Clathrin adaptor AP-1-mediated Golgi export of amyloid precursor protein is crucial for the production of neurotoxic amyloid fragments.

One of the hallmarks of Alzheimer's disease is the accumulation of toxic amyloid-ß (Aß) peptides in extracellular plaques. The direct precursor of Aß is the carboxyl-terminal fragment ß (or C99) of the amyloid precursor protein (APP). C99 is detected at elevated levels in Alzheimer's disease brains, and its intracellular accumulation has been linked to early neurotoxicity independently of Aß. Despite this, the causes of increased C99 levels are poorly understood. Here, we demonstrate that APP interacts with the clathrin vesicle adaptor AP-1 (adaptor protein 1), and we map the interaction sites on both proteins. Using quantitative kinetic trafficking assays, established cell lines and primary neurons, we also show that this interaction is required for the transport of APP from the trans-Golgi network to endosomes. In addition, disrupting AP-1-mediated transport of APP alters APP processing and degradation, ultimately leading to increased C99 production and Aß release. Our results indicate that AP-1 regulates the subcellular distribution of APP, altering its processing into neurotoxic fragments.

Repurposing of 5-nitrofuran-3,5-disubstituted isoxazoles: A thriving scaffold to antitrypanosomal agents.

Chagas disease (CD) is a neglected disease caused by the protozoan Trypanosoma cruzi. The two drugs used in the treatment schedules exhibit adverse effects and severe toxicity. Thus, searching for new antitrypanosomal agents is urgent to provide improved treatments to those affected by this disease. 5-Nitrofuran-isoxazole analogs were synthesized by cycloaddition reactions [3+2] between chloro-oximes and acetylenes in satisfactory yields. We analyzed the structure-activity relationship of the analogs based on Hammett's and Hansch's parameters. The 5-nitrofuran-isoxazole analogs exhibited relevant in vitro antitrypanosomal activity against the amastigote forms of T. cruzi. Analog 7s was the trending hit of the series, showing an IC50 value of 40 nM and a selectivity index of 132.50. A possible explanation for this result may be the presence of an electrophile near the isoxazole core. Moreover, the most active analogs proved to act as an in vitro substrate of type I nitroreductase rather than the cruzain, enzymes commonly investigated in molecular target studies of CD drug discovery. These findings suggest that 5-nitrofuran-isoxazole analogs are promising in the studies of agents for CD treatment.

[Detecting of obstructive sleep apnea hypopnea syndrome using a multi-parameter pressure sensitive sleep monitor].

OBJECTIVE: To evaluate the efficacy of an electro-mechanical film-based(EMFi) multi-parameter pressure sensitive sleep monitor(MPSSM)on clinical diagnosis and research significance of obstructive sleep apnea hypopnea syndrome(OSAHS). METHODS: Retrospective analysis was made of 58 test subjects at Peking University Third Hospital with suspected OSAHS who were simultaneously monitored by MPSSM and polysomnography(PSG). The PSG test results were used as the gold standard in evaluating the sensitivity and specificity of OSAHS diagnosis of MPSSM. The test result consistency of sleep apnea and hypopnea index(AHI)and total apnea time of the two methods was evaluated. Real-time waveform comparison of sleep respiratory events of a randomly selected patient diagnosed with OSAHS was performed. RESULTS: For 58 test subjects, 48 were male, 10 were female, with an average age of(40.6±12.2)years. Thirty-nine out of the 58 test subjects were diagnosed with OSHAS by PSG. The sensitivity of MPSSM for OSAHS diagnosis was 92.3%, with 95% confidence interval of 79.1%-98.4%, and the specificity of MPSSM for OSAHS diagnosis was 100%, with 95% confidence interval of 82.3%-100%. Kappa test k=0.887 (P < 0.001) showed OSAHS diagnosis results of the two methods were almost identical. The AHI measured by MPSSM [12.0(2.6-32.2) times/h] and PSG [13.4(3.1-38.8) times/h] were highly correlated (ρ=0.939, P < 0.001). The total apnea time measured by MPSSM [37.9(9.9-80.5) min] and PSG [32.3(8.6-93.0) min] were highly correlated(ρ=0.924, P < 0.001). Bland-Altman plot showed that the consistency between the test results of the two methods was very high. CONCLUSION: As a portable, non-contact, fully automatic monitoring device, MPSSM is reliable in the screening of OSAHS compared with PSG. It is suitable to be promoted and applied in primary medical institutions, nursing homes and domestic usage. However, further research is required in improving the analysis of different sleep phase and the differentiation of central sleep apnea syndrome respiratory events in order to effectively assist medical personnel in making an accurate sleep apnea diagnosis.

Timing carbon turnover (δ13C) in weaned piglet's brain by IRMS.

Isotope-ratio mass spectrometry (IRMS) is a potential tool that provides time-integrated estimate of assimilated and not just ingested nutrients. This feature turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ13C) in the brain of weanling piglets. Eighty-seven piglets weaned at 21-day-old were used, being three piglets slaughtered at day-0 (prior to diet-switch) and, the remaining 84 randomly assigned in a complete block design with a 2 x 2 factorial arrangement of treatments (two nucleotide levels: 0% and 0.1% and, two glutamate levels: 0% and 1%). On trial days 3, 6, 9, 14, 21, 35 and 49, three piglets per treatment were also slaughtered. The samples were analyzed by IRMS and adjusted to the first-order equation by a nonlinear regression analysis using NLIN procedure of SAS, in order to obtain the exponential graphics. The carbon turnover (t95%) verified for cerebral tissue was faster (P<0.05) for diet containing glutamate in comparison to other diets, supporting the fact that glutamate contributed to develop the piglets' brain, due to the fastest incorporation of 13C-atoms in this tissue at post-weaning phase, despite the energy deficit experienced by them.

Atmospheric ammonia alters lipid metabolism-related genes in the livers of broilers (Gallus gallus).

Atmospheric ammonia in animal housing is reported to have adverse effects on livestock performance and animal health. Previous experiments have found that 75 ppm ammonia reduced the production performance and altered body fat distribution quality of broilers. In this study, we examined the body fat distribution, serum metabolites and lipid metabolism gene expression of broiler exposed to ammonia. A total of 400 chickens were randomly allocated to four groups with four replicates and received ammonia treatments at 0, 25, 50 and 75 ppm, respectively, for 3 weeks. The average daily feed intake and weight gain were decreased when broiler was exposed to ammonia concentration exceeding 50 ppm (p < .05). The increased abdominal fat and reduced thickness of subcutaneous adipose were found in broilers of 75 ppm group (p < .05). When ammonia exceeded 50 ppm, the content of fat in breast muscle of broiler was increased, and when ammonia was higher than 25 ppm, the fat in liver was increased (p < .05). It showed that the fat content in liver was a sensitive index for broilers exposed to ammonia. Furthermore, ammonia exposure had no significant effect on total cholesterol and triglyceride in serum, but significantly increased the relative mRNA expression of acetyl-CoA carboxylase (p = .046) and malic enzyme in liver (p = .038), which indicated that ammonia exposure may increase the de novo fat synthesis in liver. In addition, ammonia increased the high-density lipoprotein cholesterol (p = .02) and activity of hepatic lipase in serum (p < .001), which indicated that ammonia exposure may improve the transportation of cholesterol to liver. To conclude, our results indicated that ammonia exposure might increase the de novo fat synthesis in liver and increased the transportation of cholesterol to liver. In addition, the concentration of ammonia in poultry house should be limited lower than 25 ppm based on the variation of hepatic fat content.

Changes of expression of estrogen and progestrone receptors, human epithelial growth factor receptor 2 and Ki-67 after neoadjuvant chemotherapy in the treatment of breast cancer.

Recent studies suggest that the development and prognosis of breast cancer is in close correlation to molecular subtype of breast cancer. Neoadjuvant chemotherapy has been extensively applied in the treatment of local breast cancer in advanced stage. In order to verify the correlation between expression changes of estrogen receptor, progestrone receptor, human epithelial growth factor receptor 2 and Ki-67 after neoadjuvant chemotherapy and neoadjuvant chemotherapy, we studied 120 patients with stage IIAIIIC breast cancer who underwent neoadjuvant chemotherapy in Binzhou Medical University Hospital, Shandong, China from February 2011 to February 2015. Clinical characteristics were retrospectively analyzed. The expression of estrogen receptor, progesterone receptor, human epithelial growth factor receptor 2 and Ki-67 of patients were detected using the immunohistochemical method before and after neoadjuvant chemotherapy. The results suggest that the overall remission rate of neoadjuvant chemotherapy was 76.7% (92/120) of which 16.7% (20/120) of cases had complete remission, 60% (72/120) had partial remission and 23.3% (28/120) were stable. There were no cases of progressive disease. The property of estrogen receptor and the expression of Ki-67 of primary tumor were correlated to the remission rate of neoadjuvant chemotherapy (P less than 0.05). The expression of Ki-67 had a significant decline after neoadjuvant chemotherapy, and the difference had statistical significance (P less than 0.05). The difference in expression of estrogen receptor, progesterone receptor and human epithelial growth factor receptor 2 before and after neoadjuvant chemotherapy had statistical significance (P > 0.05). Hence, it can be concluded that breast cancer patients with negative estrogen receptor expression and high Ki-67 expression before neoadjuvant chemotherapy can achieve better curative effects. Neoadjuvant chemotherapy cannot change the expression states of estrogen receptor, progesterone receptor and human epithelial growth factor receptor 2, but it can lower the expression level of Ki-67. Ki-67 can also be used for predicting the curative effect of neoadjuvant chemotherapy.

Effect of a grain challenge on ruminal, urine, and fecal pH, apparent total-tract starch digestibility, and milk composition of Holstein and Jersey cows.

The effects of a grain challenge on ruminal, urine, and fecal pH, apparent total-tract starch digestibility, and milk composition were determined. Six Holstein cows, 6 rumen-cannulated Holstein cows, and 6 Jersey cows were used in a replicated 3 × 3 Latin square design balanced to measure carryover effects. Periods (10 d) were divided into 4 stages (S): S1, d 1 to 3, served as baseline with regular total mixed ration ad libitum; S2, d 4, served as restricted feeding, with cows offered 50% of the total mixed ration fed on S1 (dry matter basis); S3, d 5, a grain challenge was performed, in which cows were fed total mixed ration ad libitum and not fed (CON) or fed an addition of 10% (MG) or 20% (HG) pellet wheat-barley (1:1) top-dressed onto the total mixed ration, based on dry matter intake obtained in S1; S4, d 6 to 10, served as recovery stage with regular total mixed ration fed ad libitum. Overall, cows had a quadratic treatment effect for milk yield where CON (22.6 kg/d) and HG (23.5 kg/d) had lower milk yield than cows in MG (23.7 kg/d). Jersey cows had a quadratic treatment effect for dry matter intake where cows in CON (13.2 kg/d) and HG (12.4 kg/d) had lower dry matter intake than cows in MG (14 kg/d). Holstein cows had a linear treatment effect for dry matter intake (17.7, 18.4, and 18.6 kg/d for CON, MG, and HG, respectively). Rumen pH for the rumen-cannulated cows had a linear treatment effect (6.45, 6.35, and 6.24 for CON, MG, and HG, respectively). Cows in HG spent more time with rumen pH below 5.8 (4.33 h) than MG (2 h) or CON (2.17 h) as shown by the quadratic treatment effect. Holstein cows in HG (8.46) had lower urine pH than MG (8.51) or CON (8.54) as showed by the linear treatment effect for urine pH. Apparent total-tract starch digestibility had a tendency for a linear treatment effect on S3 (97.62 ± 1.5, 97.47 ± 1.5, and 91.84 ± 1.6%, for CON, MG, and HG, respectively). Fecal pH was associated with rumen pH depression as early as 15 h after feeding for Holstein cows. In conclusion, a grain challenge reduced urine pH in Holstein cows but not in Jersey cows. Holstein cows' health were not affected when rumen pH was depressed. A potentially useful link between rumen pH and systemic (urine) pH within 2 h after feeding was quantified in Holstein cows.

Popular medicinal uses of Calea uniflora Less. (Asteraceae) and its contribution to the study of Brazilian medicinal plants.

Calea uniflora Less. is widely used in southern Santa Catarina (Brazil), but there are no scientific studies which support its use. Then, this study was proposed to determine of the percentage use of C. uniflora in a city of southern Brazil and documentation of the knowledge that the population has about this species. The survey was conducted with semi-structured interviews using a questionnaire applied to 372 participants. In analyzing the results, it was observed that of the 94.1% who recognized C. uniflora, 74.3% utilize it as a medicinal plant and 65.4% of such knowledge originates in childhood, mainly through the family (84.6%). 93% reported using inflorescences macerated in alcohol or rum; this extract is generally used topically for wound healing and muscle pain. Furthermore, some reported using small quantities of this extract orally to treat cold and flu. Regarding effectiveness and safety, 97% stated an improvement in symptoms with the use of the plant, while 98.5% stated that it has no toxicity. In light of these results, future phytochemical, pharmacological and toxicological analyses should be designed in order to ensure rational and safe use of this species.

Effects of direct-fed Bacillus pumilus 8G-134 on feed intake, milk yield, milk composition, feed conversion, and health condition of pre- and postpartum Holstein cows.

The usage of direct-fed microbials (DFM) has become common in the dairy industry, but questions regarding choice of strain, mode of action, and efficacy remain prevalent. The objective of this study was to evaluate the effects of a DFM (Bacillus pumilus 8G-134) on pre- and postpartum performance and incidence of subclinical ketosis in early lactation. Forty-three multiparous Holstein cows were assigned to 2 treatments in a randomized complete block design; cows in the direct-fed microbial treatment (DFMt, n=21) received 5.0×10(9) cfu/cow of B. pumilus in 28 g of a maltodextrin carrier, whereas cows in the control treatment (CON, n=22) received 28 g of maltodextrin carrier alone. Treatments were top-dressed on the total mixed ration daily. Treatments were applied from 21 d before expected calving date to 154 d after calving. Cows on treatment DFMt tended to have lower serum haptoglobin concentration than CON cows on d 14. Cows on treatment DFMt had higher IgA concentrations in milk than CON cows during the first week after calving. Cows fed DFMt had higher yields of milk, fat-corrected milk, energy-corrected milk, milk fat, and milk protein during the second week of lactation than CON; however, we found no differences between treatments on milk yield and milk components overall. Cows on DFMt tended to have higher feed conversion and to have lower prevalence of subclinical ketosis (beta-hydroxybutyrate >1.2 mmol/L) on d 5 than cows fed CON. Dry matter intake, body weight, and body condition score were not affected by DFMt supplementation. Milk production efficiencies (calculated based on fat-corrected milk and energy-corrected milk) were higher by 0.1 kg of milk per kilogram of dry matter intake in cows that received DFMt compared with cows that received CON. In conclusion, cows receiving DFMt tended to have lower incidence of subclinical ketosis than cows receiving CON. Cows fed DFMt tended to have higher feed conversion and evidence for greater immunity than CON. Supplementation with B pumilus 8G-134 may provide benefits for transition cow health and milk production efficiency.

The PGI-KLF4 pathway regulates self-renewal of glioma stem cells residing in the mesenchymal niches in human gliomas.

Gliomas display cellular hierarchies with self-renewing tumorigenic glioma stem cells (GSCs) at the apex. The GSC niches function as a regulator of GSC maintenance, however, the exact components of GSC niches that mediate this process are still far from fully defined. Here, we showed that glioma cells with aberrant mesenchymal phenotypes constitute a mesenchymal niche for GSCs. Using patient-derived specimens, we demonstrated that the paracrine PGI signaling, initiated by mesenchymal glioma cells, induces the self-renewal and tumorigenic potentials of GSCs through induction of KLF4. Treatment of intracranial orthotopic xenografts with shPGI or shKLF4 leads to less lethal potency. Our data therefore suggest that blockade of the PGI-KLF4 pathway may provide a therapeutic strategy against GSC niches.

Poly(methylmethacrylate-co-dimethyl acrylamide)-silver nanocomposite prevents biofilm formation in medical devices.

Aim: To investigate whether medical devices coated with a synthesized nanocomposite of poly(methylmethacrylate-co-dimethyl acrylamide) (PMMDMA) and silver nanoparticles (AgNPs) could improve their antibiofilm and antimicrobial activities. We also investigated the nanocomposite's safety. Materials & methods: The nanocomposite was synthesized and characterized using analytical techniques. Medical devices coated with the nanocomposite were evaluated for bacterial adhesion and hemolytic activity in vitro. Results: The nanocomposite formation was demonstrated with the incorporation of AgNPs into the polymer matrix. The nanocomposite proved to be nonhemolytic and significantly inhibited bacterial biofilm formation. Conclusion: The PMMDMA-AgNPs nanocomposite was more effective in preventing biofilm formation than PMMDMA alone and is a promising strategy for coating medical devices and reducing mortality due to hospital-acquired infections.

[Box: see text].

[Extent of lymphadenectomy for esophageal cancer].

Esophageal cancer is a common malignant tumor in China. For resectable ones, surgery is still the primary treatment. At present, the extent of lymph node dissection remains controversial. Extended lymphadenectomy makes metastatic lymph nodes more likely to be resected, which contributed to pathological staging and postoperative treatment. However,it may also increase the risk of postoperative complications and affect prognosis. Therefore, it is controversial how to balance the optimal extent/number of dissected lymph nodes for radical resection with the lower risk of severe complications. In addition, whether the lymph node dissection strategy should be modified after neoadjuvant therapy needs to be investigated, especially for patients who have a complete response to neoadjuvant therapy. Herein, we summarize the clinical experience on the extent of lymph node dissection in China and worldwide, aiming to provide guidence for the extent of lymph node dissection in esophageal cancer.

Bacillus amyloliquefaciens spray improves the growth performance, immune status, and respiratory mucosal barrier in broiler chickens.

Studies have shown that supplementing probiotics in a nasal spray can provide prophylaxis against upper respiratory tract infections. Previously, we found that spraying with Bacillus reduced the concentration of atmospheric ammonia and airborne microbiological aerosols in broiler houses. In the present study, we evaluated the effects of spraying with Bacillus amyloliquefaciens on the respiratory mucosal barrier in broilers. Two trails were conducted simultaneously in four environmentally controlled chambers. In each trail, 320 1-d-old male broilers were randomly assigned to control (CON) and B. amyloliquefaciens (BAQ) group. Eight cages containing 20 birds per cage were placed in two chambers. The BAQ group chambers were sprayed with B. amyloliquefaciens at 4 × 109 CFU/m2 each day for 42 d. Spraying with B. amyloliquefaciens increased average daily gain and average daily feed intake in broilers (P < 0.01), but had no effects on FCR (P > 0.05). The relative weights of the bursa (21 d) and spleen (42 d) were higher in the BAQ group (P < 0.05). The serum IgG, IgM (21 d), and IgA, Ig M (42 d) levels were improved in BAQ group (P < 0.05). The sIgA levels in tracheal lavage fluid were significantly higher in the BAQ group than the CON group (P < 0.05). Furthermore, treatment with BAQ downregulated IL-6 and IFN-γ (P < 0.05), but upregulated IL-2, occludin, and mucin-2 (P < 0.05). In addition, the number of acidic goblet cells was significantly higher in the tracheas of treated broilers than untreated broilers (P < 0.05). In conclusion, spraying with B. amyloliquefaciens may help to enhance the growth performance and improve the respiratory mucosal barrier in broiler chickens.

Voltage-dependent K+ channels as targets of osmosensing in guard cells

Guard cell turgor responds to the osmogradient across the plasma membrane and controls the stomatal aperture. Here, we report that guard cells utilize voltage-dependent K+ channels as targets of the osmosensing pathway, providing a positive feedback mechanism for stomatal regulation. When exposed to a hypotonic condition, the inward K+ current (IKin) was highly activated, whereas the outward K+ current (IKout) was inactivated. In contrast, hypertonic conditions inactivated the IKin while activating IKout. Single-channel recording analyses indicated that an alteration in channel opening frequency was responsible for regulating IKin and IKout under different osmotic conditions. Further studies correlate osmoregulation of IKin with the pattern of organization of actin filaments, which may be a critical component in the osmosensing pathway in plant cells.

The glucocorticoid inhibits neutrophils formed extracellular traps (NETs) and suppresses the inflammation caused by fallopian tube staphylococcal infection.

OBJECTIVE: Fallopian tube can transport zygote into the uterine cavity, while inflammation may cause certain influences on the fallopian tube's function. Neutrophils formed extracellular traps (NET) can kill pathogenic microorganisms. This study intends to analyze the role of glucocorticoid in the regulation of NETs sterilization during the fallopian tube staphylococcal infection. MATERIALS AND METHODS: Rat fallopian tube staphylococcal infection model was established. Group A was the control group, group B was the model group, and group C was the dexamethasone intervention group. ELISA was applied to test inflammatory factors, including citrullinated histone H3 (CitH3) and high molecular weight kininogen (HK) content in serum. RT-PCR was performed to test the mRNA expression of glucocorticoid receptor α, ß (GR-α, GR-ß). Western blot was used to detect the protein levels of GR-α and GR-ß. RESULTS: Microscopically, group A showed clear fallopian tube wall and unobstructed lumen. Group B presented obscured tube wall, blocked lumen, and inflammatory cells infiltration. Group C demonstrated unclear tube wall and a few inflammatory cells infiltration. Serum CitH3 level was increased, while HK was down-regulated in group B compared with group A. CitH3 was declined, whereas HK was enhanced in group C compared with group B (p<0.05). The mRNA expression of GR-ß was reduced, while GR-α expression was elevated in group C compared with group A and B. Group B showed upregulated GR-ß expression and reduced GR-α mRNA and protein expression compared with group A (p<0.05). CONCLUSIONS: Rat fallopian tube Staphylococcus aureus infection activates NETs, elevates CitH3, and decreases HK. Glucocorticoid can inhibit inflammation through down-regulate GR-ß and up-regulate GR-α expression.

Publisher's Note: Laser propagation in dense magnetized plasma [Phys. Rev. E 94, 053207 (2016)].

This corrects the article DOI: 10.1103/PhysRevE.94.053207.

Laser propagation in dense magnetized plasma.

A right-hand circularly polarized electromagnetic wave can propagate in a sufficiently magnetized plasma of any density without encountering cutoff in the whistler mode. With the recent realization of tens-kilotesla magnetic fields, laser propagation in highly magnetized high-density plasmas has become of practical interest, especially for heating plasmas to high energy density and igniting fusion targets. In this paper, the whistler regime of laser-plasma interaction is discussed. It is shown by one- and two-dimensional particle-in-cell simulations that moderately intense right-hand circularly polarized laser light can enter and propagate in high-density plasma and heat it efficiently because of the significantly reduced wave length and speed.

PACS-2 mediates the ATM and NF-κB-dependent induction of anti-apoptotic Bcl-xL in response to DNA damage.

Nuclear factor kappa B (NF-κB) promotes cell survival in response to genotoxic stress by inducing the expression of anti-apoptotic proteins including Bcl-xL, which protects mitochondria from stress-induced mitochondrial outer membrane permeabilization (MOMP). Here we show that the multifunctional sorting protein Pacs-2 (phosphofurin acidic cluster sorting protein-2) is required for Bcl-xL induction following DNA damage in primary mouse thymocytes. Consequently, in response to DNA damage, Pacs-2(-/-) thymocytes exhibit a blunted induction of Bcl-xL, increased MOMP and accelerated apoptosis. Biochemical studies show that cytoplasmic PACS-2 promotes this DNA damage-induced anti-apoptotic pathway by interacting with ataxia telangiectasia mutated (ATM) to drive NF-κB activation and induction of Bcl-xL. However, Pacs-2 was not required for tumor necrosis factor-α-induced NF-κB activation, suggesting a role for PACS-2 selectively in NF-κB activation in response to DNA damage. These findings identify PACS-2 as an in vivo mediator of the ATM and NF-κB-dependent induction of Bcl-xL that promotes cell survival in response to DNA damage.