software package.


Real-time monitoring of the release of multiple payloads from nanomaterials.
We overcome limitations of conventional methods to monitor the release of two payloads in situ. The concentration of two different corrosion inhibitors are simultaneously determined during their release from nanofibers by square wave voltammetry (SWV). SWV is suitable for direct and simultaneous determination of concentration of two payloads.


Can FDA-Approved Immunomodulatory Drugs be Repurposed/Repositioned to Alleviate Chronic Pain?
Pain is among the most widespread chronic health condition confronting society today and our inability to manage chronic pain contributes to the opioid abuse epidemic in America. The immune system is known to contribute to acute and chronic pain, but only limited therapeutic treatments such as non-steroid anti-inflammatory drugs have resulted from this knowledge. The last decade has shed light on neuro-immune interactions mediating the development, maintenance, and resolution of chronic pain. Here, we do not aim to perform a comprehensive review of all immune mechanisms involved in chronic pain, but to briefly review the contribution of the main cytokines and immune cells (macrophages, microglia, mast cells and T cells) to chronic pain. Given the urgent need to address the Pain crisis, we provocatively propose to repurpose/reposition FDA-approved immunomodulatory drugs for their potential to alleviate chronic pain. Repositioning or repurposing offers an attractive way to accelerate the arrival of new analgesics.


The profiling and identification of the absorbed constituents and metabolites of Naoshuantong capsule in mice biofluids and brain by ultra- fast liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry.
Naoshuantong capsule (NSTC) is an oral traditional Chinese medicine formula used widely in the clinic for ischemic stroke. The absorbed ingredients and metabolites of NSTC have never been reported before. In this study, a method incorporating rapid resolution liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to identify absorbed ingredients and metabolites after oral administration of NSTC. A total of 15 constituents were detected and identified as prototypes of NSTC. 109 metabolites related to catechin, gallic acid, paeoniflorin, chlorogenic acid, protocatechuate, typhaneoside, β-elemene, calycosin were identified in serum, urine and brain. 19 metabolites of typhaneoside, 3 metabolites of β-elemene, 12 metabolites of calycosin were reported for the first time. This is the first time to explore the absorption and metabolism of NSTC. The work will provide helpful information for further research of the mechanism and application of NSTC.


Density-functional study of the structures and properties of holmium-doped silicon clusters HoSi n (n = 3-9) and their anions.
The structures and properties of Ho-doped Si clusters, including their adiabatic electron affinities (AEAs), simulated photoelectron spectra (PESs), stabilities, magnetic moments, and charge-transfer characteristics, were systematically investigated using four density-functional methods. The results show that the double-hybrid functional (which includes an MP2 correlation component) can accurately predict the ground-state structure and properties of Ho-doped Si clusters. The ground-state structures of HoSi n (n = 3-9) are sextuplet electronic states. The structures of these Ho-doped Si clusters (aside from HoSi7) are substitutional. The ground-state structures of HoSi n- are quintuplet electronic states. Their predicted AEAs are in excellent agreement with the experimental ones. The mean absolute error in the theoretical AEAs of HoSi n (n = 4-9) is only 0.04 eV. The simulated PESs for HoSi n- (n = 5-9) are in good agreement with the experimental PESs. Based on its simulated PES and theoretical AEA, we reassigned the experimental PES of HoSi4- and obtained an experimental AEA of 2.2 ± 0.1 eV. The dissociation energies of Ho from HoSi n and HoSi n- (n = 3-9) were evaluated to test the relative stabilities of the clusters. HOMO-LUMO gap analysis indicated that doping the Si clusters with the rare-earth metal atom significantly increases their photochemical reactivity. Natural population analysis showed that the magnetic moments of HoSi n (n = 3-9) and their anions derive mainly from the Ho atom. It was also found that the magnetic moments of Ho in the HoSi n clusters are larger than the magnetic moment of an isolated Ho atom.


Detubulation experiments localise delayed rectifier currents to the surface membrane of amphibian skeletal muscle fibres.
Ionic currents in intact and detubulated frog sartorius muscle fibres were compared at room temperature using a loose-patch voltage clamp configuration in four experimental groups. The test fibres (i) were detubulated by a previously established osmotic shock protocol that involved the introduction and withdrawal of extracellular glycerol followed by exposure to Ca2+/Mg2+-Ringer solution and cooling. The control fibres were spared osmotic shock and (ii) simply studied in normal Ringer solution, (iii) exposed to 30 min of steady cooling to 9-10 degrees C before electrophysiological study or (iv) exposed to and studied in glycerol-Ringer solution. The presence or absence of detubulation was confirmed for all the experimental groups through assessing for the abolition or otherwise of the delayed after-depolarisation normally associated with action potential propagation into the transverse (T) tubules. All fibre groups showed similar resting potentials (-80 to -90 mV) thus ensuring consistent baseline voltages from which the voltage clamp steps were imposed. The intact muscle fibres in the three control groups (ii)-(iv) spared osmotic shock showed both inward Na+ and delayed rectifier outward (K+) currents. In contrast, patches from detubulated muscle fibres in the test group (i) showed only delayed outward currents, consistent with contrasting contributions to Na+ and K+ currents from regions of membrane affected or spared by the detubulation procedure. Nevertheless, the voltage dependence, maximum steady state amplitudes and timecourses of the delayed outward currents were conserved through all the experimental groups. These findings suggest that the surface as opposed to the tubular membrane contributes the greater part of the delayed rectifier current in amphibian skeletal muscle.


Flavouring composition prepared by fermentation with Bacillus spp.
Fermented foodstuffs and condiments remain a key constituents of diets throughout many parts of Asia and Africa. In cases where the process of fermentation evolved for the development of taste or aroma, it often resulted in enhanced nutrition, stabilisation of the original raw materials, and detoxification of anti-nutrient factors. Several fermented products rely on the participation of various Bacillus species, including Bacillus natto and B. subtilis. Often, the finished products are of a very local character and exhibit sensory properties resulting from unique flora and processing technologies applied in small scale, home-based fermentations. Fermentation with B. natto and B. subtilis can produce very characteristic aromas in fermented products such as natto and dawadawa (also referred to as daddawa). Moreover, the hydrolytic capabilities of these microorganisms can result in a precursor-rich environment, which is useful for subsequent reactions leading to flavour production. A 1995 patented process demonstrated the ability to produce a fermented flavouring composition with the use of Bacillus spp. Hydrolysed protein obtained after fermentation with Bacillus spp. is mixed with reactive flavour precursors, which are subsequently heated to induce flavour formation and can be dried to a powder format. The product of this patented process imparts a basic meaty flavour, with a reduced yet characteristic dawadawa-like aroma. This paper briefly summarises some of the characteristics and uses of traditional dawadawa and illustrates alternatives described in the patent for the production of a process flavour base. Issues and considerations for the industrialisation of a fermentation process are briefly discussed, as well as some future opportunities for development and exploitation of traditional fermentation technology.


Rapid spectral scan and stopped-flow studies of carbon monoxide binding to bovine adrenocortical cytochrome P-450scc.
Carbon monoxide binding with both cholesterol-free (low-spin) and cholesterol-bound (high-spin) reduced forms of purified cytochrome P-450scc has been investigated by rapid-scan and stopped-flow spectrometry. CO binding occurs within 150 ms at 25 degrees C for both forms of P-450scc, with a typical absorption maximum at 450 nm. Isosbestic points occur at the following wavelengths: between reduced-CO and reduced cholesterol-free P-450scc at 434 and 471 nm; between reduced-CO and reduced cholesterol-bound P-450scc at 433 and 469 nm. Both the 'on' (k1) and 'off' rate constants (k-1) are found to be independent of pH between pH 5 and 9. The mean values of k1 for cholesterol-free (1.8 +/- 0.2) X 10(5) M-1 X s-1) and cholesterol-bound [1.9 +/- 0.1) X 10(5) M-1 X s-1) P-450scc are almost identical, while the mean value of k-1 for the former [2.3 +/- 0.3) X 10 s-1) is about double that of the latter [1.2 +/- 0.1) X 10 s-1). This suggests the instability of the reduced-CO complex in the absence of cholesterol.


The toxicology of Amanita virosa: the destroying angel.
This paper examines the biology and medical consequences of ingesting the potentially lethal poisonous mushroom, Amanita virosa, the Destroying Angel. The fungus, its structure, distribution and toxic components are described. Symptoms of human poisoning by A. virosa are described, following the order of Homeopathic Repertories. Laboratory values for comparison with normal values of haematology, biochemistry and urine analyses are given.


Physical activity and mortality in men and women with diagnosed cardiovascular disease.
Existing information regarding the optimal types and amount of physical activity for secondary prevention is controversial. We examined the association between different types of physical activity (domestic, walking, sports) and mortality in participants with established cardiovascular disease (CVD). Data were collected from the Scottish Health Surveys (1995, 1998, 2003) that were linked to a patient-based database of CVD hospital admissions and deaths. There were 837 men and women with clinically confirmed CVD at baseline, of whom 175 died over an average of 5.6+/-3.1 years of follow-up. The lowest risks for all-cause mortality were seen in participants undertaking at least 20 min/week of sports activity (fully adjusted hazard ratio=0.32, 95% confidence interval=0.16-0.65) or walking (hazard ratio=0.74, 0.53-1.04). Similar associations were observed for CVD mortality. The physically active also showed lower levels of inflammatory risk markers and greater HDL cholesterol. Men and women with existing CVD who participate in moderate-to-vigorous activity for at least 20 min/week showed the lowest risks of all-cause and cardiovascular mortality. These findings support emerging data from controlled exercise training studies.


The absence of crossovers on chromosome 4 in Drosophila melanogaster: Imperfection or interesting exception?
Drosophila melanogaster chromosome 4 is an anomaly because of its small size, chromatin structure, and most notably its lack of crossing over during meiosis. Earlier ideas about the absence of crossovers on 4 hypothesize that these unique characteristics function to prevent crossovers. Here, we explore hypotheses about the absence of crossovers on 4, how these have been addressed, and new insights into the mechanism behind this suppression. We review recently published results that indicate that global crossover patterning, in particular the centromere effect, make a major contribution to the prevention of crossovers on 4.


Integral valorisation of walnut shells based on a three-step sequential delignification.
Walnut kernels represent no more than 50-60% of the total weight of the fruit, so the sum of walnut shells generated every year is immense. Nonetheless, these shells could be further valorised for the extraction of their main constituents following a biorefinery scheme. Hence, the objective of this work was an integral valorisation of walnut shells, which involved a sequential organosolv delignification (200 °C, 90 min, 70/30 v/v EtOH/H2O, LSR 6:1) and several posterior non-isothermal hydrothermal treatments (180, 195 and 210 °C, LSR 8:1). Moreover, the spent solids after the aforementioned treatments were evaluated as possible sources of cellulose nanocrystals. The results showed that the sequential organosolv delignifications presented relative lignin yields up to 60%, which leaded to lignins that just differed on their molecular weight distributions. The hydrothermal treatments were efficient for the removal of still present hemicelluloses (14.7-71.8%), and permitted a successful cellulose nanocrystal obtaining whereas the spent solid from the delignification stages did not. Thus, this study presented an innovative strategy for the integral valorisation of walnut shells.


Lung colonization and metastasis by disseminated B16 melanoma cells: H-2 associated control at the level of the host and the tumor cell.
We have studied the experimental metastasis of H-2+ and H-2- melanoma sublines in H-2b/b and H-2a/b hosts by enumerating pulmonary colonies 20-50 days after i.v. inoculation of tumor cells. In H-2b/b hosts, the H-2+ "B16-S" cells gave rise to a moderate number of metastatic colonies (mean: 6.3 +/- 6). The "BL16-L" sublines that had lost the expression of MHC class-I antigens, according to FACS-analysis and quantitative absorption tests, gave no metastases under the same conditions. Pretreatment of the H-2+ met+ B16-S with interferons (beta or alpha + beta) increased their H-2 antigen expression and the number of metastatic colonies (mean: 25 +/- 16). Interferon pretreatment of B16-L cells partially restored their H-2b expression and induced them to form a small number of metastatic colonies. The reduction in pulmonary colonization by the H-2 negative B16-L cells could be attributed to their rapid elimination by natural killer cells, already observed within 24 hr of inoculation of radiolabelled cells. H-2- B16-L cells were more susceptible than H-2+ B16-S cells to in vitro lysis by poly I:C-treated splenocytes, and they acquired full metastatic abilities if the hosts were treated with anti-asialo GM-I serum. In H-2a/b heterozygous hosts, the H-2+ B16-S cells also failed to metastasize. Reduced pulmonary colonization was evident by 24 hr after injection in comparison with H-2b/b hosts, and could be reversed by anti-asialo GM-I treatment of the hosts. In vitro, H-2a/b splenocytes were more cytotoxic to the B16 cells than syngeneic effectors. The results are discussed in relation to a recent hypothesis on a surveillance mechanism for elimination of cells on the basis of their lack (or insufficient expression) of host MHC genes.


Refining personality assessments by combining MCMI high point profiles and MCMI codes. Part III. MMPI code 24/42.
In this study it has been shown that the addition of MCMI high point code data to the MMPI 24/42 code type serves to clarify contradictory MMPI descriptors and produces three distinct clusters. These MMPI/MCMI clusters comprise an interpersonally acting-out group, an interpersonally acting-in group, and an emotionally acting-out group. These groups were useful in determining different roles for the MMPI 2 and 4 scales insofar as they relate to both personality (Axis II) and its clinical consequences (Axis I). The results of this study also lend support to the utility of combining objective assessment instruments both for identifying and for discriminating among relevant personality disorders and clinical syndromes.


What is the Impact of Racial Disparities on Diagnosis and Receipt of Appropriate Mental Health Care Among Urology Patients?
Patients with chronic disease and mental illness are at higher risk of depression and suicide. Many who have been diagnosed with genitourinary cancers are at higher risk of suicide, even among those who have sought out mental health services. Under-represented populations (African-American, Hispanic, elderly, disabled) suffer disproportionately from a lack of mental health services. However, not much is reported on the interplay of mental health and genitourinary cancer in these populations. This review aims to identify the relevant literature and describe a path forward to address and alleviate this disparity. PATIENT SUMMARY: Patients with chronic disease and mental illness are at higher risk of depression and suicide. Little is known about the complex interplay between race and mental health in patients with urologic cancers and more research is needed.


Nitrile Synthesis with Aldoxime Dehydratases: A Biocatalytic Platform with Applications in Asymmetric Synthesis, Bulk Chemicals, and Biorefineries.
Nitriles comprise a broad group of chemicals that are currently being industrially produced and used in fine chemicals and pharmaceuticals, as well as in bulk applications, polymer chemistry, solvents, etc. Aldoxime dehydratases catalyze the cyanide-free synthesis of nitriles starting from aldoximes under mild conditions, holding potential to become sustainable alternatives for industrial processes. Different aldoxime dehydratases accept a broad range of aldoximes with impressive high substrate loadings of up to >1 Kg L-1 and can efficiently catalyze the reaction in aqueous media as well as in non-aqueous systems, such as organic solvents and solvent-free (neat substrates). This paper provides an overview of the recent developments in this field with emphasis on strategies that may be of relevance for industry and sustainability. When possible, potential links to biorefineries and to the use of biogenic raw materials are discussed.


Role of B₁₂ and homocysteine status in determining BMD and bone turnover in young Indians.
Vitamin B(12) (B(12)) deficiency and hyperhomocysteinemia (HHcy) are independent risk factors for low bone mineral density (BMD) and fracture risk. We studied the role of HHcy and B(12) deficiency in determining the peak bone mass in Indians. Randomly selected 151 healthy young adult subjects (females 100, mean age: 26 yr) underwent evaluation of dietary intake of calcium and B(12); sun exposure; estimation of BMD by dual-energy X-ray absorptiometry at total hip, forearm, and lumbar spine; serum 25(OH)D(3); intact parathyroid hormone; B(12); homocysteine (Hcy); and bone turnover markers (BTMs) serum crosslaps, N-mid osteocalcin, and bone-specific alkaline phosphatase. Hypovitaminosis D (serum 25OHD(3)<20 ng/mL) and serum ALP level >150 IU/L were seen in 83% and 27%, respectively. Median serum B(12) and Hcy levels were 140 pg/mL (interquartile range [IQR]: 72-230 pg/mL) and 18 μmol/L (IQR 14-32 μmol/L); B(12) deficiency (serum B(12)<200 pg/mL) and HHcy (serum Hcy>30 μmol/L) were present in 71% and 68%, respectively. Low BMD (Z-score <-2.0) was present in 17% of subjects. There was no significant correlation between serum Hcy, folate, B(12), BTM, and BMD. BMD was predicted by height, weight, and body mass index. Young Indian healthy adults have high prevalence of hypovitaminosis D, B(12) deficiency, and HHcy. There is no correlation of serum B(12), folate, and Hcy status with BTMs and BMD in young, healthy, vegetarian Indian adults. Anthropometric variables predict BMD in young Indians.


Aberrant promoter methylation of the CHD1 gene may contribute to the pathogenesis of breast cancer: a meta-analysis.
Cadherin-1 (CHD1), as an invasion suppressor gene, could suppress tumor cell invasion and metastasis in various tumors, but reduced CHD1 levels, resulting from epigenetic silencing, are common in poorly differentiated, advanced stage carcinomas. This meta-analysis was performed to evaluate the relationships between promoter methylation of CHD1 and breast cancer. Relevant studies were retrieved from the Web of Science (1945 ~ 2013), the Cochrane Library (Issue 12, 2013), PubMed (1966 ~ 2013), EMBASE (1980 ~ 2013), CINAHL (1982 ~ 2013), and the Chinese Biomedical Database (CBM) (1982 ~ 2013) using a systematic literature search. Results were summarized by meta-analyses, conducted using the STATA software (version 12.0, Stata Corporation, College Station, TX, USA). Odds ratios (ORs) and 95 % confidence intervals (95 % CIs) were calculated. In the present meta-analysis, 9 cohort studies with a total of 425 patients with breast cancer were included. Our meta-analysis results demonstrated that the frequency of CHD1 promoter methylation in cancer tissues was significantly higher than that in normal tissues, adjacent tissues, and benign tissues (cancer tissue vs. normal tissue OR = 30.87, 95 % CI = 16.76 ~ 56.86, P < 0.001; cancer tissue vs. adjacent tissue OR = 23.30, 95 % CI = 12.85 ~ 42.26, P < 0.001; cancer tissue vs. benign tissue OR = 2.94, 95 % CI = 1.60 ~ 5.40, P < 0.001; respectively). Ethnicity-stratified analysis indicated that aberrant CHD1 promoter methylation was strongly correlated with breast cancer among both Asians and Caucasians in the majority of subgroups. Our results suggest that aberrant promoter methylation of the CHD1 gene may have a high frequency in breast cancer tissues. Thus, CHD1 methylation could be correlated with the pathogenesis of breast cancer.


Using bimetallic Au/Cu nanoplatelets for construction of facile and label-free inner filter effect-based photoluminescence sensing platform for sarcosine detection.
Herein, we report a facile and label-free method for sensitive and specific determination of prostate cancer biomarker sarcosine via using photoluminescent bimetallic Au/Cu nanoplatelets (AuCu NPs) to construct an inner filter effect (IFE)-based photoluminescence (PL) sensing platform. The AuCu NPs were formed by the cysteine-induced co-reduction reaction, which displayed bright PL with an emission peak at 560 nm. Meanwhile, the Cu(I) doping caused a maximum 25-fold enhancement of quantum yield (QY), compared with the native Au(I) complexes, i.e., from 0.85 to 21.5%. By integrating the AuCu NPs with p-phenylenediamine (PPD) oxidation reaction, an IFE-based sensor for sarcosine detection was constructed. In this method, sarcosine is oxidized under the catalysis of sarcosine oxidase (SOx) to yield H2O2. The latter further oxidizes PPD to form 2,5-diamino-N,N'-bis(p-aminophenyl)-l,4-benzoquinone di-imine (PPDox) in the presence of horseradish peroxidase (HRP). The UV-vis absorption spectrum of the PPDox can overlap well with the excitation and emission spectra of the AuCu NPs, resulting in the efficient quenching of the AuCu NPs via the IFE effect. Therefore, this IFE-based AuCu NPs/SOx/PPD/HRP sensing platform can be used for highly sensitive and specific sensing of sarcosine. The sensing platform showed two linear regions of the PL intensity of the AuCu NPs versus the concentration of sarcosine in the range of 0.5-5 μM and 5-100 μM with a detection limit (LOD) of 0.12 μM (S/N = 3). Furthermore, this IFE-based sensing platform could be developed into a paper-based biosensor for simple, instrument-free, and visual detection of sarcosine.


Extraction of diclofenac by SiO2 -NH2 @Fe3 O4 and its determination: Central composite design.
In this work, magnetic nanoparticles (Fe3 O4 ) were prepared by simple co-precipitation method in aqueous medium and then subsequently modified with tetraethyl orthosilicate and 3-aminopropyl triethylenesilane. The properties of the particles were characterized by FTIR spectroscopy X-ray diffraction, transmission electron microscopy, and scanning electron microscopy. The SiO2 -NH2 @Fe3 O4 particles were successfully applied to simultaneously enrich and separate diclofenac from water, urine, and plasma samples. The method, which takes the advantages of both nanoparticle adsorption and magnetic phase separation from the sample solution, could avoid some of the time-consuming experimental procedures related to the traditional solid phase extraction. The main parameters affecting extraction and desorption efficiency such as pH, amount of SiO2 -NH2 @Fe3 O4 , volume of desorption solvent, and extraction time were screened. The significant variables were optimized by using central composite design. At optimum conditions values of variables set as pH = 4, 10 mg SiO2 -NH2 @Fe3 O4 , 0.5 mL methanol, and 15 min extraction time and then the extracted diclofenac were injected to HPLC for analysis. The linear response (r2 > 0.9992) was obtained in the range of 0.004-15 µg/mL with detection limit 0.0012 µg/mL and extraction recovery was in the range of 92-96% with RSD < 5% (n = 6).


Efficacy of afoxolaner (NexGard®) on the treatment of myiasis caused by the New World screwworm fly Cochliomyia hominivorax (Diptera: Calliphoridae) in naturally infested dogs.
New World screwworm (NWS) myiasis is an infestation by Cochliomyia hominivorax larvae that consume the living tissue of warm-blooded animals, including humans. Domestic dogs are among the potential hosts of these flies that lay their eggs on the edges of wounds. NWS myiasis cases can be fatal if untreated. Treatment with parasiticides must be fast-acting, long-lasting and show 100% efficacy, since open wounds can be reinfested. Afoxolaner is a molecule from the isoxazoline family with proven ectoparasiticide action against fleas, ticks and mites in dogs. Fourteen healthy client-owned dogs, naturally infested by C. hominivorax larvae, were treated with afoxolaner (NexGard®) as per label recommendations, providing at least the minimum dosage of 2.5 mg/kg. Maggot infestations were classified as light (fewer than 10 larvae), mild (from 10 to 20 larvae) or severe (more than 20 larvae), according to the number of larvae found in the wound and/or collected from the ground after treatment. Twenty-four hours post-treatment, infested lesions were carefully inspected and collected larvae were counted and classified as live or dead. All maggots were identified as second and third instar larvae of C. hominivorax and were found dead within 24 h after treatment, demonstrating 100% larvicidal efficacy against C. hominivorax.


Accuracy of reduction and early clinical outcome in acetabular fractures treated by the standard ilio-inguinal versus the Stoppa/iliac approaches.
In acetabular fractures, the correct choice of the surgical approach is mandatory to achieve accurate reduction and to avoid complications. Anterior approaches include the ilio-inguinal, the Stoppa, the ilio-femoral and the para-rectal exposures. The first two are the most commonly used approaches nowadays. The aim of this study was to compare these two approaches. The standard three window ilio-inguinal approach was compared to the intra-pelvic Stoppa approach with an added iliac window. The study enrolled 54 patients. Patients were divided into two groups. The first group consisted of 33 patients presented with acetabular fractures and had ORIF starting with an ilio-inguinal exposure. This group was compared to a second group of 21 patients who were treated with the Stoppa/iliac window approach. All patients were treated by one surgical team. The accuracy of reduction, the early clinical results and the approach related complications were compared.


Long persistence of rigor mortis at constant low temperature.
We studied the persistence of rigor mortis by using physical manipulation. We tested the mobility of the knee on 146 corpses kept under refrigeration at Torino's city mortuary at a constant temperature of +4 degrees C. We found a persistence of complete rigor lasting for 10 days in all the cadavers we kept under observation; and in one case, rigor lasted for 16 days. Between the 11th and the 17th days, a progressively increasing number of corpses showed a change from complete into partial rigor (characterized by partial bending of the articulation). After the 17th day, all the remaining corpses showed partial rigor and in the two cadavers that were kept under observation "à outrance" we found the absolute resolution of rigor mortis occurred on the 28th day. Our results prove that it is possible to find a persistence of rigor mortis that is much longer than the expected when environmental conditions resemble average outdoor winter temperatures in temperate zones. Therefore, this datum must be considered when a corpse is found in those environmental conditions so that when estimating the time of death, we are not misled by the long persistence of rigor mortis.