An endo-acting proline-specific oligopeptidase from Treponema denticola ATCC 35405: evidence of hydrolysis of human bioactive peptides

An endo-acting proline-specific oligopeptidase (prolyl oligopeptidase [POPase], EC 3.4.21.26) was purified to homogeneity from the Triton X-100 extracts of cells of Treponema denticola ATCC 35405 (a human oral spirochete) by a procedure that comprised five successive fast protein liquid chromatography steps. The POPase is a cell-associated 75- to 77-kDa protein with an isoelectric point of ca. 6.5. The enzyme hydrolyzed (optimum pH 6.5) the Pro-pNA bond in carbobenzoxy-Gly-Pro-p-nitroanilide (Z-Gly-Pro-pNA) and bonds at the carboxyl side of proline in several human bioactive peptides, such as bradykinin, substance P, neurotensin, angiotensins, oxytocin, vasopressin, and human endothelin fragment 22-38. The minimum hydrolyzable peptide size was tetrapeptide P3P2P1P'1, while the maximum substrate size was ca. 3 kDa. An imino acid residue in position P1 was absolutely necessary. The hydrolysis of Z-Gly-Pro-pNA was potently inhibited by the following, with the Ki(app) (in micromolar) in parentheses: insulin B-chain (0.7), human endothelin-1 (0.5), neuropeptide Y (1.7), substance P (32.0), T-kinin (4.0), neurotensin (5.0), and bradykinin (16.0). Chemical modification and inhibition studies suggest that the POPase is a serine endopeptidase whose activity depends on the catalytic triad of COOH ... Ser ... His but not on a metal. The amino acid sequence around the putative active-site serine is Gly-Gly-Ser-Asn-Pro-Gly. The enzyme is suggested to contain a reactive cysteinyl residue near the active site. Amino acid residues 4 to 24 of the first 24 N-terminal residues showed a homology of 71% with the POPase precursor from Flavobacterium meningosepticum and considerable homology with the Aeromonas hydrophila POPase. The ready hydrolysis of human bioactive peptides at bonds involving an imino acid residue suggests that enzymes like POPase may contribute to the chronicity of periodontal infections by participating in the peptidolytic processing of those peptides.     

SPARC deficiency leads to early-onset cataractogenesis

PURPOSE: To determine the role of SPARC (secreted protein, acidic, and rich in cysteine) in cataractogenesis by examining mice deficient in a matricellular protein SPARC. METHODS: Mice were rendered SPARC-deficient by a targeted disruption of the gene. Slit-lamp microscopy and histology were used to examine the eyes of SPARC-null and wild-type mice from birth to 14 months of age. RESULTS: SPARC-null mice developed opacities in the posterior cortex of the eye as early as 1.5 months after birth. The diffuse cataracts appeared to progress toward the anterior cortex and reached maturity in many animals by 3.5 months of age. Early stages of cataractogenesis in SPARC-null mice included inhibition of normal lens fiber cell differentiation, degeneration of fiber cells, vacuole formation at the equator, and liquefaction of the cortex. No cataracts were detected in wild-type mice up to the age of 8 months. CONCLUSIONS: The early onset of cataracts in SPARC-null mice establishes that the gene is essential to the maintenance of lens transparency.     

Lung nodules detection using semantic segmentation and classification with optimal features

Neural Computing and Applications - Lung cancer is a deadly disease if not diagnosed in its early stages. However, early detection of lung cancer is a challenging task due to the shape and size of...     

Current Understandings on Magnesium Deficiency and Future Outlooks for Sustainable Agriculture

The productivity of agricultural produce is fairly dependent on the availability of nutrients and efficient use. Magnesium (Mg²⁺) is an essential macronutrient of living cells and is the second most prevalent free divalent cation in plants. Mg²⁺ plays a role in several physiological processes that support plant growth and development. However, it has been largely forgotten in fertilization management strategies to increase crop production, which leads to severe reductions in plant growth and yield. In this review, we discuss how the Mg²⁺ shortage induces several responses in plants at different levels: morphological, physiological, biochemical and molecular. Additionally, the Mg²⁺ uptake and transport mechanisms in different cellular organelles and the role of Mg²⁺ transporters in regulating Mg²⁺ homeostasis are also discussed. Overall, in this review, we critically summarize the available information about the responses of Mg deficiency on plant growth and development, which would facilitate plant scientists to create Mg²⁺-deficiency-resilient crops through agronomic and genetic biofortification.     

Experimental and Established Oximes as Pretreatment before Acute Exposure to Azinphos-Methyl

Poisoning with organophosphorus compounds (OPCs) represents an ongoing threat to civilians and rescue personal. We have previously shown that oximes, when administered prophylactically before exposure to the OPC paraoxon, are able to protect from its toxic effects. In the present study, we have assessed to what degree experimental (K-27; K-48; K-53; K-74; K-75) or established oximes (pralidoxime, obidoxime), when given as pretreatment at an equitoxic dosage of 25% of LD₀₁, are able to reduce mortality induced by the OPC azinphos-methyl. Their efficacy was compared with that of pyridostigmine, the only FDA-approved substance for such prophylaxis. Efficacy was quantified in rats by Cox analysis, calculating the relative risk of death (RR), with RR=1 for the reference group given only azinphos-methyl, but no prophylaxis. All tested compounds significantly (p ≤ 0.05) reduced azinphos-methyl-induced mortality. In addition, the efficacy of all tested experimental and established oximes except K-53 was significantly superior to the FDA-approved compound pyridostigmine. Best protection was observed for the oximes K-48 (RR = 0.20), K-27 (RR = 0.23), and obidoxime (RR = 0.21), which were significantly more efficacious than pralidoxime and pyridostigmine. The second-best group of prophylactic compounds consisted of K-74 (RR = 0.26), K-75 (RR = 0.35) and pralidoxime (RR = 0.37), which were significantly more efficacious than pyridostigmine. Pretreatment with K-53 (RR = 0.37) and pyridostigmine (RR = 0.52) was the least efficacious. Our present data, together with previous results on other OPCs, indicate that the experimental oximes K-27 and K-48 are very promising pretreatment compounds. When penetration into the brain is undesirable, obidoxime is the most efficacious prophylactic agent already approved for clinical use.     

A Novel DUF569 Gene Is a Positive Regulator of the Drought Stress Response in Arabidopsis

In the last two decades, global environmental change has increased abiotic stress on plants and severely affected crops. For example, drought stress is a serious abiotic stress that rapidly and substantially alters the morphological, physiological, and molecular responses of plants. In Arabidopsis, several drought-responsive genes have been identified; however, the underlying molecular mechanism of drought tolerance in plants remains largely unclear. Here, we report that the “domain of unknown function” novel gene DUF569 (AT1G69890) positively regulates drought stress in Arabidopsis. The Arabidopsis loss-of-function mutant atduf569 showed significant sensitivity to drought stress, i.e., severe wilting at the rosette-leaf stage after water was withheld for 3 days. Importantly, the mutant plant did not recover after rewatering, unlike wild-type (WT) plants. In addition, atduf569 plants showed significantly lower abscisic acid accumulation under optimal and drought-stress conditions, as well as significantly higher electrolyte leakage when compared with WT Col-0 plants. Spectrophotometric analyses also indicated a significantly lower accumulation of polyphenols, flavonoids, carotenoids, and chlorophylls in atduf569 mutant plants. Overall, our results suggest that novel DUF569 is a positive regulator of the response to drought in Arabidopsis.     

Tensile Stress Relaxation of Alumina at High Temperature

The development of a tensile testing methodology for ceramics which enables a stress vs strain-rate response to be measured at high temperature is described. The test involves a carefully controled stress relaxation test at constant total strain using an experimental procedure and phenomenological analysis previously developed for metallic materials. It is demonstrated here with preliminary tests on alumina at 1050° and 1150°C. This offers, with further development, the possibility of establishing design stresses associated with low strain-rate behavior for structural applications. The results demonstrate that data covering four decades of strain rate may be generated in tests lasting a few hours. The inelastic strain consists of substantial anelastic recoverable strain in addition to a permanent creep strain.     

Identification of probe request attacks in WLANs using neural networks

Any sniffer can see the information sent through unprotected ‘probe request messages’ and ‘probe response messages’ in wireless local area networks (WLAN). A station (STA) can send probe requests to trigger probe responses by simply spoofing a genuine media access control (MAC) address to deceive access point (AP) controlled access list. Adversaries exploit these weaknesses to flood APs with probe requests, which can generate a denial of service (DoS) to genuine STAs. The research examines traffic of a WLAN using supervised feed-forward neural network classifier to identify genuine frames from rogue frames. The novel feature of this approach is to capture the genuine user and attacker training data separately and label them prior to training without network administrator’s intervention. The model’s performance is validated using self-consistency and fivefold cross-validation tests. The simulation is comprehensive and takes into account the real-world environment. The results show that this approach detects probe request attacks extremely well. This solution also detects an attack during an early stage of the communication, so that it can prevent any other attacks when an adversary contemplates to start breaking into the network.     

Novel water sorption behavior of emulsion blends

As waterborne emulsions continue to replace their solvent‐based counterparts in various applications, the water resistance of cast films is attracting significant interest. The characterization of the water sorption kinetics and structure–property relationships related to water sorption is, therefore, an important area for investigation. In this study, the water sorption kinetics of emulsion blends were compared with those of their blend components. The investigated blends were at equal weight (dry basis) fractions for each emulsion. The initial water sorption rate for immiscible emulsion blends was found to be significantly higher than composite values of the constituents. This behavior was due to percolation networks in the blends because thermodynamic constraints prevented diffusion across the interface bordering dissimilar particles, leaving a interface enriched with water‐sensitive species. The peak water sorption for the immiscible emulsion blends was lower than the composite values because of the ability of the water‐sensitive species to rapidly diffuse out of the samples due to the percolation network. This behavior existed for room‐temperature‐cast samples and persisted as the time and temperature exposure was increased. Atomic force microscopy results clearly showed the potential for percolation networks in the blends. Higher glass‐transition emulsion polymers [e.g., poly(vinyl acetate)] exhibited similar behavior, and this indicated poor film formation like that for the immiscible emulsion blends. These results indicated that the degree of film formation was critical with respect to the water sorption characteristics of emulsion films. Immiscible emulsion blends were compared with miscible emulsion blends for which all constituents exhibited excellent film formation (unblended). The immiscible blends exhibited a significant difference in water sorption compared with the miscible blends because of the existence of percolation networks. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 933–939, 2003     

KHATT: An open Arabic offline handwritten text database

A comprehensive Arabic handwritten text database is an essential resource for Arabic handwritten text recognition research. This is especially true due to the lack of such database for Arabic handwritten text. In this paper, we report our comprehensive Arabic offline Handwritten Text database (KHATT) consisting of 1000 handwritten forms written by 1000 distinct writers from different countries. The forms were scanned at 200, 300, and 600 dpi resolutions. The database contains 2000 randomly selected paragraphs from 46 sources, 2000 minimal text paragraph covering all the shapes of Arabic characters, and optionally written paragraphs on open subjects. The 2000 random text paragraphs consist of 9327 lines. The database forms were randomly divided into 70%, 15%, and 15% sets for training, testing, and verification, respectively. This enables researchers to use the database and compare their results. A formal verification procedure is implemented to align the handwritten text with its ground truth at the form, paragraph and line levels. The verified ground truth database contains meta-data describing the written text at the page, paragraph, and line levels in text and XML formats. Tools to extract paragraphs from pages and segment paragraphs into lines are developed. In addition we are presenting our experimental results on the database using two classifiers, viz. Hidden Markov Models (HMM) and our novel syntactic classifier.