Search

The recapitulation of complex microenvironments that regulate cell behavior during development, disease, and wound healing is key to understanding fundamental biological processes. In vitro, multicellular morphogenesis, organoid maturation, and disease modeling have traditionally been studied using either non-physiological 2D substrates or 3D biological matrices, neither of which replicate the spatiotemporal biochemical and biophysical complexity of biology. Here, we provide a guided overview of the recent advances in the programming of synthetic hydrogels that offer precise control over the spatiotemporal properties within cellular microenvironments, such as advances in the control of cell-driven remodeling, bioprinting, or user-defined manipulation of properties (e.g., via light irradiation).

ABSTRACTImbalance in production and clearance of amyloid beta (Aβ) is the primary reason for its deposition in Alzheimer disease. Macroautophagy/autophagy is one of the important mechanisms for clearance of both intracellular and extracellular Aβ. Here, through screening, we identified alborixin, an ionophore, as a potent inducer of autophagy. We found that autophagy induced by alborixin substantially cleared Aβ in microglia and primary neuronal cells. Induction of autophagy was accompanied by up regulation of autophagy proteins BECN1/Beclin 1, ATG5, ATG7 and increased lysosomal activities. Autophagy induced by alborixin was associated with inhibition of the phosphoinositide 3-kinase (PI3K)-AKT pathway. A knock down of PTENand consistent, constitutive activation of AKTinhibited alborixin-induced autophagy and consequent clearance of Aβ. Furthermore, clearance of Aβ by alborixin led to significant reduction of Aβ-mediated cytotoxicity in primary neurons and differentiated N2a cells. Thus, our findings put forward alborixin as a potential anti-Alzheimer therapeutic lead.AbbreviationsAβ: amyloid beta; ALB: alborixin; ATG: autophagy-related; BECN1: beclin 1; DAPI: 4, 6-diamidino-2-phenylindole; DCFH-DA: 2,7-dichlorodihydrofluorescein diacetate; fAβ: fibrillary form of amyloid beta; GFAP: glial fibrillary acidic protein; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MAP2: microtubule-associated protein 2; MTOR: mechanistic target of rapamycin kinase; PTEN: phosphatase and tensin homolog; ROS: reactive oxygen species; SQSTM1: sequestosome 1; TMRE: tetramethylrhodamine, ethyl ester

The article discusses civil-military relations in Pakistan after dismissal of Nawaz Sharif as Prime Minister of Pakistan. Topics include diplomatic and unbalanced relations between armed forces and civilian government; attempt made by People's Party for placing Inter-Services Intelligence under Interior ministry; and attempts made by Indian prime minister Narendra Modi for isolating Pakistan.

AbstractSalvia officinalisis a versatile medicinal plant with multitude of applications. The present study was designed to study the variation in the quality and quantity of the leaf essential oil of S. officinaliscollected during all the months between July 2012 and June 2013, along with its antioxidant and antibacterial activity. The highest percentage of essential oil was found in the month of July (0.600%) and the lowest percentage of oil was recorded in the month of September (0.225%), a period of latent vegetation. The percentage of major constituents of twelve sage samples were: α-pinene (3.22-7.69%), β-pinene (1.57-28.33%), camphene (1.22-6.11%), α-thujone (17.20-40.45%), β-thujone (2.17-4.97%), 1,8-cineole (0.97-7.76%), camphor (0.88-36.97%), borneol (2.12-18.38%), α-caryophyllene (1.38-17.32%) and β-caryophyllene (1.15-15.37 %). The relative percentage of camphor was higher (0.88-36.97%) during cold winters (Nov-Feb). Borneol, αand β-caryophyllene were also at higher percentage in the vegetative period (May-June) while as α-thujone and β-thujone were at peak at the end of vegetative period. The essential oil samples exhibited broad spectrum antibacterial activity against the gram positive bacterial pathogens and showed moderate free radical scavenging activity.

Previous investigations provide evidence of an association of hypogonadism with type 2 diabetes in men, and low testosterone levels have been regarded a risk factor for the disease. Since a strong genetic predisposition to type 2 diabetes has been demonstrated, here we investigate a possible tendency towards hypogonadism in young male offspring of diabetic parents.

The article focuses on the conflict between India and Pakistan over territorial claims in Kashmir. It mentions the visitation of Indian Prime Minister Narendra Modi to Kashmir which has become an integral part of India. It notes that Pakistan maintains Kashmiris' right to self-determination through a plebiscite and the promised plebiscite should be allowed to decide the fate of the Kashmiri people.

Four patients from three families with the clinical features of DOOR syndrome (onycho-osteodystrophy, dystrophic thumbs, sensorineural deafness, and increased urinary levels of 2-oxoglutarate) are the subjects of this report. Our report deals with the autosomal recessive form of the disease, wherein the activity of 2-oxoglutarate decarboxylase (E10) in fibroblasts and white blood cells of the patients is decreased. The activity of E10 in all patients' fibroblasts and white blood cells was significantly lower compared to the controls. This study demonstrates for the first time that E10 deficiency is an important biochemical marker for the autosomal recessive form of DOOR syndrome. © 2002 Wiley-Liss, Inc.

Objectives:Develop a natural language processing (NLP) system for identification of patients with peripheral artery disease (PAD).Background:Despite its high prevalence and clinical impact, research on PAD remains limited due to poor accuracy of billing codes. Ankle and toe-brachial index (ABI, TBI) can be used to identify PAD patients with high accuracy using electronic health record (EHR) data.Methods:A random sample of 800 ABI test reports from 94 Veterans Affairs (VA) facilities during 2015-2017 were selected and annotated by clinical experts. We trained the NLP system using random forest models and optimized it through sequential iterations of 10-fold cross validation and error-analysis on 600 test reports and evaluated its final performance on a separate set of 200 reports. We also assessed the accuracy of NLP-extracted ABI and TBI values for identifying patients with PAD in a separate cohort undergoing ABI testing.Results:The NLP system had an overall precision (positive predictive value) of 0.85, recall (sensitivity) of 0.93 and F1-measure (overall accuracy) of 0.89 to correctly identify ABI/TBI values and laterality. Among 261 patients with ABI testing (49% PAD), the NLP system achieved a positive predictive value of 92.3%, sensitivity of 83.1% and specificity of 93.1% to identify PAD when compared to a structured chart review (Table). The above findings were consistent in a range of sensitivity analysis (Table).Conclusion:We successfully developed and validated an NLP system for identifying patients with PAD within the VA’s EHR. Our findings have broad implications for PAD research and quality improvement efforts.

This paper describes the development of a new method for monitoring acetic anhydride in the presence of acetic acid in the occupational environment. The method consisted of collecting airborne acetic anhydride on a solid sorbent, desorbing it with acetone and analyzing it by gas chromatography. During the development of this method various parameters such as selection of column, selection of solid sorbent, selection of desorber solvent, effects of humidity, flow rate, total sample volume, desorption efficiency, and shelf-life of samples were studied. Interference from methyl acetate, water, formic acid, and acetic acid was also tested with the determination of acetic anhydride.

A novel method for monitoring occupational exposure to ethylene oxide was developed. Ethylene oxide was collected on Columbia JXC activated carbon, desorbed with carbon disulfide, and analyzed by gas chromatography using a TERGITOL TMN or UCON LB550X column. Several parameters such as selection of adsorbent and column; effects of humidity, sampling rate and volume; shelf-life; interference from ten contaminants; desorption technique and calibration curve were investigated. The lower detection limit of the method is 0.15 ppm in ten liters of air. The method was evaluated statistically and by conducting a round-robin interlaboratory study.

Metabolic labelling of Plasmodium falciparum parasites with [3H]GIcN, [3H]Man, [3H]Gal and [3H]ethanolamine, and subsequent purification by SDS‐PAGE of the labelled material provided effective labelling of the MSP‐1, 195 kDa, and MSP‐2, 42‐53 kDa, glycoproteins. Reductive β‐elimination of the MSP‐2 released from the gel consisted of glycopeptides containing labelled sugars. Processing of the eliminated components and identification of the sugar residues demonstrated the presence of N‐acetylglucosaminitol and N‐acetylgalactosaminitol amongst other labelled sugars. Reductive β‐elimination with sodium hydroxide‐sodium borotritide‐borohydride showed the presence of glucosaminitol and alanine in the hydrolysis products. The MSP‐2 was retained on solid phase wheat‐germ agglutinin and was released from the lectin by treatment with GIcNAc. Upon treatment with O‐glycanase the MSP‐2 glycoprotein released lablled amino sugar, and derived oligosaccharides on treatment with exoglycosidases released labelled components corresponding to the metabolically incorporated sugars. Labelled Gal was incorporated into the MSP‐2 glycoprotein using [3H]UDP‐Gal and galactosyltransferase. The galactosylated glycoprotein released labelled Gal upon treatment with β‐galactosidase. The results of the present study suggest that the carbohydrate chains of the MSP‐2 glycoprotein are attached to the protein backbone via GlcNAc‐ and GalNAc‐sedne/threonine in O‐glycosyl linkage and the glycoprotein has terminal GlcNAc and Gal residues. The carbohydrate moieties of MSP‐2, glycoprotein consist mainly of short chains linked to the protein core.

Extraction of vanadium(V) from acidic sulfate solutions by a mixture of tri-n-octylamine and tributylphosphate (used as modifier) dissolved in kerosene has been studied. The distribution coefficient of vanadium(V) increases with an increase in pH and vanadium(V) concentration. The presence of iron(III) in aqueous solution does not have any appreciable effect, while large amounts of sulfate ion depress the distribution coefficient. Uranium(VI) has a distribution different from vanadium(V). Based upon these results, a scheme for the separation of vanadium(V) from uranium leach liquors has been made and checked experimentally.

Five highly purified human pituitary LH preparations (candidate preparations for a new International Reference Preparation (IRP) and coded A to E) and three highly purified commercial preparations (referred to as Kabi I, II and III) were fractionated by electrofocusing on a sucrose density gradient with ampholytes in the pH range of 3·5–10·0 The LH activity was monitored in each of the eluted fractions by an in-vitro bioassay and a radioimmunoassay procedure and the profiles of biological activity and immunoreactivity were compared with those of the first IRP of Human Pituitary Luteinizing Hormone for Immunoassay (code no. 68/40).The overall recovery of the biological activity after electrofocusing of the nine preparations ranged between 75 and 92% (mean 86%) and was higher (P< 0·05) than that of the immunological reactivity which varied between 71 and 94% (mean 79%).The profiles of the biological activity and immunological reactivity were in close agreement with each other in all preparations. Significant differences were, however, observed in the distribution of various molecular species between the currently used standard and the eight other highly purified preparations.The ratios of biological activity (B) to immunoreactivity (I) of preparation B and of the three Kabi preparations were significantly higher than those of the other five preparations, both before and after electrofocusing. After electrofocusing, a slight but significant (P< 0·05) rise in the B/I ratios was observed, suggesting that some biologically inactive immunoreactive material was removed during fractionation. All purified preparations lacked the characteristic 'acidic' human LH species which accompanies FSH and which is abundant in the IRP (69/104) and is also present in aqueous extracts of postmenopausal pituitary glands.A comparison of the electrofocusing profiles of the nine highly purified preparations with those of human LH from plasma and individual pituitary glands revealed marked differences, suggesting that the methods used for the purification of the hormone significantly altered its molecular composition.

The androgen dependence and regional distribution of specific epididymal proteins have been investigated in the adult rhesus monkey (Macaca mulatta). The protein pattern of the epididymal cytosol was studied in intact, castrated, and testosterone-treated castrated monkeys by polyacrylamide gel electrophoresis (PAGE) and electrofocusing. At least five distinct protein components of the epididymal cytosol were androgen-sensitive. The PAGE of cytosol from various portions of the epididymis showed that whereas four protein components were uniformly distributed in the caput, corpus, and caudal segments, one of the androgen-dependent proteins was found to be associated with the caudal portion of the epididymis. Electrofocusing of epididymal cytosol also confirmed the presence of five androgen-dependent proteins in the adult monkey. The isoelectric points of these proteins were shown to range between 5.6 and 6.5, and the molecular weights were found to range between 15,000 and 61,000.

Using a highly sensitive in vitrobioassay system, luteinizing hormone activity has been measured in parallel with radioimmunoassays in postmenopausal plasma and plasma obtained from women in various phases of the menstrual cycle (follicular, mid-cycle, luteal). Biological and immunological activities were measured directly in plasma samples without any chemical manipulation. The biological activity (B) was always higher than the immunological activity (I); the B/I ratio varied from 2.1 to 14.0. Gel filtration of pooled plasma samples through Sephadex G-100 revealed major discrepancies in each physiological state when immunological and biological activities were measured in each fraction. The biological activity was eluted as a single peak behind the elution volume of bovine serum albumin, but in front of the elution volume of chymotrypsinogen. It was invariably preceded by a small hump. The immunological activity was spread all over the chromatogram. Areas of immunological activity without any biological activity were located on either side of the biologically active fractions, both in the high molecular weight range (including the void volume) and in the low molecular weight range. The biological LH activity recovered following fractionation on Sephadex G-100 was in close agreement with that loaded, whereas the immunological activity recovered following gel filtration exceeded the loaded activity by a factor of 6–8. In the various physiological states, 11 to 44 % of the total immunological activity recovered was not associated with any biological activity. Furthermore, there was a marked variation in the ratio of biological to immunological activities of those fractions which contained biological activity.It is suggested that the specificity of current RIA methods could be improved significantly by preparing antisera which react only with biologically active LH species.

Summary.The in-vitro bioassay for LH, using mouse Leydig cells, has been modified for the direct measurement of serum LH in the male rhesus monkey. Validation of the assay demonstrated good reliability in terms of accuracy, precision and sensitivity (1·5 mi.u./ml). Basal LH concentrations in laboratory-maintained monkeys with and without anaesthesia were not significantly different from those in free-ranging, feral monkeys. LH-RH (50 μg i.v.) elicited a 30-fold increase in LH concentrations after 30 min. LH levels in castrated adult monkeys were approximately 50 times the normal levels. Intact and castrated juvenile males had only very low LH levels. LH from the serum of an adult male castrate was further characterized by Sephadex G100 column chromatography.The in-vitro bioassay provides a preferable alternative to the heterologous radioimmunoassay method for the routine determination of LH in the rhesus monkey.

Bulk hydrogels traditionally used for tissue engineering and drug delivery have numerous limitations, such as restricted injectability and a nanoscale porosity that reduces cell invasion and mass transport. An evolving approach to address these limitations is the fabrication of hydrogel microparticles (i.e., “microgels”) that can be assembled into granular hydrogels. There are numerous methods to fabricate microgels; however, the influence of the fabrication technique on granular hydrogel properties is unexplored. Herein, we investigated the influence of three microgel fabrication techniques (microfluidic devices (MD), batch emulsions (BE), and mechanical fragmentation by extrusion (EF)) on the resulting granular hydrogel properties (e.g., mechanics, porosity, and injectability). Hyaluronic acid (HA) modified with various reactive groups (i.e., norbornenes (NorHA), pentenoates (HA-PA), and methacrylates (MeHA)) were used to form microgels with an average diameter of ∼100 μm. The MD method resulted in homogeneous spherical microgels, the BE method resulted in heterogeneous spherical microgels, and the EF method resulted in heterogeneous polygonal microgels. Across the various reactive groups, microgels fabricated with the MD and BE methods had lower functional group consumption when compared to microgels fabricated with the EF method. When microgels were jammed into granular hydrogels, the storage modulus (G′) of EF granular hydrogels (∼1000–3000 Pa) was consistently an order of magnitude higher than G′ for MD and BE granular hydrogels (∼50–200 Pa). Void space was comparable across all groups, although EF granular hydrogels exhibited an increased number of pores and decreased average pore size when compared to MD and BE granular hydrogels. Furthermore, granular hydrogel properties were tuned by varying the amount of cross-linker used during microgel fabrication. Lastly, granular hydrogels were injectable across formulations due to their general shear-thinning and self-healing properties. Taken together, this work thoroughly characterizes the influence of the microgel fabrication technique on granular hydrogel properties to inform the design of future systems for biomedical applications.