Atg5- and Atg7-dependent autophagy in dopaminergic neurons regulates cellular and behavioral responses to morphine.
The molecular basis of chronic morphine exposure remains unknown. In this study, we hypothesized that macroautophagy/autophagy of dopaminergic neurons would mediate the alterations of neuronal dendritic morphology and behavioral responses induced by morphine. Chronic morphine exposure caused Atg5 (autophagy-related 5)- and Atg7 (autophagy-related 7)-dependent and dopaminergic neuron-specific autophagy resulting in decreased neuron dendritic spines and the onset of addictive behaviors. In cultured primary midbrain neurons, morphine treatment significantly reduced total dendritic length and complexity, and this effect could be reversed by knockdown of Atg5 or Atg7. Mice deficient for Atg5 or Atg7 specifically in the dopaminergic neurons were less sensitive to developing a morphine reward response, behavioral sensitization, analgesic tolerance and physical dependence compared to wild-type mice. Taken together, our findings suggested that the Atg5- and Atg7-dependent autophagy of dopaminergic neurons contributed to cellular and behavioral responses to morphine and may have implications for the future treatment of drug addiction.


Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping.
Sequences of antigenic determinants were identified by limited proteolysis of peptide antigens bound to an immobilized monoclonal antibody and direct molecular weight determination of the monoclonal antibody-bound peptide fragments by 252Cf plasma desorption mass spectrometry. The epitope peptides to the monoclonal antibody h453 [Burger, R., Zilow, G., Bader, A., Friedlein, A. & Naser, W. (1988) J. Immunol. 141, 553-558] were isolated from immobilized antigen-antibody complexes by partial trypsin digestion. A synthetic eicosapeptide comprised of the C-terminal sequence of the human complement component polypeptide des-Arg77-C3a as well as guinea pig des-Arg78-C3a was used as an antigen. Conditions were developed under which trypsin specifically degraded the antigens without inactivation of the immobilized antibody. After proteolysis, epitope peptides were dissociated from the antibody with 4 M MgCl2. The antigenic peptides were purified by HPLC and identified by 252Cf plasma desorption mass spectrometry. The epitope recognized by h453 resides on the C-terminal tryptic peptides of human (residues 70-76) and guinea pig (residues 70-77) C3a. As an estimation of accuracy this method is able to provide, trypsin digestion of immune complexes caused cleavage of the antigen within a distance of two amino acid residues upstream from the epitope.


[Effect of miR-125b on T Cell Activation in Aplastic Anemia by Targetting B7-H4].
To investigate the effect of miR-125b on T cell activation in patients with aplastic anemia (AA) and its molecular mechanism. A total of 30 AA patients were enrolled in department of hematology, Binzhou Medical University Hospital from January 2018 to October 2021, as well as 15 healthy individuals as healthy control (HC) group. Peripheral blood mononuclear cells (PBMCs) were isolated, in which the levels of miR-125b and B7-H4 mRNA were detected by RT-qPCR. Immunomagnetic beads were used to separate naive T cells and non-naive T cells from AA patients and healthy people to detect the levels of miR-125b and B7-H4 mRNA. Lentivirus LV-NC inhibitor and LV-miR-125b inhibitor were transfected into cells, and T cell activation was detected by flow cytometry. The dual-luciferase reporter gene assay was used to detect the targetting relationship between miR-125b and B7-H4. RT-qPCR and Western blot were used to detect the levels of miR-125b, CD40L, ICOS, IL-10 mRNA and B7-H4 protein. Compared with HC group, the expression of miR-125b was up-regulated but B7-H4 mRNA was down-regulated in PBMCs of AA patients (P <0.05), and the proportions of CD4+CD69+ T cells and CD8+CD69+ T cells in PBMCs of AA patients were higher (P <0.05). The expression of miR-125b was significantly up-regulated but B7-H4 mRNA was down-regulated in both naive T cells and non-naive T cells of AA patients (P <0.05), and non-naive T cells was more significant than naive T cells (P <0.05). Compared with NC inhibitor group, the expression of miR-125b was significantly decreased, the expression level of CD69 on CD4+ and CD8+ T cells in PBMCs was also significantly decreased, while the luciferase activity was significantly increased after co-transfection of miR-125b inhibitor and B7-H4-3'UTR-WT in the miR-125b inhibitor group (P <0.05). Compared with NC inhibitor group, the mRNA and protein levels of B7-H4 were significantly increased in the miR-125b inhibitor group (P <0.05). Compared with miR-125b inhibitor+shRNA group, the expression levels of CD69 on CD4+ and CD8+ T cells were significantly increased, and the levels of CD40L, ICOS and IL-10 mRNA were also significantly increased in the miR-125b inhibitor+sh-B7-H4 group (P <0.05). MiR-125b may promote T cell activation by targetting B7-H4 in AA patients.


COVID-19 and spinal cord injuries: The viewpoint from an emergency department resident with quadriplegia.
Spinal cord injuries (SCIs) present distinct physiological and social considerations for the emergency physician. During the COVID-19 pandemic, these considerations may generate unique challenges for emergency physicians managing patients with SCIs. Physiological disruptions may alter the way SCI patients present with COVID-19. The same disruptions can affect management of this vulnerable patient group, perhaps warranting early aggressive treatment. The medical picture will often be complicated by unique social characteristics. The reliance on caregivers for activities of daily living can, as an example, increase the human resource requirement of an ED. Considering the vulnerabilities and complexities of patients with SCI, the community should prioritise prevention of COVID-19 infections in this group. In the event that they do present to an ED, planning for and understanding their complexities will facilitate optimal management.


circ‑Grm1 promotes pulmonary artery smooth muscle cell proliferation and migration via suppression of GRM1 expression by FUS.
Pulmonary arterial hypertension is a progressive and fatal disease. Recent studies suggest that circular RNA (circRNAs/circs) can regulate various biological processes, including cell proliferation. Therefore, it is possible that circRNA may have important roles in pulmonary artery smooth muscle cell proliferation in hypoxic pulmonary hypertension (HPH). The aim of the present study was to determine the role and mechanism of circRNA‑glutamate metabotropic receptor 1 (circ‑Grm1; mmu_circ_0001907) in pulmonary artery smooth muscle cell (PASMC) proliferation and migration in HPH. High‑throughput transcriptome sequencing was used to screen circRNAs and targeted genes involved in HPH. Cell Counting Kit‑8 (CCK‑8), 5‑ethynyl‑2‑deoxyuridine and wound healing assays were employed to assess cell viability and migration. Reverse transcription‑quantitative PCR and western blotting were used to detect target gene expression in different groups. Bioinformatical approaches were used to predict the interaction probabilities of circ‑Grm1 and Grm1 with FUS RNA binding protein (FUS). The interactions of circ‑Grm1, Grm1 and FUS were evaluated using RNA silencing and RNA immunoprecipitation assays. The results demonstrated that circ‑Grm1 was upregulated in hypoxic PASMCs. Further experiments revealed that the knockdown of circ‑Grm1 could suppress the proliferation and migration of hypoxic PASMCs. Transcriptome sequencing revealed that Grm1 could be the target gene of circ‑Grm1. It was found that circ‑Grm1 could competitively bind to FUS and consequently downregulate Grm1. Moreover, Grm1 could inhibit the function of circ‑Grm1 by promoting the proliferative and migratory abilities of hypoxic PASMCs. The results also demonstrated that circ‑Grm1 influenced the biological functions of PASMCs via the Rap1/ERK pathway by regulating Grm1. Overall, the current results suggested that circ‑Grm1 was associated with HPH and promoted the proliferation and migration of PASMCs via suppression of Grm1 expression through FUS.


Improving the modeling of disease data from the government surveillance system: a case study on malaria in the Brazilian Amazon.
The study of the effect of large-scale drivers (e.g., climate) of human diseases typically relies on aggregate disease data collected by the government surveillance network. The usual approach to analyze these data, however, often ignores a) changes in the total number of individuals examined, b) the bias towards symptomatic individuals in routine government surveillance, and; c) the influence that observations can have on disease dynamics. Here, we highlight the consequences of ignoring the problems listed above and develop a novel modeling framework to circumvent them, which is illustrated using simulations and real malaria data. Our simulations reveal that trends in the number of disease cases do not necessarily imply similar trends in infection prevalence or incidence, due to the strong influence of concurrent changes in sampling effort. We also show that ignoring decreases in the pool of infected individuals due to the treatment of part of these individuals can hamper reliable inference on infection incidence. We propose a model that avoids these problems, being a compromise between phenomenological statistical models and mechanistic disease dynamics models; in particular, a cross-validation exercise reveals that it has better out-of-sample predictive performance than both of these alternative models. Our case study in the Brazilian Amazon reveals that infection prevalence was high in 2004-2008 (prevalence of 4% with 95% CI of 3-5%), with outbreaks (prevalence up to 18%) occurring during the dry season of the year. After this period, infection prevalence decreased substantially (0.9% with 95% CI of 0.8-1.1%), which is due to a large reduction in infection incidence (i.e., incidence in 2008-2010 was approximately one fifth of the incidence in 2004-2008).We believe that our approach to modeling government surveillance disease data will be useful to advance current understanding of large-scale drivers of several diseases.


[Functional and metabolic activity of peripheral blood neutrophils in acute viral hepatitis B and C].
The functional metabolic activity of peripheral blood neutrophils in acute virus hepatitis B (VHB) and/or virus hepatitis C (VHC) was evaluated. 48 patients were examined; of these, VHB was diagnosed in 28 patients and VHC was diagnosed in 9 patients and the mixed form of virus hepatitis (VHB + VHC), in 11 patients. Determination of adhesive capacity of neutrophils, production of superoxidase anion in the nitro blue tetrazolium (NBT) test, activity of myeloperoxidase (MPO) and acidic phosphatase (AP), the amount of cation proteins (CP) was made. Most pronounced functional dysbalance of neutrophil leukocytes and considerable changes in biochemical characteristics of the activity of the infectious process in patients with the mixed form of virus hepatitis were established. These data demonstrated that in acute virus hepatitis B and C at the peak of the disease such characteristics of the functional activity of neutrophils as results of the NBT test, the activity of MPO and AP, as well as the amount of CP, were highly informative.


Knowledge and attitudes about otitis media risk: implications for prevention.
To investigate maternal knowledge and attitudes about otitis media (OM) risk, to estimate the prevalence of risk factors in the first year of life, and to identify barriers to the reduction of risk factors (eg, formula feeding, day care attendance, and exposure to passive smoke). Questionnaires mailed to a systematic sample of 504 Minnesota women >/=18 years old identified through 1994 birth certificates. Eighty percent returned a completed survey. According to maternal report, 29% of infants (age 8 to 13 months) had recurrent OM (>/=3 episodes) and 2% had tympanostomy tubes. Forty-six percent attended day care, 29% had >/=1 smoking parent, and 49% breastfed for </=2 months. Women were more knowledgeable about OM signs and symptoms than about risk factors. Mean OM knowledge score (the sum of correct true-false responses) was 7.0 (standard deviation = 1.6). Using multiple linear regression, knowledge score was significantly related to marital status, education, age, area of residence, breastfeeding (months), and number of cigarettes smoked per day by the mother, but not to infant or sibling OM history or day care attendance (R = .23). Infant history of OM (odds ratio, 1.9; 95% confidence interval, 1.1 to 3.2) and white race (odds ratio, 0.3; 95% confidence interval, 0. 1 to 0.8), but not the presence of risk factors, were significantly related to having received clinicians' advice about OM prevention advice. OM education and prevention programs should target pregnant women and new mothers with OM risk factors, and those who are young, single, and less educated.


A biomimetic multi-layered collagen-based scaffold for osteochondral repair.
Cartilage and osteochondral defects pose a significant challenge in orthopedics. Tissue engineering has shown promise as a potential method for the treatment of such defects; however, a long-lasting repair strategy has yet to be realized. This study focuses on the development of a layered construct for osteochondral repair, fabricated through a novel "iterative layering" freeze-drying technique. The process involved repeated steps of layer addition followed by freeze-drying, enabling control over material composition, pore size and substrate stiffness in each region of the construct, while also achieving a seamlessly integrated layer structure. The novel construct developed mimics the inherent gradient structure of healthy osteochondral tissue: a bone layer composed of type I collagen and hydroxyapatite (HA), an intermediate layer composed of type I collagen, type II collagen and HA and a cartilaginous region composed of type I collagen, type II collagen and hyaluronic acid. The material properties were designed to provide the biological cues required to encourage infiltration of host cells from the bone marrow while the biomechanical properties were designed to provide an environment optimized to promote differentiation of these cells towards the required lineage in each region. This novel osteochondral graft was shown to have a seamlessly integrated layer structure, high levels of porosity (>97%), a homogeneous pore structure and a high degree of pore interconnectivity. Moreover, homogeneous cellular distribution throughout the entire construct was evident following in vitro culture, demonstrating the potential of this multi-layered scaffold as an advanced strategy for osteochondral defect repair.


Sufficient dimension reduction for populations with structured heterogeneity.
A key challenge in building effective regression models for large and diverse populations is accounting for patient heterogeneity. An example of such heterogeneity is in health system risk modeling efforts where different combinations of comorbidities fundamentally alter the relationship between covariates and health outcomes. Accounting for heterogeneity arising combinations of factors can yield more accurate and interpretable regression models. Yet, in the presence of high-dimensional covariates, accounting for this type of heterogeneity can exacerbate estimation difficulties even with large sample sizes. To handle these issues, we propose a flexible and interpretable risk modeling approach based on semiparametric sufficient dimension reduction. The approach accounts for patient heterogeneity, borrows strength in estimation across related subpopulations to improve both estimation efficiency and interpretability, and can serve as a useful exploratory tool or as a powerful predictive model. In simulated examples, we show that our approach often improves estimation performance in the presence of heterogeneity and is quite robust to deviations from its key underlying assumptions. We demonstrate our approach in an analysis of hospital admission risk for a large health system and demonstrate its predictive power when tested on further follow-up data.


A boy with normal growth in spite of growth hormone deficiency after resection of a suprasellar teratoma.
We reported a boy with panhypopituitarism after removal of a suprasellar teratoma and pituitary stalk transection at the age of 3 months. His growth was accelerated after 5 years of age without growth hormone (GH) therapy, although he had poor height growth until age 4 under treatment with hydrocortisone, levothyroxine sodium, and desamino-D-arginine vasopressin (DDAVP). Hyperphagia and obesity developed after surgery. Endocrinological examination revealed no GH response to glucagon, low serum levels of insulin-like growth factor (IGF)-1 and IGF binding protein-3 (IGFBP-3). Serum prolactin was normal, but serum insulin was high. Some patients who received an operation for craniopharyngioma were reported to achieve normal growth without GH secretion, but the mechanism is still unknown. High serum levels of prolactin or insulin can be associated with normal IGF in GH deficient patients. This patient had obesity and high serum insulin, which may be related to growth without GH secretion.


[Intelligence and neurocognitive tests among students living in a industrialized region of Sardinia with relatively low blood levels of lead].
In several recent epidemiological studies blood lead levels (BLLs) even below the current CDC intervention level of 10 microg/dl have been associated with reduced neurocognitive capacities of children, with no clear evidence of a "safe" threshold. We analyzed the relationship between the BLLs and the neurocognitive capacities of 205 Sardinian students aged 11 to 15 years, using 2 tests of the Swedish Performance Evaluating System (SPES) and the full-scale Intelligence Quotient (IQ) derived from the Wechsler Intelligence Scale for Children (WISC). The studied population included 104 children (61 males and 43 females) living in Portoscuso, a town 2 Km far from a lead smelter (mean BLLs: 5.98 +/- 2.2; max 11.5 microg/dl), and 101 age-matched students (55 males and 46 females) living in Sant'Antioco, a town about 20 Km far from the same smelter (mean BLLs: 2.08 +/- 0.8; max 4.5 microg/dl). Subjects with BLLs above 4 microg/dl performed worse in the SPES tests and scored about 5.0 points less on the full-scale IQ compared to the students with lower BLLs. The adjusted regression coefficients derived from the multivariate analysis showed that higher BLLs were significantly associated with worse performances in the SPES tests and with reduced IQ (0.94 points for each microg/dl of BLL). This study confirms the potential neurotoxicity of low-levels of lead suggesting the need of lowering the actual CDC "limit of concern" for children to values lower than 4 microg/dl, improving at the same time the environmental primary prevention for limiting the lead exposure of subjects living near the lead smelter.


Diosmin pretreatment improves cardiac function and suppresses oxidative stress in rat heart after ischemia/reperfusion.
Reperfusion of ischemic tissue leads to the generation of oxygen derived free radicals which plays an important role in cellular damage. Objective of the current study is to evaluate the cardio-protective and antioxidant effect of diosmin on ischemia-reperfusion related cardiac dysfunction, oxidative stress and apoptosis. Diosmin (50 and 100 mg/kg body weight (bw)) was given every day to the rats orally throughout the experimental period. Ischemia/reperfusion protocol was carried out ex vivo using langendorff perfusion method and the cardiac functional recovery was assessed in terms of percentage rate pressure product. Coronary effluents of LDH and CK-MB activities, antioxidant enzyme activities, lipid peroxidation products, activity of TCA cycle enzymes were evaluated. Moreover, in vitro superoxide anion and hydroxyl radical scavenging potential of diosmin was also quantified. Finally, quantitative real-time PCR was used for assessing Bcl-2 mRNA expression in heart. Cardiac functional recovery was impaired after reperfusion compared with continuously perfused heart. It was significantly prevented by diosmin treatment. Impaired antioxidant enzyme activities and elevated lipid peroxidation products level were also significantly suppressed. The activity of TCA cycle enzymes was protected against reperfusion stress. Down regulated Bcl-2 was also significantly increased. This study concluded that diosmin pretreatment prevents all the impaired patterns including cardiac function, oxidative stress and apoptosis associated with reperfusion in control heart by its antioxidant role.


Evolution of enzymatic activities in the enolase superfamily: N-succinylamino acid racemase and a new pathway for the irreversible conversion of D- to L-amino acids.
Members of the mechanistically diverse enolase superfamily catalyze reactions that are initiated by abstraction of the alpha-proton of a carboxylate anion to generate an enolate anion intermediate that is stabilized by coordination to a Mg2+ ion. The catalytic groups, ligands for an essential Mg2+ and acid/base catalysts, are located in the (beta/alpha)8-barrel domain of the bidomain proteins. The assigned physiological functions in the muconate lactonizing enzyme (MLE) subgroup (Lys acid/base catalysts at the ends of the second and sixth beta-strands in the barrel domain) are cycloisomerization (MLE), dehydration (o-succinylbenzoate synthase; OSBS), and epimerization (L-Ala-D/L-Glu epimerase). We previously studied a putatively promiscuous member of the MLE subgroup with uncertain physiological function from Amycolatopsis that was discovered based on its ability to catalyze the racemization of N-acylamino acids (N-acylamino acid racemase; NAAAR) but also catalyzes the OSBS reaction [OSBS/NAAAR; Palmer, D. R., Garrett, J. B., Sharma, V., Meganathan, R., Babbitt, P. C., and Gerlt, J. A. (1999) Biochemistry 38, 4252-4258]. In this manuscript, we report functional characterization of a homologue of this protein encoded by the genome of Geobacillus kaustophilus as well as two other proteins that are encoded by the same operon, a divergent member of the Gcn5-related N-acetyltransferase (GNAT) superfamily of enzymes whose members catalyze the transfer an acyl group from an acyl-CoA donor to an amine acceptor, and a member of the M20 peptidase/carboxypeptidase G2 family. We determined that the member of the GNAT superfamily is succinyl-CoA:D-amino acid N-succinyltransferase, the member of the enolase superfamily is N-succinylamino acid racemase (NSAR), and the member of the M20 peptidase/carboxypeptidase G2 family is N-succinyl-L-amino acid hydrolase. We conclude that (1) these enzymes constitute a novel, irreversible pathway for the conversion of D- to L-amino acids and (2) the NSAR reaction is a new physiological function in the MLE subgroup. The NSAR is also functionally promiscuous and catalyzes an efficient OSBS reaction; intriguingly, the operon for menaquinone biosynthesis in G. kaustophilus does not encode an OSBS, raising the possibility that the NSAR is a bifunctional enzyme rather than an accidentally promiscuous enzyme.


The risk of ischemic events increased in patients with asymptomatic carotid stenosis with decreased cerebrovascular reserve.
Identifying high-risk patients with asymptomatic carotid stenosis (ACS) is necessary regardless of whether intensive medical therapy or aggressive treatment is applied. In order to assess the relationship between cerebrovascular reserve (CVR) measured by perfusion CT with inhalation of CO2 and the risk of ischemic events in ACS, this long-term follow-up study was conducted. Forty-five patients with ACS who underwent the examination of CVR measured by perfusion CT with inhalation of CO2 were collected and followed-up for at least 5 years. The primary end point was the occurrence of ipsilateral cerebral ischemic events. HRs and their 95% CI were calculated by Kaplan-Meier survival analysis and Cox regression models. The mean follow-up time was 68.7±10.7 months (40.0-84.0 months). 13 (28.9%) ipsilateral ischemic events were observed. The annual risk of ipsilateral ischemic events was 4.8%. Kaplan-Meier survival analysis and univariate Cox regression analysis indicated that patients with less CVR experienced more ischemic events (p=0.006 and p=0.013, respectively), which was confirmed by multiple Cox regression analysis (p=0.012). CVR measured by perfusion CT may potentially be the factor which can predict the risk of ipsilateral ischemic events in patients with ACS. Multidisciplinary management is necessary for these high-risk patients.


Interferon lambda 3/4 polymorphisms are associated with AIDS-related Kaposi's sarcoma.
Kaposi's sarcoma, the most common AIDS-related cancer, represents a major public concern in resource-limited countries. Single nucleotide polymorphisms within the Interferon lambda 3/4 region (IFNL3/4) determine the expression, function of IFNL4, and influence the clinical course of an increasing number of viral infections. To analyze whether IFNL3/4 variants are associated with susceptibility to AIDS-related Kaposi's sarcoma among MSM enrolled in the Swiss HIV Cohort Study (SHCS). The risk of developing Kaposi's sarcoma according to the carriage of IFNL3/4 SNPs rs8099917 and rs12980275 and their haplotypic combinations was assessed by using cumulative incidence curves and Cox regression models, accounting for relevant covariables. Kaposi's sarcoma was diagnosed in 221 of 2558 MSM Caucasian SHCS participants. Both rs12980275 and rs8099917 were associated with an increased risk of Kaposi's sarcoma (cumulative incidence 15 versus 10%, P = 0.01 and 16 versus 10%, P = 0.009, respectively). Diplotypes predicted to produce the active P70 form (cumulative incidence 16 versus 10%, P = 0.01) but not the less active S70 (cumulative incidence 11 versus 10%, P = 0.7) form of IFNL4 were associated with an increased risk of Kaposi's sarcoma, compared with those predicted not to produce IFNL4. The associations remained significant in a multivariate Cox regression model after adjustment for age at infection, combination antiretroviral therapy, median CD4+ T-cell count nadir and CD4+ slopes (hazard ratio 1.42, 95% confidence interval 1.06-1.89, P = 0.02 for IFLN P70 versus no IFNL4). This study reports for the first time an association between IFNL3/4 polymorphisms and susceptibility to AIDS-related Kaposi's sarcoma.


Sesamin ameliorates hepatic steatosis and inflammation in rats on a high-fat diet via LXRα and PPARα.
Nonalcoholic fatty liver disease (NAFLD) is defined by a nonalcohol relevant pathological accumulation of fat in the liver. Previous studies have shown that sesamin exerts antioxidant effects and improves lipid metabolism of the fatty liver. In this study, we hypothesized that sesamin improves lipid homeostasis of Sprague-Dawley rats fed a high-fat diet (HFD) by regulating the expression of genes related to de novo lipogenesis and β-oxidation. We induced NAFLD in rats with HFD and examined the effect of sesamin in vivo. The results showed that HFD rats accumulated total cholesterol and triacylglycerols in the liver and developed inflammation, as evidenced by the elevation of interleukin-6 and tumor necrosis factor-α in the liver and serum. Sesamin attenuated the disease progression by improving the blood lipid profile in a dose-dependent manner. Sesamin reduced the serum levels of total cholesterol, triacylglycerols, low-density lipoprotein cholesterol, and free fatty acid, whereas it increased the level of high-density lipoprotein cholesterol. Meanwhile, sesamin increased the activities of hepatic glutathione peroxidase and superoxide dismutase while reducing the level of malonaldehyde and cytochrome P450 2E1. Furthermore, higher doses of sesamin reduced the expression of liver X receptor α and its downstream target genes, whereas it upregulated the peroxisome proliferator-activated receptor α-mediated signaling. These findings suggest that sesamin attenuates diet-induced dyslipidemia and inflammation of NAFLD in rats via mechanisms regulated by liver X receptor α and peroxisome proliferator-activated receptor α.


Defects of protein production in erythroid cells revealed in a zebrafish Diamond-Blackfan anemia model for mutation in RPS19.
Diamond-Blackfan anemia (DBA) is a rare congenital red cell aplasia that classically presents during early infancy in DBA patients. Approximately, 25% of patients carry a mutation in the ribosomal protein (RP) S19 gene; mutations in RPS24, RPS17, RPL35A, RPL11, and RPL5 have been reported. How ribosome protein deficiency causes defects specifically to red blood cells in DBA has not been well elucidated. To genetically model the predominant ribosome defect in DBA, we generated an rps19 null mutant through the use of TALEN-mediated gene targeting in zebrafish. Molecular characterization of this mutant line demonstrated that rps19 deficiency reproduced the erythroid defects of DBA, including a lack of mature red blood cells and p53 activation. Notably, we found that rps19 mutants' production of globin proteins was significantly inhibited; however, globin transcript level was either increased or unaffected in rps19 mutant embryos. This dissociation of RNA/protein levels of globin genes was confirmed in another zebrafish DBA model with defects in rpl11. Using transgenic zebrafish with specific expression of mCherry in erythroid cells, we showed that protein production in erythroid cells was decreased when either rps19 or rpl11 was mutated. L-Leucine treatment alleviated the defects of protein production in erythroid cells and partially rescued the anemic phenotype in both rps19 and rpl11 mutants. Analysis of this model suggests that the decreased protein production in erythroid cells likely contributes to the blood-specific phenotype of DBA. Furthermore, the newly generated rps19 zebrafish mutant should serve as a useful animal model to study DBA. Our in vivo findings may provide clues for the future therapy strategy for DBA.


Ultrasensitive fluorescence immunoassay of pepsinogen I based on enzyme-triggered decomposition of AuNCs/MnO2.
Gastric cancer is a prevalent malignant tumor of the gastrointestinal tract accompanied by a high mortality rate; therefore, early gastric cancer screening is critical for improving patient survival. In this study, we present a facile fluorescence immunoassay for highly sensitive screening of pepsinogen I (PG I) based on a one-pot biomimetic mineralization process for the synthesis of gold nanocluster-anchored manganese dioxide (AuNCs/MnO2) nanosheets. MnO2 first quenches the fluorescence of AuNCs through the Förster resonance energy transfer effect, whereas the introduction of ascorbic acid (AA) leads to the decomposition of MnO2 and rapidly recovers the fluorescence of AuNCs. Based on the above principles and phenomena, we developed a sensitive fluorescence immunoassay for the in situ generation of AA to detect PG I. Specifically, in the presence of PG I, the sandwich-type immunoreactivity-enriched alkaline phosphatase-labeled secondary antibody catalyzes the production of AA from the substrate, which enhances the fluorescence intensity. Under optimized conditions, the fluorescence intensity increased linearly with the concentration of PG I (0.05 to 200 ng mL-1) with a limit of detection (LOD) of 0.013 ng mL-1 (S/N = 3). The designed sensing platform has good stability (more than one year) and excellent anti-interference capability and demonstrates satisfactory accuracy for detection in real samples compared to commercial ELISA kits.


Postoperative drift in patients with thyroid ophthalmopathy undergoing unilateral inferior rectus muscle recession.
Extraocular muscles of patients with thyroid ophthalmopathy (TO) may respond differently to strabismus surgery than those of other strabismic patients. This study reports postoperative alignment changes in patients with TO compared with patients with non-restrictive strabismus following unilateral inferior rectus muscle recession (IRR). We reviewed records of patients with and without TO who underwent unilateral IRR. Group A had adjustable muscle sutures, while Group B had permanent or semi-adjustable sutures. Controls were patients undergoing adjustable unilateral IRR for other indications. Mean preoperative hypotropias were 17 ± 9, 21 ± 7, and 11 ± 4 PD for groups A (n=13), B (n=14), and controls (n=19), respectively. Postoperative day one (POD1) measurements after adjustment were 1.2 ± 2.5, 3.7 ± 4.9, and 0.3 ± 2.4 PD, respectively, representing overall undercorrections in all cases (the preoperative deviation was given a positive (+) value and overcorrections were deemed negative (-) deviations). Dose response from linear regression analysis of thyroid patients compared with control patients for IRR was 3.26 PD/mm (SE 0.18) vs 2.38 PD/mm (SE 0.18) (p=0.001). Mean final measurements were -0.7 ± 5.6 (overcorrection), 2.7 ± 5.7, and 1.7 ± 5.7 PD of hypotropia, respectively. Final overcorrections occurred in 23%, 14%, and 16% of patients, for adjustables, permanent sutures, and control subjects, respectively. Drifts from POD1 measurements after adjustment to final measurements were -1.9 ± 4.3, -1.0 ± 4.6, and 1.4 ± 5.9 PD respectively (p=0.05 for comparison between Group A and controls). TO patients with adjustable sutures drift toward postoperative overcorrection.


Cardiopulmonary arrest owing to oesophageal achalasia recovered completely with cardiopulmonary resuscitation followed by therapeutic hypothermia.
We report the first case of cardiopulmonary arrest (CPA), caused by oesophageal achalasia, which recovered completely with cardiopulmonary resuscitation (CPR) followed by therapeutic hypothermia. A 53-year-old woman arrived at our hospital with recovery of spontaneous circulation (ROSC) after cardiac arrest. Dysphagia, vomiting and general fatigue had progressed for a week before. After an ambulance was called for severe dyspnoea, she collapsed in CPA. Emergency medical technicians arrived and CPR was started immediately. She experienced CPA and ROSC twice during transport to the hospital. On arrival, the patient was in respiratory distress prompting immediate intubation to eliminate airway obstruction. A CT scan revealed a transformed, occluded trachea owing to a dilated oesophagus. A large amount of food, air and saliva was suctioned with a nasogastric tube, and the patient was admitted to the intensive care unit for therapeutic hypothermia. Neurological recovery was evident. On the 11th day, pneumatic dilatation was performed and she was discharged on the 33rd day.


Tissue-specific FAH deficiency alters sleep-wake patterns and results in chronic tyrosinemia in mice.
Fumarylacetoacetate hydrolase (FAH) is the last enzyme in tyrosine catabolism, and mutations in the FAH gene are associated with hereditary tyrosinemia type I (HT1 or TYRSN1) in humans. In a behavioral screen of N-ethyl-N-nitrosourea mutagenized mice we identified a mutant line which we named "swingshift" (swst, MGI:3611216) with a nonsynonymous point mutation (N68S) in Fah that caused age-dependent disruption of sleep-wake patterns. Mice homozygous for the mutation had an earlier onset of activity (several hours before lights off) and a reduction in total activity and body weight when compared with wild-type or heterozygous mice. Despite abnormal behavioral entrainment to light-dark cycles, there were no differences in the period or phase of the central clock in mutant mice, indicating a defect downstream of the suprachiasmatic nucleus. Interestingly, these behavioral phenotypes became milder as the mice grew older and were completely rescued by the administration of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione], an inhibitor of 4-hydroxyphenylpyruvate dioxygenase, which is upstream of FAH. Mechanistically, the swst mutation had no effect on the enzymatic activity of FAH, but rather promoted the degradation of the mutant protein. This led to reduced FAH protein levels and enzymatic activity in the liver and kidney (but not the brain or fibroblasts) of homozygous mice. In addition, plasma tyrosine-but not methionine, phenylalanine, or succinylacetone-increased in homozygous mice, suggesting that swst mutants provide a model of mild, chronic HT1.


Trachyonychia associated with alopecia areata: a clinical and pathologic study.
Forty of 1095 patients (3.65%) with alopecia areata had severe nail changes that fulfilled the clinical criteria for the diagnosis of trachyonychia. Twelve of these patients had a nail biopsy. A mild to moderately dense lymphocytic infiltrate associated with exocytosis and spongiosis was detected in the proximal nailfold, nail matrix, nail bed, and hyponychium of 11 patients. One patient showed the pathologic changes of lichen planus; lichen planus of the skin developed 6 months after the nail biopsy. Immunohistochemical characterization on paraffin-embedded sections showed that the inflammatory infiltrate consisted of peripheral T lymphocytes. Immunophenotyping on frozen sections was performed in four cases. The results revealed a T4/T8 ratio of 2:1 and the presence of Langerhans cells in the nail matrix. Our results show that trachyonychia is an uncommon nail manifestation of alopecia areata. Distinctive pathologic features of mild to moderately dense lymphocytic infiltrate associated with exocytosis and spongiosis characterize trachyonychia as well as the other nail abnormalities caused by alopecia areata. The clinical association of trachyonychia with alopecia areata does not exclude that the nail abnormality can be due to other diseases such as lichen planus.


Morphological grayscale reconstruction in image analysis: applications and efficient algorithms.
Two different formal definitions of gray-scale reconstruction are presented. The use of gray-scale reconstruction in various image processing applications discussed to illustrate the usefulness of this transformation for image filtering and segmentation tasks. The standard parallel and sequential approaches to reconstruction are reviewed. It is shown that their common drawback is their inefficiency on conventional computers. To improve this situation, an algorithm that is based on the notion of regional maxima and makes use of breadth-first image scannings implemented using a queue of pixels is introduced. Its combination with the sequential technique results in a hybrid gray-scale reconstruction algorithm which is an order of magnitude faster than any previously known algorithm.


A role for huntington disease protein in dendritic RNA granules.
Regulated transport and local translation of mRNA in neurons are critical for modulating synaptic strength, maintaining proper neural circuitry, and establishing long term memory. Neuronal RNA granules are ribonucleoprotein particles that serve to transport mRNA along microtubules and control local protein synthesis in response to synaptic activity. Studies suggest that neuronal RNA granules share similar structures and functions with somatic P-bodies. We recently reported that the Huntington disease protein huntingtin (Htt) associates with Argonaute (Ago) and localizes to cytoplasmic P-bodies, which serve as sites of mRNA storage, degradation, and small RNA-mediated gene silencing. Here we report that wild-type Htt associates with Ago2 and components of neuronal granules and co-traffics with mRNA in dendrites. Htt was found to co-localize with RNA containing the 3'-untranslated region sequence of known dendritically targeted mRNAs. Knockdown of Htt in neurons caused altered localization of mRNA. When tethered to a reporter construct, Htt down-regulated reporter gene expression in a manner dependent on Ago2, suggesting that Htt may function to repress translation of mRNAs during transport in neuronal granules.


Effects of chloramphenicol on the mitochondrial respiratory chain in the wild strain and in a cytoplasmic chloramphenicol-resistant mutant of Tetrahymena pyriformis.
The effects of chloramphenicol (CAP) on mitochondrial respiratory activity in the wild strain (ST) and in a cytoplasmic CAP-resistant mutant (STR1) of Tetrahymena pyriformis were studied by determining oxygen consumption, by spectrophotometry, and by cytochemistry. In the absence of CAP both strains had the same respiration capacity, and the low-temperature spectra of their isolated mitochondria were similar. Furthermore, the mitochondria of both strains showed a positive reaction with diaminobenzidine, denoting a similar cytochrome oxidase activity. However, when cells were grown in CAP for 24 or 48 h, the peaks of cytochrome oxidase and cytochromb b were almost absent in the wild type. In this type the oxygen consumption was greatly decreased, and the mitochondria were no longer stained by diaminobenzidine. In the mutant, the peaks of cytochrome oxidase and cytochrome b were decreased only; respiration was less affected than in the wild type, and cytochrome oxidase activity was still disclosed by the diaminobenzidine reaction. These results show that CAP inhibits the synthesis of two cytochromes (b and oxidase) which are partially translated into the mitochrondria of T. pyriformis. In the mutant, CAP reduces only the mitochondrial translation, resulting in reduced mitochondrial activity and reduced growth rate of the cell. These results are compared with the nucleo-mitochondrial regulation mechanisms discussed in our previous works.


Attenuating Cholinergic Transmission Increases the Number of Satellite Cells and Preserves Muscle Mass in Old Age.
In addition to driving contraction of skeletal muscles, acetylcholine (ACh) acts as an anti-synaptogenic agent at neuromuscular junctions (NMJs). Previous studies suggest that aging is accompanied by increases in cholinergic activity at the NMJ, which may play a role in neuromuscular degeneration. In this study, we hypothesized that moderately and chronically reducing ACh could attenuate the deleterious effects of aging on NMJs and skeletal muscles. To test this hypothesis, we analyzed NMJs and muscle fibers from heterozygous transgenic mice with reduced expression of the vesicular ACh transporter (VAChT; VKDHet), which present with approximately 30% less synaptic ACh compared to control mice. Because ACh is constitutively decreased in VKDHet, we first analyzed developing NMJs and muscle fibers. We found no obvious morphological or molecular differences between NMJs and muscle fibers of VKDHet and control mice during development. In contrast, we found that moderately reducing ACh has various effects on adult NMJs and muscle fibers. VKDHet mice have significantly larger NMJs and muscle fibers compared to age-matched control mice. They also present with reduced expression of the pro-atrophy gene, Foxo1, and have more satellite cells in skeletal muscles. These molecular and cellular features may partially explain the increased size of NMJs and muscle fibers. Thus, moderately reducing ACh may be a therapeutic strategy to prevent the loss of skeletal muscle mass that occurs with advancing age.


Translation and validation of the Greek "Minnesota Living with Heart Failure" questionnaire.
The Minnesota Living with Heart Failure Questionnaire (MLHFQ) is an important measurement instrument for assessing the health-related quality of life (HRQOL) among heart failure patients. The purpose of this study was to translate and validate the MLHFQ in the Greek language. Three hundred forty-four consecutive adult patients from three General Hospitals, two in Athens and one in another part of the country, who were diagnosed with chronic heart failure, and 347 healthy controls were enrolled in the study from March 2009 to March 2010. The questionnaire instrument was translated from English, back-translated, and reviewed by a committee of experts. The psychometric measurements that were performed included reliability coefficients and Explanatory Factor Analysis (EFA), using a Varimax rotation and Principal Components Method. In a further step, confirmatory analysis (CFA)--known as structural equation modeling--of the principal components was conducted. The internal consistency of the Greek MLHFQ version was found to be 0.97, using Cronbach's alpha coefficient. An exploratory factor analysis identified two domains that accounted for 72.5% of the variance of MLHFQ items; the area under the ROC curve was calculated at 0.942 and the logistic estimate for the threshold score of 24.50 provided the model with 95.1% sensitivity and 99.8% specificity. Additionally, the CFA demonstrated that the two-factor model offered a very good fit to our data. Our data indicate that the Greek MLHFQ is a reliable and valid tool for assessing HRQOL among patients with heart failure. Health professionals can use it in their clinical practice to improve their evaluation of these patients.


Successful medical management of lumbar intervertebral disc prolapse in a ferret.
Thoracolumbar disc herniation was diagnosed in a two-year-old ferret using a myelogram followed by a computed tomography scan. The ferret was paraplegic with no control over urination and defecation. Conservative treatment that included cage rest and passive range of motion exercises for three weeks followed by extensive physiotherapy and hydrotherapy resulted in full recovery two months after the onset of treatment. Although intervertebral disc disease has been reported previously in four ferrets, this is the first report in which a postmyelogram computed tomography was used to demonstrate the herniated disc, and physiotherapy was used as principal treatment modality.


Duplex Ultrasound Surveillance After Uncomplicated Endovascular Abdominal Aortic Aneurysm Repair.
Health-care costs and risks of radiation and intravenous contrast exposure challenge computed tomography angiography (CTA) as the standard surveillance method after endovascular abdominal aortic aneurysm repair (EVAR). We reviewed our experience using Duplex ultrasound scan (DUS) as an initial and subsequent surveillance technique after uncomplicated EVAR. The medical records of patients who underwent EVAR from 2004 to 2014 with at least 1 postoperative imaging study were retrospectively reviewed. Duplex ultrasound scan was the primary modality, with CTA reserved for patients with suspicious findings. Mean follow-up was 3.2 years for 266 patients. Fifty-seven endoleaks (7 type I, 50 type II) were detected in 51 patients (19%). Nineteen (33%) endoleaks were identified and monitored by DUS alone. Nine (16%) endoleaks were identified on CTA without prior DUS. Twenty-two (39%) endoleaks were identified on DUS and confirmed by CTA; 6 of these patients had a secondary intervention. When compared to subsequent CTA, there were 7 discordant results: 4 false-negative and 3 false-positive endoleaks on DUS. Two of these patients with discordant results required intervention. Follow-up CTA was not obtained for the other 2 patients due to severe comorbidities including renal disease. One of these patients eventually developed abdominal aortic aneurysm rupture and death. Among 88 patients with both DUS and CTA, positive predictive value and negative predictive value for DUS were 0.88 and 0.94, respectively. Sac size on DUS compared to CTA resulted in an interclass correlation coefficient of r = .84. In our experience, DUS was safe and effective for initial and follow-up surveillance after uncomplicated EVAR.


Phase-separated CCER1 coordinates the histone-to-protamine transition and male fertility.
Idiopathic fertility disorders are associated with mutations in various genes. Here, we report that coiled-coil glutamate-rich protein 1 (CCER1), a germline-specific and intrinsically disordered protein (IDP), mediates postmeiotic spermatid differentiation. In contrast, CCER1 deficiency results in defective sperm chromatin compaction and infertility in mice. CCER1 increases transition protein (Tnp1/2) and protamine (Prm1/2) transcription and mediates multiple histone epigenetic modifications during the histone-to-protamine (HTP) transition. Immiscible with heterochromatin in the nucleus, CCER1 self-assembles into a polymer droplet and forms a liquid-liquid phase-separated condensate in the nucleus. Notably, we identified loss-of-function (LoF) variants of human CCER1 (hCCER1) in five patients with nonobstructive azoospermia (NOA) that were absent in 2713 fertile controls. The mutants led to premature termination or frameshift in CCER1 translation, and disrupted condensates in vitro. In conclusion, we propose that nuclear CCER1 is a phase-separated condensate that links histone epigenetic modifications, HTP transitions, chromatin condensation, and male fertility.


Impact of personality traits on learners' navigational behavior patterns in an online course: a lag sequential analysis approach.
Personality is considered as the internal factor that defines a person's behavior. Therefore, providing adaptive features and personalized support in online learning by considering learners' personalities can improve their learning experiences and outcomes. In this context, several research studies have investigated the impact of personality differences in online learning. However, little is known about how personality differences affect learners' behavior while learning. To fill this gap, this study applies a lag sequential analysis (LSA) approach to understand learners' navigational behavior patterns in an online three-months course of 65 learners based on their personalities. In this context, the five factor model (FFM) model was used to identify learners' personalities. The findings revealed that learners with different personalities use different strategies to learn and navigate within the course. For instance, learners high in extraversion tend to be extrinsically motivated. They therefore significantly navigated between viewing the course module and their personal achievements. The findings of this study can contribute to the adaptive learning field by providing insights about which personalization features can help learners with different personalities. The findings can also contribute to the field of automatic modeling of personality by providing information about differences in navigational behavior based on learners' personalities.


The use of modified and non-natural nucleotides provide unique insights into pro-mutagenic replication catalyzed by polymerase eta.
This report evaluates the pro-mutagenic behavior of 8-oxo-guanine (8-oxo-G) by quantifying the ability of high-fidelity and specialized DNA polymerases to incorporate natural and modified nucleotides opposite this lesion. Although high-fidelity DNA polymerases such as pol δ and the bacteriophage T4 DNA polymerase replicating 8-oxo-G in an error-prone manner, they display remarkably low efficiencies for TLS compared to normal DNA synthesis. In contrast, pol η shows a combination of high efficiency and low fidelity when replicating 8-oxo-G. These combined properties are consistent with a pro-mutagenic role for pol η when replicating this DNA lesion. Studies using modified nucleotide analogs show that pol η relies heavily on hydrogen-bonding interactions during translesion DNA synthesis. However, nucleobase modifications such as alkylation to the N2 position of guanine significantly increase error-prone synthesis catalyzed by pol η when replicating 8-oxo-G. Molecular modeling studies demonstrate the existence of a hydrophobic pocket in pol η that participates in the increased utilization of certain hydrophobic nucleotides. A model is proposed for enhanced pro-mutagenic replication catalyzed by pol η that couples efficient incorporation of damaged nucleotides opposite oxidized DNA lesions created by reactive oxygen species. The biological implications of this model toward increasing mutagenic events in lung cancer are discussed.


Posterosuperior rotator cuff tears: classification, pattern recognition, and treatment.
The posterosuperior rotator cuff, composed of the supraspinatus and infraspinatus tendons, is the most common site for full-thickness rotator cuff tears and represents a significant source of shoulder disability worldwide. Recognition of and classification of full-thickness tear patterns are essential in order to optimize surgical treatment and to improve prognosis. Until recently, tear patterns have been described using one- or two-dimensional classification systems. Three-dimensional pattern recognition is critical to achieving the most successful outcome possible. For more complex patterns, a combination of side-to-side stitching, margin convergence, and interval slide techniques may be needed to achieve a tension-free tendon-bone repair. Biomechanical and anatomic evidence supports the use of linked double-row repairs for most full-thickness tears. Although double-row repairs seem to result in improved structural outcomes, clinical evidence has not shown differences in outcomes scores between single-row and double-row repairs. Single-row repair may be performed in partial-thickness, small full-thickness, or very massive, immobile tears, whereas double-row repair may be performed in most other cases.


A tonal-language benefit for pitch in normally-hearing and cochlear-implanted children.
In tonal languages, voice pitch inflections change the meaning of words, such that the brain processes pitch not merely as an acoustic characterization of sound but as semantic information. In normally-hearing (NH) adults, this linguistic pressure on pitch appears to sharpen its neural encoding and can lead to perceptual benefits, depending on the task relevance, potentially generalizing outside of the speech domain. In children, however, linguistic systems are still malleable, meaning that their encoding of voice pitch information might not receive as much neural specialization but might generalize more easily to ecologically irrelevant pitch contours. This would seem particularly true for early-deafened children wearing a cochlear implant (CI), who must exhibit great adaptability to unfamiliar sounds as their sense of pitch is severely degraded. Here, we provide the first demonstration of a tonal language benefit in dynamic pitch sensitivity among NH children (using both a sweep discrimination and labelling task) which extends partially to children with CI (i.e., in the labelling task only). Strong age effects suggest that sensitivity to pitch contours reaches adult-like levels early in tonal language speakers (possibly before 6 years of age) but continues to develop in non-tonal language speakers well into the teenage years. Overall, we conclude that language-dependent neuroplasticity can enhance behavioral sensitivity to dynamic pitch, even in extreme cases of auditory degradation, but it is most easily observable early in life.


An Exploration of Mineral Density, Elemental and Chemical Composition of Primary Teeth in Relation to Cord-Blood Vitamin D, Using Laboratory Analysis Techniques.
Postnatally, severe vitamin D deficiency commonly results in rickets as well as potential defects in tooth mineralization. The effects of milder deficiency on oral health outcomes later in life are still unclear. This study used micro-computed tomography (μCT), energy dispersive X-ray analysis (EDX), and Raman spectroscopy to investigate mineral density, total density, and elemental composition of enamel and dentine in 63 exfoliated primary incisors from participants with known 25-hydroxyvitamin D levels (25-OHD) at birth. No differences in mineralization and chemical composition using μCT and EDX analysis were observed irrespective of 25-OHD status. Subtle structural differences were observed via Raman spectroscopy, with more crystalline enamel observed in those with sufficient 25-OHD at birth. Although subtle, the differences seen suggest further attention should be given to children with known milder levels of vitamin D deficiency in early life. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).


Safety of bevacizumab in metastatic breast cancer patients undergoing surgery.
Evaluate the safety of surgery in relation to bevacizumab in the first-line treatment of metastatic breast cancer (mBC) in two international trials. The incidence, type and timing of post-surgical bleeding events and wound-healing complications were assessed in surgical patients in the AVastin And DOcetaxel (AVADO) (NCT00333775) and Avastin THErapy for advaNced breAst cancer (ATHENA) (NCT00448591) trials. Both study protocols followed recommendations to withhold bevacizumab for at least 6 weeks before elective surgery and to wait 28 days (or until the wound was fully healed) after major surgery before recommencing bevacizumab therapy. In AVADO, 221 surgical procedures (55 major, 166 minor) were performed in 155 patients. In ATHENA, 1190 surgical procedures (435 major, 755 minor) were performed in 672 patients. One bevacizumab-treated AVADO patient (0.9%) who underwent surgery experienced a grade 3 bleeding event. In ATHENA, six patients (0.9%) who underwent surgery experienced grade 3 bleeding events and one patient (0.1%) experienced a grade 4 bleeding event. No grade 5 bleeding events in patients undergoing surgery were reported in either study. One grade 3 wound-healing complication was reported in each of the AVADO arms: placebo (n=46, 2.2%), bevacizumab 7.5mg/kg (n=57, 1.8%) and bevacizumab 15mg/kg (n=52, 1.9%). Incidence of grade 3-4 wound-healing complications in ATHENA was 2.2% and 1.3% in patients undergoing minor or major surgery, respectively. Surgery can be performed on patients with mBC undergoing bevacizumab therapy with a low risk of severe bleeding or wound-healing complications post surgery, if current labelling recommendations are adhered to.


Deciphering the Mechanism of Glyphosate Resistance in Amaranthus palmeri by Cytogenomics.
In agriculture, various chemicals are used to control the weeds. Out of which, glyphosate is an important herbicide invariably used in the cultivation of glyphosate-resistant crops to control weeds. Overuse of glyphosate results in the evolution of glyphosate-resistant weeds. Evolution of glyphosate resistance (GR) in Amaranthus palmeri (AP) is a serious concern in the USA. Investigation of the mechanism of GR in AP identified different resistance mechanisms of which 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene amplification is predominant. Molecular analysis of GR AP identified the presence of a 5- to >160-fold increase in copies of the EPSPS gene than in a glyphosate-susceptible (GS) population. This increased copy number of the EPSPS gene increased the genome size ranging from 3.5 to 11.8%, depending on the copy number compared to the genome size of GS AP. FISH analysis using a 399-kb EPSPS cassette derived from bacterial artificial chromosomes (BACs) as probes identified that amplified EPSPS copies in GR AP exist in extrachromosomal circular DNA (eccDNA) in addition to the native copy in the chromosome. The EPSPS gene-containing eccDNA having a size of ∼400 kb is termed EPSPS-eccDNA and showed somatic mosacism in size and copy number. EPSPS-eccDNA has a genetic mechanism to tether randomly to mitotic or meiotic chromosomes during cell division or gamete formation and is inherited to daughter cells or progeny generating copy number variation. These eccDNAs are stable genetic elements that can replicate and exist independently. The genomic characterization of the EPSPS locus, along with the flanking regions, identified the presence of a complex array of repeats and mobile genetic elements. The cytogenomics approach in understanding the biology of EPSPS-eccDNA sheds light on various characteristics of EPSPS-eccDNA that favor GR in AP.


Chitosan-lipid nanoparticles (CS-LNPs): Application to siRNA delivery.
To benefit from the biocompatibility of lipid nanoparticles associated with the transfection ability of chitosan, small chitosan lipid nanoparticles (CS-LNPs) dedicated to SiRNA delivery were formulated by an easy-to-implement one-step process. Formulations of CS-LNPs (lipid core stabilized by a shell comprising phospholipids/cationic lipids and hydrophobically modified chitosan) were optimized for their physico-chemical properties (size, zeta potential, colloidal stability) according to their shell composition. In particular, amphiphilic chitosan with various molecular weight and C12 degrees of substitution, and different phospholipids and cationic lipids (lecithin, DOTAP, DOPE) were included at the particle surface at different ratios. The ability of the particles for SiRNA complexation, NIH3T3 cell transfection, and ERK1 downregulation, were studied. Lipid nanoparticles formulated with 15,000g/mol 2% C12 substituted chitosan, DOTAP and DOPE, mediated 40% ERK1 downregulation efficiency, comparable to lipofectamine™ RNAimax, while displaying no cytotoxicity up to 500μg/mL.


The effects of general anaesthesia on antroduodenal motility, gastric pH and gastric emptying in man.
This survey was based on five recently published papers. The aims of the studies were to to investigate the effect of general anaesthesia on antroduodenal motility, gastric pH and gastric emptying. Antroduodenal motility was measured by manometry, gastric pH by a stomach microelectrode and gastric emptying rate using paracetamol absorption as an index. Two studies deal with antroduodenal motility during basal conditions and after administration of paracetamol and diazepam in 11 healthy volunteers. Three studies deal with the effects of 4 different methods of general anaesthesia on antroduodenal motility, gastric pH and gastric emptying rate in 43 patients undergoing orthopedic or plastic surgery not involving the abdomen having four different methods of general anaesthesia. In the healthy volunteers a close connection between antral activity and gastric emptying rate was found. Paracetamol had no effects on gastric pH. The changes in motility (a shortening of phase III and a decline in the incidence of antral phase III activity) were minor and ascribed to diurnal variations and by a possible stimulation of antral activity by the positioning of the manometric tube. It was found that no paracetamol was absorbed in the quiescent phase I, irrespective of its duration, and that the absorption rate of paracetamol correlated with the duration of phase II and the occurrence of antral activity during phases II and III. The effects of premedication with diazepam on antroduodenal motility, gastric pH and gastric emptying rate was investigated in the same healthy volunteers prior to the patient studies. Also in this study a correlation between antral activity and gastric emptying rate was found. Furthermore, it was observed that some volunteers had fast gastric emptying rate, indicated by fast absorption of paracetamol, the first study day and slow the second. It seemed that any intra- or interindividual variation in gastric emptying rate arises from individual variations in antroduodenal motility. Diazepam tended to increase gastric emptying rate by enhancing the amplitudes of antral contractions and the motility index during phase II and pH of the gastric juice increased. Gastroduodenal motility was found to be normal in patients awaiting elective orthopedic or plastic surgery when premedicated with diazepam. pH of the gastric contents was not different from the findings in volunteers after administration of diazepam. All methods of general anaesthesia reduced the duration of the interdigestive motility complex, mainly by a reduction of phase II. General anaesthesia with halothane and enflurane depressed antral motility.(ABSTRACT TRUNCATED AT 400 WORDS)


Motor control in basal ganglia circuits using fMRI and brain atlas approaches.
In this study, we examined how the motor, premotor and associative basal ganglia territories process movement parameters such as the complexity and the frequency of movement. Twelve right-handed volunteers were studied using EPI BOLD contrast (3 T) while performing audio-paced finger tapping tasks designed to differentiate basal ganglia territories. Tasks varied movement complexity (repetitive index tapping, simple sequence of finger movements and complex sequence of 10 moves) and frequency (from 0.5 to 3 Hz). Activation maps were coregistered onto a 3-D brain atlas derived from post-mortem brains. Three main patterns of activation were observed. In the posterior putamen and the sensorimotor cortex, signal increased with movement frequency but not with movement complexity. In premotor areas, the anterior putamen and the ventral posterolateral thalamus, signal increased regularly with increasing movement frequency and complexity. In rostral frontal areas, the caudate nucleus, the subthalamic nucleus and the ventral anterior/ventrolateral thalamus, signal increased mainly during the complex task and the high frequency task (3 Hz). These data show the different roles of motor, premotor and associative basal ganglia circuits in the processing of motor-related operations and suggest that activation can be precisely located within the entire circuitry of the basal ganglia.


A fluorescence perspective on the differential interaction of riboflavin and flavin adenine dinucleotide with cucurbit[7]uril.
The interaction of the macrocyclic host, cucurbit[7]uril (CB7), with riboflavin (RF) and its derivative, flavin adenine dinucleotide (FAD), has been investigated using absorption and steady-state and time-resolved fluorescence measurements. Interestingly, in the presence of CB7, the fluorescence intensity of RF is quenched, whereas the fluorescence intensity of FAD is enhanced. It is proposed that the fluorescence quenching of RF results from the tautomerization of its isoalloxazine moiety from the lactam to the lactim forms, upon binding to CB7. Such a tautomerization can be brought about since the two lactim forms have higher dipole moments than the lactam form of RF, and thus experience much stronger dipole-dipole interactions (and hence greater binding affinities) with CB7 in the former cases than in the latter. This tautomerization in the presence of CB7 leads to a significant reduction in the observed radiative decay rate and hence a reduction in the fluorescence intensity of RF. Binding of CB7 with RF is confirmed by an increase in the rotational correlation time of RF in the presence of CB7. Geometry optimization studies indicate the formation of an exclusion complex between CB7 and RF, possibly stabilized by H-bonding interactions, as also suggested by the characteristic red shift in the absorption spectra of the CB7-RF system. In the case of FAD, both the isoalloxazine ring and the adenine moiety can interact with the CB7 host. In aqueous solutions, a good fraction of FAD molecules exists in a "closed" conformation with the adenine and isoalloxazine rings stacked together, thus leading to very efficient fluorescence quenching due to the ultrafast intramolecular electron transfer from adenine to the isoalloxazine moiety. Binding of the adenine and/or the isoalloxazine moiety of FAD with CB7 inhibits the stacking interaction and changes the "closed" conformation to the "open" conformation, wherein the adenine and isoalloxazine moieties are widely separated, thus prohibiting the electron transfer process. This reduces the inherent fluorescence quenching of FAD molecule and results in the observed fluorescence enhancement. As observed for RF, the interaction of CB7 with the isoalloxazine ring of FAD should cause fluorescence quenching due to the lactam to lactim tautomerization process. However, in the interplay between the above two opposing effects, the fluorescence enhancement due to the modulation in the conformational dynamics of FAD by the CB7 host predominates. The conformational change is in fact supported by the observation of a long lifetime component in the fluorescence decay of FAD in the presence of CB7. Moreover, at acidic pH, when FAD is already present mainly in the "open" form, the conformational dynamics no longer plays any major role and the fluorescence of FAD is quenched by CB7, as expected, due to the tautomerization at the isoalloxazine moiety.


Targeting cell cycle machinery as a molecular mechanism of sulforaphane in prostate cancer prevention.
Epidemiological studies recently concluded that consumption of cruciferous vegetables such as broccoli, cabbage, and cauliflower, etc. is inversely related to prostate cancer risk, although the mechanism of prevention and the responsible phytochemicals are unknown. Since clinically significant prostate cancer eventually can grow independent of androgen, the association of the growth and tumorigenesis of such prostate cancer cells with sulforaphane (SFN) which is a predominant isothiocyanate in cruciferous vegetables, investigated. These vegetables contain high concentrations of glucosinolate glucoraphanin, which yield sulforaphane when hydrolyzed by the plant enzyme myrosinase. This study showed that exposure of human androgen-independent DU-145 prostate cancer cells to SFN resulted in the inhibition of growth and tumorigenesis, as revealed by a reduction in cell density, DNA synthesis, and clonogenesis. Analyses of the mechanism revealed that SFN mediated cell cycle arrest by modulating the expression and functions of cell cycle regulators. SFN induced signals that inhibited the activity of cyclin-dependent kinase cdk4 with an up-stream induction of cdk inhibitor p21WAF-1/Cip-1, and reduced cyclin D1. The inhibition of cdk kinase activity could be affected with <1 micro M SFN within 24 h. As a result, phosphorylation of Rb proteins, which activates the transition from G1- to S-phase, was significantly decreased and the cell cycle progression retarded. SFN also down-regulated the expression of bcl-2, a suppressor of apoptosis, and activated caspases to execute apoptosis in the prostate cancer cells. The regulators of cell cycle have thus been revealed as targets of sulforaphane for growth arrest and apoptosis induction. The potential of SFN, as an active dietary factor to inhibit initiation and post-initiation of prostate cancer carcinogenesis is discussed.


The Role of Epithelial to Mesenchymal Transition in Human Amniotic Membrane Rupture.
Biochemical weakening of the amnion is a major factor preceding preterm premature rupture of membranes (PPROMs), leading to preterm birth. Activation of matrix metalloproteinases (MMPs) is known to play a key role in collagen degradation of the amnion; however, epithelial to mesenchymal transition (EMT) that is also induced by MMP activation has not been investigated as a mechanism for amnion weakening. To measure amniotic EMT associated with vaginal delivery (VD) compared with unlabored cesarean sections (CSs), and to assess changes in amniotic mechanical strength with pharmacologic inhibitors and inducers of EMT, thus testing the hypothesis that EMT is a key biochemical event that promotes amniotic rupture. (1) Amnions taken from VD contained a significantly increased number of mesenchymal cells relative to epithelial cells compared with unlabored CS by fluorescence-activated cell sorting analysis (60% vs 10%); (2) tumor necrosis factor (TNF)-α stimulation of amniotic epithelial cells increased expression of the mesenchymal marker vimentin after 2 days; (3) EMT inhibitor, etodolac, significantly increased the time and mechanical pressure required to rupture the amnion; and (4) TNF-α and another pharmacologic EMT inducer, ethacridine, decreased the time and mechanical pressure required for amnion rupture, further confirming that the mesenchymal phenotype significantly weakens the amnion. This work demonstrated amniotic cell EMT was associated with labor and EMT decreased the tensile strength of the amnion. These findings suggest a role for EMT in the pathophysiology of PPROM and may provide a basis for development of therapies to prevent preterm labor.


Ostomy Usage for Colorectal Trauma in Combat Casualties.
The role for diverting ostomy as a method to help reduce morbidity and mortality has been well established in the combat trauma population. However, factors that influence the type of ostomy used and which ostomies become permanent are poorly studied. We examine patterns of ostomy usage and reversal in a large series of combat trauma patients. We performed a retrospective review of combat casualties treated at our continental U.S. military treatment facility from 2003 to 2015. All patients who underwent ostomy formation were included. Clinical and demographic factors were collected for all patients including the type of ostomy and whether or not ostomy reversal took place. Patients were grouped and analyzed based on ostomy type and by ostomy reversal. We identified 202 patients who had ostomies created. End colostomies were most common (N = 149) followed by loop colostomies (N = 34) and end ileostomies (N = 19). Casualties that underwent damage control laparotomy (DCL) were less likely to have a loop colostomy created (p < 0.001). Ostomy reversal occurred in 89.9% of patients. There was no difference in ostomy reversal rates by ostomy type (p = 0.080). Presence of a pelvic fracture was associated with permanent ostomy (OR = 3.28, p = 0.019), but no factor independently predicted a permanent ostomy on multivariate analysis. DCL and a severe perineal injury most strongly influence ostomy type selection. Most patients undergo colostomy reversal, and no factor independently predicted an ostomy being permanent. These findings provide a framework for understanding the issue of fecal diversion in the combat trauma population and inform military surgeons about injury patterns and treatment options.


Association between the Serum Sodium Levels and the Response to Tolvaptan in Liver Cirrhosis Patients with Ascites and Hyponatremia.
Objective Hyponatremia is closely associated with the pathophysiology of cirrhosis. However, the association between the serum sodium level and the response to tolvaptan is unclear. This study evaluated the factors related to the tolvaptan response and the prognosis in cirrhosis patients with ascites and hyponatremia. Methods We retrospectively reviewed the clinical records of cirrhosis patients hospitalized for treatment with tolvaptan. The associations of patient baseline characteristics with the tolvaptan response after one week and of the characteristics after one-month tolvaptan treatment with the prognosis were analyzed. Results We analyzed 83 cirrhosis patients with ascites, including 34 patients with hyponatremia. The response rates to tolvaptan in patients with serum sodium <130 mEq/L, 130-135 mEq/L, and >135 mEq/L were 20%, 66%, and 58%, respectively (p=0.22). The serum sodium level was associated with the response to tolvaptan [odds ratio=1.18; 95% confidence interval (CI) =1.02-1.37; p=0.029]. In patients with hyponatremia, the serum sodium level after 1-month tolvaptan treatment was increased compared to baseline (132 mEq/L vs. 136 mEq/L, p=0.006), and an increasing serum sodium level was associated with a lower risk of mortality (hazard ratio=0.85; 95% CI=0.75-0.97; p=0.016). The survival rate was higher in patients with an increase in the serum sodium level after 1 month than in patients with a decreased serum sodium level (p=0.023). Conclusion Tolvaptan treatment was effective in cirrhosis patients with ascites and hyponatremia, but a low serum sodium level was associated with non-responsiveness to tolvaptan. An increased serum sodium level after one-month tolvaptan treatment may positively influence the mortality risk in cirrhosis patients with hyponatremia.


Parasitic helminth genomics. Filarial Genome Project.
The initiation of genome projects on helminths of medical importance promises to yield new drug targets and vaccine candidates in unprecedented numbers. In order to exploit this emerging data it is essential that the user community is aware of the scope and quality of data available, and that the genome projects provide analyses of the raw data to highlight potential genes of interest. Core bioinformatics support for the parasite genome projects has promoted these approaches. In the Brugia genome project, a combination of expressed sequence tag sequencing from multiple cDNA libraries representing the complete filarial nematode lifecycle, and comparative analysis of the sequence dataset, particularly using the complete genome sequence of the model nematode C. elegans, has proved very effective in gene discovery.


Ghrelin-mediated sympathoinhibition and suppression of inflammation in sepsis.
Sepsis, a systemic inflammatory response to infection, continues to carry a high mortality despite advances in critical care medicine. Elevated sympathetic nerve activity in sepsis has been shown to contribute to early hepatocellular dysfunction and subsequently multiple organ failure, resulting in a poor prognosis, especially in the elderly. Thus, suppression of sympathetic nerve activity represents a novel therapeutic option for sepsis. Ghrelin is a 28-amino acid peptide shown to inhibit sympathetic nerve activity and inflammation in animal models of tissue injury. Age-related ghrelin hyporesponsiveness has also been shown to exacerbate sepsis. However, the mechanistic relationship between ghrelin-mediated sympathoinhibition and suppression of inflammation remains poorly understood. This review assesses the therapeutic potential of ghrelin in sepsis in the context of the neuroanatomical and molecular basis of ghrelin-mediated suppression of inflammation through inhibition of central sympathetic outflow.


Association between maxillary sinus pathologies and healthy teeth.
The proximity of the roots to the maxillary sinus can create a variety of risks. To evaluate the relationship between the roots of healthy teeth and the maxillary sinus, as well as the occurrence of sinus pathologies. Three radiologists analyzed 109 cone beam computed tomography (CBCT) images. The Kappa test was used to assess the intra- and inter-rater agreement. The chi-squared test and prevalence ratio were used to test the hypothesis that roots of healthy teeth in the maxillary sinus favored the occurrence of sinus pathologies (p=0.01). Intra- and inter-rater agreement ranged from good to excellent. The chi-squared test demonstrated a statistically significant difference (p=0.006) between the tooth roots in diseased maxillary sinuses (6.09%) and those in normal sinuses (3.43%). The prevalence ratio test showed a statistically significant higher prevalence of tooth roots in diseased sinuses than in normal sinuses (p<0.0001). Roots in the maxillary sinus were 1.82 times more associated with diseased sinuses. Dental roots in the maxillary sinus are almost twice as likely to be associated with diseased sinuses than normal sinuses. Healthy teeth whose roots are inside the maxillary sinus may induce an inflammatory response in the sinus membrane. It is suspected that dental procedures may exacerbate the condition.


Sonic hedgehog therapy in a mouse model of age-associated impairment of skeletal muscle regeneration.
Sonic hedgehog (Shh) is a morphogen regulating muscle development during embryogenesis. We have shown that the Shh pathway is postnatally recapitulated after injury and during regeneration of the adult skeletal muscle and regulates angiogenesis and myogenesis after muscle injury. Here, we demonstrate that in 18-month-old mice, there is a significant impairment of the upregulation of the Shh pathway that physiologically occurs in the young skeletal muscle after injury. Such impairment is even more pronounced in 24-month-old mice. In old animals, intramuscular therapy with a plasmid encoding the human Shh gene increases the regenerative capacities of the injured muscle, in terms of Myf5-positive cells, regenerating myofibers, and fibrosis. At the molecular level, Shh treatment increases the upregulation of the prototypical growth factors, insulin-like growth factor-1 and vascular endothelial growth factor. These data demonstrate that Shh increases regeneration after injury in the muscle of 24-month-old mice and suggest that the manipulation of the Shh pathway may be useful for the treatment of muscular diseases associated with aging.


Highly Efficient Photothermal Icephobic/de-Icing MOF-Based Micro and Nanostructured Surface.
Photothermal materials have gained considerable attention in the field of anti-/de-icing due to its environmental friendliness and energy saving. However, it is always significantly challenging to obtain solar thermal materials with hierarchical structure and simultaneously demonstrate both the ultra-long icing delay ability and the superior photothermal de-icing ability. Here, a photothermal icephobic MOF-based micro and nanostructure surface (MOF-MNS) is presented, which consists of micron groove structure and fluorinated MOF nanowhiskers. The optimal MOF-M250 NS can achieve solar absorption of over 98% and produce a high temperature increment of 65.5 °C under 1-sun illumination. Such superior photothermal-conversion mechanism of MOF-M250 NS is elucidated in depth. In addition, the MOF-M250 NS generates an ultra-long icing delay time of ≈3960 s at -18 °C without solar illumination, achieving the longest delay time, which isn't reported before. Due to its excellent solar-to-heat conversation ability, accumulated ice and frost on MOF-M250 NS can be rapidly melted within 720 s under 1-sun illumination and it also holds a high de-icing rate of 5.8 kg m-2 h-1 . MOF-M250 NS possesses the versatility of mechanical robustness, chemical stability, and low temperature self-cleaning, which can synergistically reinforce the usage of icephobic surfaces in harsh conditions.


Nonlinear simulation of tumor necrosis, neo-vascularization and tissue invasion via an adaptive finite-element/level-set method.
We present a multi-scale computer simulator of cancer progression at the tumoral level, from avascular stage growth, through the transition from avascular to vascular growth (neo-vascularization), and into the later stages of growth and invasion of normal tissue. We use continuum scale reaction-diffusion equations for the growth component of the model, and a combined continuum-discrete model for the angiogenesis component. We use the level set method for describing complex topological changes observed during growth such as tumor splitting and reconnection, and capture of healthy tissue inside the tumor. We use an adaptive, unstructured finite element mesh that allows for finely resolving important regions of the computational domain such as the necrotic rim, the tumor interface and around the capillary sprouts. We present full nonlinear, two-dimensional simulations, showing the potential of our virtual cancer simulator. We use microphysical parameters characterizing malignant glioma cells, obtained from recent in vitro experiments from our lab and from clinical data, and provide insight into the mechanisms leading to infiltration of the brain by the cancer cells. The results indicate that diffusional instability of tumor mass growth and the complex interplay with the developing neo-vasculature may be powerful mechanisms for tissue invasion.


Exploring Early Pre-Symptomatic Detection of Influenza Using Continuous Monitoring of Advanced Physiological Parameters during a Randomized Controlled Trial.
Early detection of influenza may improve responses against outbreaks. This study was part of a clinical study assessing the efficacy of a novel influenza vaccine, aiming to discover distinct, highly predictive patterns of pre-symptomatic illness based on changes in advanced physiological parameters using a novel wearable sensor. Participants were frequently monitored 24 h before and for nine days after the influenza challenge. Viral load was measured daily, and self-reported symptoms were collected twice a day. The Random Forest classifier model was used to classify the participants based on changes in the measured parameters. A total of 116 participants with ~3,400,000 data points were included. Changes in parameters were detected at an early stage of the disease, before the development of symptomatic illness. Heart rate, blood pressure, cardiac output, and systemic vascular resistance showed the greatest changes in the third post-exposure day, correlating with viral load. Applying the classifier model identified participants as flu-positive or negative with an accuracy of 0.81 ± 0.05 two days before major symptoms appeared. Cardiac index and diastolic blood pressure were the leading predicting factors when using data from the first and second day. This study suggests that frequent remote monitoring of advanced physiological parameters may provide early pre-symptomatic detection of flu.


Integrated analysis of pain, health-related quality of life, and analgesic use in patients with metastatic castration-resistant prostate cancer treated with Radium-223.
Radium-223 (Ra-223), an alpha-emitting radiopharmaceutical, established an improved overall survival and health-related quality of life (HRQoL) in symptomatic metastatic castration-resistant prostate cancer (mCRPC) patients. However, effects on pain were not specifically evaluated. Here we assess integrated HRQoL, pain, and opioid use in a contemporary, more extensively pretreated, symptomatic and asymptomatic mCRPC population. mCRPC patients scheduled for Ra-223 treatment were included and analyzed for HRQoL, pain, and opioid use, using Functional Assessment of Cancer Therapy-Prostate (FACT-P) and Brief Pain Inventory-Short Form (BPI-SF) questionnaires and recording of opioid use and dosage, respectively. Primary outcome measure was the percentage of patients experiencing a complete pain response (score of 0 on the BPI-SF Worst pain item and no increase in daily use of analgesics). A complete or partial pain response (better BPI-SF score and decrease in opioid use) and a better or no change in HRQoL was evaluated as an integrated overall clinical response (IOCR). Secondary endpoints included the time to pain progression (TPP) and Total FACT-P deterioration (TTFD), defined as time from first Ra-223 treatment to clinical meaningful increase in BPI-SF Worst pain item score and Total FACT-P score, respectively. This registry included 300 patients, of whom 105 (35%) were evaluable for FACT-P and BPI-SF during Ra-223 treatment. Forty-five (43%) patients had pain at baseline (PAB) (BPI-SF Worst pain score 5-10 points) and 60 (57%) had no pain at baseline (no-PAB) (BPI-SF Worst pain score 0-4 points). Complete pain response was achieved in 31.4% of the patients, while 58% had an IOCR. The median TTP and TTFD were 5.6 and 5.7 months, respectively, while the difference between PAB and no-PAB patients was not significant. In contemporary, extensively pretreated mCRPC patients, Ra-223 treatment induced complete pain responses while integrated analysis of HRQoL, pain response, and opioid use demonstrated that the majority of patients derive clinical benefit.


Delaying, debating and declining motherhood.
Trends of delayed childbearing have accompanied declining birth rates and increasing numbers of childless adults in the USA. Women may postpone parenting in order to save money, find a partner, and get a 'family-friendly' job, but this reproductive strategy may not always be effective. This paper uses two waves of longitudinal data to track childless women's reproductive decision-making and behaviours. During wave 1, interviews were conducted with 72 childless US women between the ages of 25 and 40 about their reproductive desires and intentions. Approximately four years later, a subset of the original sample participated in surveys to assess consistencies between fertility intentions and outcomes, and in-depth interviews to elicit information about changes in their lives that transpired. Whereas some wave 2 participants had fulfilled their goal of becoming parents, the majority were still employing a delaying strategy or had declined to have children. Delayed childbearing was individually strategic for those who could garner resources to be in a better financial or social position to have and raise children, while others kept facing barriers that prevented them from realising their reproductive goals or changed their mind about their fertility intentions and desires.


Alcohol craving in outpatients with alcohol dependence: rate and clinical correlates.
The present study was undertaken to assess the rate and severity of alcohol craving symptoms among registered alcohol-dependent patients at a Veterans Affairs outpatient clinic. We also examined the relationship between alcohol craving and the clinical characteristics of alcohol-dependent patients. Participants included 101 alcohol-dependent veterans enrolled in an outpatient addiction clinic. Alcohol craving was measured by the Penn Alcohol Craving Scale. Alcoholism severity and clinical characteristics were assessed with the Addiction Severity Index, Timeline Followback method, and other instruments. Three alcohol-craving groups (low, moderate, and high) were identified and compared using their demographic and clinical characteristics. Multiple regression analysis was performed to evaluate the relationship between the potential predictors and alcohol craving. The rate of alcohol craving was as follows: low craving (46%), moderate craving (29%), and high craving (25%). When these three craving groups were compared using univariate analyses, patients with higher alcohol craving had significantly higher alcohol composite scores (last 30 days) and severe alcohol dependence (last 1 year). In multiple regression analysis, the model explained 50% of the variance in alcohol craving, with alcoholism severity (42%), withdrawal symptoms (5%), and depression (3%) as significant predictors. Patients in an outpatient treatment setting had a wide range of alcohol cravings. Level of alcohol craving was directly associated with several increased indices of alcohol- and non-alcohol-related morbidity.


Evaluation of the Spider (Phlogiellus genus) Phlotoxin 1 and Synthetic Variants as Antinociceptive Drug Candidates.
Over the two last decades, venom toxins have been explored as alternatives to opioids to treat chronic debilitating pain. At present, approximately 20 potential analgesic toxins, mainly from spider venoms, are known to inhibit with high affinity the NaV1.7 subtype of voltage-gated sodium (NaV) channels, the most promising genetically validated antinociceptive target identified so far. The present study aimed to consolidate the development of phlotoxin 1 (PhlTx1), a 34-amino acid and 3-disulfide bridge peptide of a Phlogiellus genus spider, as an antinociceptive agent by improving its affinity and selectivity for the human (h) NaV1.7 subtype. The synthetic homologue of PhlTx1 was generated and equilibrated between two conformers on reverse-phase liquid chromatography and exhibited potent analgesic effects in a mouse model of NaV1.7-mediated pain. The effects of PhlTx1 and 8 successfully synthetized alanine-substituted variants were studied (by automated whole-cell patch-clamp electrophysiology) on cell lines stably overexpressing hNaV subtypes, as well as two cardiac targets, the hCaV1.2 and hKV11.1 subtypes of voltage-gated calcium (CaV) and potassium (KV) channels, respectively. PhlTx1 and D7A-PhlTx1 were shown to inhibit hNaV1.1-1.3 and 1.5-1.7 subtypes at hundred nanomolar concentrations, while their affinities for hNaV1.4 and 1.8, hCaV1.2 and hKV11.1 subtypes were over micromolar concentrations. Despite similar analgesic effects in the mouse model of NaV1.7-mediated pain and selectivity profiles, the affinity of D7A-PhlTx1 for the NaV1.7 subtype was at least five times higher than that of the wild-type peptide. Computational modelling was performed to deduce the 3D-structure of PhlTx1 and to suggest the amino acids involved in the efficiency of the molecule. In conclusion, the present structure-activity relationship study of PhlTx1 results in a low improved affinity of the molecule for the NaV1.7 subtype, but without any marked change in the molecule selectivity against the other studied ion channel subtypes. Further experiments are therefore necessary before considering the development of PhlTx1 or synthetic variants as antinociceptive drug candidates.


Subependymal Giant Cell Astrocytoma Size Measurement in Tuberous Sclerosis Complex: Noncontrast vs Contrast-Enhanced 3-Dimensional T1-Weighted Magnetic Resonance Imaging (MRI).
Tuberous sclerosis complex is a multisystem genetic disorder characterized by multiorgan hamartomas, seizures, and developmental delay. Current consensus recommendations for tuberous sclerosis complex include brain magnetic resonance imaging (MRI) without and with contrast every 1 to 3 years until age 25 years for subependymal giant cell astrocytoma screening, and more often in patients with known subependymal giant cell astrocytoma. Recent reports in the literature regarding gadolinium deposition have prompted us to investigate the added value of intravenous contrast in this setting. Thirty-nine MRI examinations from 12 tuberous sclerosis complex patients with known subependymal giant cell astrocytoma were retrospectively reviewed for all caudothalamic groove lesions larger than 1 cm. Lesions were measured in 2 planes on both noncontrast and contrast-enhanced axial 3-dimensional (3D) T1-weighted imaging in a randomized fashion by 2 attending pediatric neuroradiologists. Differences in lesion measurements were compared to zero using a 1-sample t test. On average, radiologist A measured lesions 1.1 mm larger in long-axis diameter without contrast (P = .0003), whereas radiologist B measured lesions 0.1 mm larger without contrast (not significant). Differences in lesion measurement were smaller when comparing noncontrast to contrast-enhanced 3D T1-weighted imaging than when comparing radiologist A to radiologist B. Noncontrast axial 3D T1-weighted imaging is similar to contrast-enhanced axial 3D T1-weighted imaging for subependymal giant cell astrocytoma size measurement. It has high accuracy and may be a safer and more efficient imaging method for tuberous sclerosis complex patients who commonly undergo numerous MRI examinations throughout their lifetime. Further studies may be appropriate to determine whether contrast material could be beneficial in certain situations.


Welfare in horse breeding.
Welfare problems related to the way horses are bred, whether by coitus or by the application of artificial reproduction techniques (ARTs), have been given no discrete consideration within the academic literature. This paper reviews the existing knowledge base about welfare issues in horse breeding and identifies areas in which data is lacking. We suggest that all methods of horse breeding are associated with potential welfare problems, but also that the judicious use of ARTs can sometimes help to address those problems. We discuss how negative welfare effects could be identified and limited and how positive welfare effects associated with breeding might be maximised. Further studies are needed to establish an evidence base about how stressful or painful various breeding procedures are for the animals involved, and what the lifetime welfare implications of ARTs are for future animal generations.


Differential expression patterns of Wnt and beta-catenin/TCF target genes in the uterus of immature female rats exposed to 17alpha-ethynyl estradiol.
To characterize the effects of an estrogen receptor (ER) agonist on the gene expressions in the uterus, immature female rats were administered once orally with 17alpha-ethynyl estradiol (EE, 3 mug/kg), a potent ER agonist. We focused on four categories of sex steroid hormone receptor genes: well-known estrogen target genes, Wnt genes, and beta-catenin/T-cell factor (TCF) target genes. ERalpha, ERbeta, progesterone receptor, and androgen receptor mRNAs were all downregulated at 24 and/or 48 h after EE administration. Complement C3 and insulin-like growth factor 1 mRNAs were markedly induced after EE administration. Although the time courses of Wnt4, Wnt5a, and Wnt7a mRNA status varied until 12 h after EE administration, all of them were simultaneously downregulated at 24 and 48 h. The remarkable downregulation of Wnt7a mRNA in response to EE was considered to be important to understand the various uterine phenomena affected by ER agonists. In the beta-catenin/TCF target genes, the downregulation of anti-Mullerian hormone type 2 receptor and bone morphogenetic protein 4 mRNA after EE administration appeared to be closely related to the downregulation of Wnt7a. The upregulation of cyclin D1 and follistatin mRNA at the early phase after EE administration was considered to have been affected by the upregulation of Wnt4. These results indicate that an ER agonist influences not only the mRNA expression of sex steroid hormone receptor genes and well-known estrogen target genes but also Wnt genes (Wnt4, Wnt5a, Wnt7a) and beta-catenin/TCF target genes in the uterus of immature rats, indicating that their molecules are the potential players affected by estrogenic stimuli.