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The aim of the study was to produce Ca2+ and Mg2+ independent raw starch hydrolyzing 1,4-α-d-glucan glucohydrolase (GGH) for glucose and fructose syrup production. A thermophilic GGH was produced from Aspergillus oryzae NRRL 1560. Two iso-forms of the enzyme, i.e., GGH1 and GGH2, were purified to homogeneity up to 4.93- and 8.82-fold that showed broad bands of 40 and 48 kDa on SDS-PAGE, respectively. Km and Vmax values of GGH2 were 1.315 mg mL−1 and 70.864 μmol mL−1 min−1, respectively. Enzyme showed optimum activity at 60 °C, pH 5.0. It displayed a broad range of pH (3.0–9.0) and temperature (30–60 °C) stability. Inactivation kinetics parameters were calculated as Kd = 0.014 min−1, t1/2 = 49.5 min, D = 164.5 min, z = 22.23 °C, Tm = 77 °C, and Ea(d) = 337.481 kJ/mol. Thermodynamics parameters for thermal unfolding were ΔG*(d) = 264.323, ΔH*(d) = 334.713 kJ/mol, and ΔS*(d) = 0.211 kJ/mol K. GGH2 was activated by 1 mM Na+ (18%), K+ (15%), Mg2+ (6%), Ca2+ (1%) and inhibited by Cu2+ (21%), Zn2+ (10%), Mn2+ (6%) at the same concentrations. The enzyme was also inhibited by 5 mM SDS (95%), DTT (8%), β-mercaptoethanol (4%), and EDTA (2%). GGH2-hydrolyzed raw starches were in the following order rice > wheat > corn > sweet potato > potato. This is the first report on raw starch digesting enzyme from A. oryzae. All these features make this enzyme a potential candidate for industrial production of glucose/fructose syrups and ethanol from raw starch via biological conversion.

Purpose: To find out the effect of suprachoroidal Triamcinolone injection on best corrected visual acuity (BCVA) and central retinal thickness (CRT) in patients with macular edema secondary to retinal vein occlusion. Study Design: Interventional case series. Place and Duration of Study: College of Ophthalmology and Allied vision Sciences, Ophthalmology department, Unit II Mayo hospital, Lahore, from September 2019 to January 2020. Methods: This study included 45 patients diagnosed with unilateral, retinal vein occlusion associated with macular edema. Patients with previous anti-vascular endothelial growth factor injection or any steroid injection received in the last 3 months or macular edema due to any other cause were excluded from the study. Only one eye of each patient was enrolled. The patients were treated with suprachoroidal triamcinolone injection (4 mg/100µL concentration). Patients with baseline central retinal thickness (CRT) of > 300 µm were included in the study. Serial changes in this parameter were evaluated at 1 week, 1 month and 3?months after suprachoroidal triamcinolone injection. Final CRT and Best-corrected visual acuity (BCVA) was recorded after three months. Results: Out of 45 patients, 26 (57.7%) were males and 19 (42.2%) were females. Majority of the patients (35.4%) were 51–60 years old. During first week the visual acuity was 0.321 ± 0.273 LogMAR, after one month it was 0.468 ± 0.291 and 0.406 ± 0.318 after 03 months with a p value of 0.003. After three months significant decrease in CRT was observed. With a p-value of 0.002. Conclusions: Suprachoroidal injection significantly improves BCVA and decreases CRT in patients with macular edema due to retinal vein Occlusion. Keywords: Retinal vein occlusion, suprachoroidal injection, triamcinolone acetonide.

Internet of Things (IoT) is rapidly increasing day by day due to its involvement in many applications such as electric grids, biological networks, transport networks, etc. In complex network theory, the model based on Scale Free Networks (SFNs) is more suitable for IoT. The SFNs are robust against random attacks; however, vulnerable to malicious attacks. Furthermore, as the size of a network increases, its robustness decreases. Therefore, in this paper, we propose a novel topology evolution approach to enhance the robustness of SFNs. Initially, we divide the network area into upper and lower parts. The nodes are deployed equally in both parts and connected via one-to-many correspondence. The distribution is made because small sized networks are more robust against malicious attacks. Moreover, we use k-core decomposition to calculate the hierarchical changes in the nodes’ degree. In addition, the core-based and degree-based attacks are performed to analyze the robustness of SFNs. For the network optimization, we compare the Genetic Algorithm (GA) with Artificial Bee Colony (ABC) and Bacterial Foraging Algorithm (BFA). In the optimization process, the node’s distance based edge swap is performed to draw long links in the network because these links make the network more robust.

Yeast extract is extensively applied in various food industries as a food additive to enhance to flavor of food products or as a vitamin supplement. It is also considered as a crucial component of microbiological media. The current study was conducted to optimize a process for the production of yeast extract by using Baker’s yeast (Saccharomyces cerevisiae). The cultivation of yeast biomass was performed in a stirred fermenter. The influence of numerous physical and chemical parameters such as carbon and nitrogen sources, temperature, pH and agitation were evaluated on the production of yeast cells by employing one factor at a time approach and optimum conditions for the production of maximum yeast biomass was determined. The maximum growth was attained using molasses as a substrate at 30ºC supplemented with urea at 150 rpm with pH 4.5. After fermentation, cells were separated by centrifugation and were ruptured by adopting different techniques and autolysis was found the most viable method. Various techniques were applied to dry the yeast extract and the spray dryer was appeared as most effective one. Yeast extract acquired after drying was subjected to various analysis including protein and solid content estimation and amino acid profiling and compared with commercial yeast extract. The dried yeast extract was incorporated in media preparations to grow various microorganisms including yeast, fungi and bacteria and considerable growth was observed. These promising results indicated that the developed process is a cost effective alternative approach for the production of yeast extract.

Microbial fuel cell (MFC) technology is an emerging area for alternative renewable energy generation and it offers additional opportunities for environmental bioremediation. Recent scientific studies have focused on MFC reactor design as well as reactor operations to increase energy output. The advancement in alternative MFC models and their performance in recent years reflect the interests of scientific community to exploit this technology for wider practical applications and environmental benefit. This is reflected in the diversity of the substrates available for use in MFCs at an economically viable level. This review provides an overview of the commonly used MFC designs and materials along with the basic operating parameters that have been developed in recent years. Still, many limitations and challenges exist for MFC development that needs to be further addressed to make them economically feasible for general use. These include continued improvements in fuel cell design and efficiency as well scale-up with economically practical applications tailored to local needs.

The production of lovastatin by Aspergillus terreus PU-PCSIR-1 was studied using submerged fermentation technique. Effects of pH, incubation temperature, incubation time, and the media components on the production of lovastatin were investigated. Maximum lovastatin yield (198.90 mg/L) was achieved after 14 days of incubation at temperature 28°C and pH 7.4. Peptonized milk, linoleic acid, glucose, yeast extract, and trace elements had a positive impact on the production of lovastatin by A. terreus.

Cell lysate of Escherichia coli strain BL21 showed significant D-glucose isomerase activity. The rate of glucose conversion was increased up to 40% when cells were induced with 1% D-xylose. E. coli BL21 xylose isomerase (ECXI-BL21) was purified to homogeneity, up to 1.9-fold with overall 10.88% enzyme yield by heat shock, salting out and electro-elution. The molecular mass of ECXI-BL21 was estimated as 43.9 kDa on SDS-PAGE. pH(opt.) and T(opt.) of the enzyme were calculated as 7.0 and 50 °C, respectively. Activation energy (E(a)) of ECXI-BL21 was 45 kJ/mol. Enzyme was stable from 30 to 55 °C and at pH range 6.0–8.0. ECXI-BL21((holo)) was activated by 10 mM magnesium (35%), 0.5 mM cobalt (20%) and manganese (25%), and 0.5/10 mM Mn(2+)/Mg(2+) (50%) and Co(2+)/Mg(2+) (30%) as compared to ECXI-BL21((apo)). Catalytic affinity (K(m)) of ECXI-BL21 for D-glucose was calculated as 0.82 mM, while maximum velocity (V(max)) of the reaction D-glucose((aldo)) ⇌ D-fructose((keto)) was 108 μmol/mg/min. D-fructose formed was identified on silica gel plate. This thermophilic enzyme, T(m) = 75 °C, has great potential for high fructose syrup production used in food and soft drink industries.

Prostate cancer (PC) is the second most common malignancy in men. According to International agency for research on cancer,4552 new cases of PC were registered in 2018 in Pakistan.Although majority of cases are confined to prostate on initial presentation, there is a high tendency for advanced PC to metastasize to bone. Metastatic lesions are typically osteoblastic rather than osteolytic. Therefore, these are easily identified by Technetium labeled Methylene Diphosphonate (Tc99m-MDP) uptake on Whole body Bone Scan (BS). Hybrid scanning offers anatomic details for differentiation between aggressive or non-aggressive lesions. Most common axial sites include pelvic bones and spine. Metastases to appendicular skeleton is rare and uncommon . We present a case of 62-years-old male with PC. Follow-up WBS, showed interval development of multiple sites of skeletal metastases. SPECT-CT scan acquired for characterization of atypical site of abnormal MDP uptake in appendicular skeleton, which showed features suggesting skeletal metastasis.

Blockchain is a decentralized and shared dis-tributed ledger that records the transaction history done by totally different nodes within the whole network. The technology is practically used in the field of education for record-keeping, digital certification, etc. There have already been several papers published on this, but no one can’t find a single paper covering the blockchain-based educational projects. So, There is a gap of latest trends to education. Blockchain-based educational projects resolve the issues of today’s educators. On that basis, we conclude that there is a need for conducting a systematic literature review. This study, therefore, reviews the artistic gap between these two based on educational projects. For this purpose, the paper focuses on exploring some block-chain based projects and protocols that are used in these projects. It also analyses the block-chain features that are being used and the services are offered by the existing educational projects using block-chain features to improve the execution of this technology in education.

A thermophilic bacterium (TP-2) was isolated from the Tatta Pani hot spring in Azad Kashmir and was characterized using phenotypic and genotypic characters. The strain developed cream colored, round, smooth, flat and slimy colonies while the cells were Gram positive rods that ranged in size from about 2.1-3.6 μm to 0.2-0.3 μm in width. Sequence analysis of its 16S rRNA gene showed that isolate TP-2 had 89% homology with Geobacillus debilis. It grew within pH range of 5.5 to 8.5 with optimum growth at pH 7.0. The isolate showed optimum growth at 65ºC and gave positive results for gelatin hydrolysis (GEL), ortho nitrophenyl-β-D-galactopyranosidase (ONPG), and nitrate production and produced acid from sucrose, glucose and maltose. It utilized glucose, fructose, maltose, lactose, sucrose, xylan, starch, filter paper and carboxymethylcellulose as sole carbon source. Isolate TP-2 produced significant amount of industrially important enzymes i.e. extracellular α-amylase, CMCase, FPase, Xylanase, Protease and Lipase and intracellular CMCase and FPase.

A genomic DNA fragment, encoding a thermotolerant β-glucosidase, of the obligate anaerobe Thermotoga petrophila RKU-1 was cloned after PCR amplification into Escherichia coli strain BL21 CodonPlus. The purified cloned enzyme was a monomeric, 51.5 kDa protein (by SDS-PAGE) encoded by 1.341 kb gene. The estimated K m and V max values against p-nitrophenyl-β-D-glucopyranoside were 2.8 mM and 42.7 mmol min−1 mg−1, respectively. The enzyme was also active against other p-nitrophenyl substrates. Possible catalytic sites involved in hydrolyzing different p-nitrophenyl substrates are proposed based on docking studies of enzyme with its substrates. Because of its unique characters, this enzyme is a potential candidate for industrial applications.

In this paper, we propose a lexical routing metric to enable path-wise link quality-aware routing in Wireless Sensor Networks (WSNs). The realization of this routing metric is achieved by applying the indexing techniques of formal language processing in multi-metric route classification and cost evaluation for WSNs. In particular, the metric is motivated from the fact that IEEE 802.15.4 networks are formed on links with highly variable quality, and selection of poor quality links degrades the data delivery considerably. Although IEEE 802.15.4 supports link-level awareness through LQI, (a) it remains unbeknownst at the path level, and therefore, (b) its processing is link-local in scope. We propose LABILE, a composite routing metric, that is implemented through modifying RREQ and RREP structures of AODV to capture and convey two-state link information to destination which in turn is processed through an easy to compute routing lexicon and a corresponding lexical algorithm for path selection in case of availability of multiple paths. Multiple paths are represented in a path space through a proposed cost model that encompasses hop-count, weak links and a quantity weakness factor of each link, depending upon a thresholding mechanism which in turn declares a link to be either healthy (aka usable) or weak (aka unusable). Using the weakness factor, the success probability of data delivery over weak link turns out to be a fraction of success probability at healthy link. The mathematical model along with the experiments done for LABILE show that proposed composite metric and its parsing scheme together achieve link robustness consistently by evading the link failures, as the number of weak links is fairly reduced in lexical path as compared to the hop-count-based metric. Increased data delivery is obtained through preventing retransmissions as on failure-prone links.

A 1.4 Kb fragment of Bacillus licheniformis ATCC 14580 encoding β-glucosidase was cloned and expressed in Escherichia coli. β-Glucosidase expressed by E. coli harboring cloned gene was located entirely in the intracellular fraction. Recombinant β-glucosidase protein was purified to homogeneity level and the molecular weight was found to be 53 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. It gave maximum activity at 50°C and pH 6. K m and V max were 0.206 mM and 1.26 U/mg, respectively, with p-nitrophenyl-β-D-glucopyranoside, while activation energy Ea, enthalpy of activation ?H and entropy of activation ΔS were found to be 66.31 kJ/mol, 64.04 kJ/mol and 48.28 J/mol/K, respectively. The pKa1 and pKa2 of the ionizable groups of active site residues involved in Vmax were found to be 5.5 and 7.0, respectively. When the recombinant β-glucosidase protein was used as a member of consortium with endoglucanase and exoglucanase for the saccharification of wheat straw, 123% increase in saccharification was observed.

This paper evaluates the genetic diversity of goat breeds in Pakistan, a country blessed with a wide range of goats spread throughout with distinct features contributing to a rich Animal Genetic Resource. The genetic diversity of two goat breeds (Jattal and Dera Din Panah) was assessed with 25 animals representing the two breeds using 16 microsatellite markers. The mean observed and expected heterozygosity of both goat breed populations were observed as 0.83+0.21. The average number of observed alleles was 3.6+1.6 for all loci. The mean polymorphic information content for a goat breed was 0.45, indicating the usefulness of markers panel. Highest Nei’s standard genetic distance (Ds) value of 0.0612 was observed between Jattal and Dera Din Panah goats, and the mean Fst value was 0.013. The measures of genetic variation revealed a good scope for effective improvement, conservation, and designing national breeding policies, in future, for Pakistan goat breeds.

Inorganic storage granules have long been recognized in bacterial and eukaryotic cells but were only recently identified in archaeal cells. Here, we report the cellular organization and chemical compositions of storage granules in the Euryarchaeon, Archaeoglobus fulgidusstrain VC16, a hyperthermophilic, anaerobic, and sulfate-reducing microorganism. Dense granules were apparent inA. fulgiduscells imaged by cryo electron microscopy (cryoEM) but not so by negative stain electron microscopy. Cryo electron tomography (cryoET) revealed that each cell contains one to several dense granules located near the cell membrane. Energy dispersive X-ray (EDX) spectroscopy and scanning transmission electron microscopy (STEM) show that, surprisingly, each cell contains not just one but often two types of granules with different elemental compositions. One type, named iron sulfide body (ISB), is composed mainly of the elements iron and sulfur plus copper; and the other one, called polyphosphate body (PPB), is composed of phosphorus and oxygen plus magnesium, calcium, and aluminum. PPBs are likely used for energy storage and/or metal sequestration/detoxification. ISBs could result from the reduction of sulfate to sulfide via anaerobic energy harvesting pathways and may be associated with energy and/or metal storage or detoxification. The exceptional ability of these archaeal cells to sequester different elements may have novel bioengineering applications.

Mercury is present in different types of industrial effluents that cause environmental pollution. Conventional methods such as precipitation, oxidation/reduction, ion exchange, filtration, membranes, and evaporation are extremely expensive or inefficient for the removal of mercury from diluted solutions. In this context, the biosorption process has recently been shown to be an effective and economical method. The present work describes the mercury biosorption ability of three fungi, i.e., Aspergillus niger, Trichoderma viride, and Humicola insolens. Monocultures of these strains and 10 different combinations were investigated. The consortium of 24-h-old H. insolens and 48-h-old of A. niger and T. viride in equal ratio was found to be compatible. This consortium decreased the residual mercury from 2.02 to 0.001 μ g/L after 7 days of incubation, and caused a significant reduction in chemical oxygen demand (COD) (92.6%) from an initial level of 21 mg/L.

Lung cancer is one of the most prevalent malignancies in the world and both incidence and mortality rates are continuing to rise in Pakistan. However, epidemiological studies to identify common lung cancer determinants in the Pakistani population have been limited.In this retrospective case-control study, 400 cases and 800 controls were enrolled from different hospitals of all provinces of Pakistan. Information about socio-demographic, occupational, lifestyle and dietary variables was extracted by questionnaire from all subjects. Odd ratios (ORs) and 95% confidence intervals (CIs) were calculated. and dose-response associations were also assessed for suitable factors.Strong associations were observed for smoking (OR=9.4, 95%CI=6.9-12.8), pesticide exposure (OR=5.1, 95%CI=3.1-8.3), exposure to diesel exhaust (OR=3.1, 95%CI=2.1-4.5), red meat consumption (OR=2.9, 95%CI=1.8-4.7) and chicken consumption (OR=2.8, 95%CI=1.7-49). Other associated factors observed were welding fumes (OR=2.5, 95%CI=1.0-6.5), sedentary living (OR=2.0, 95%CI=1.6-2.6), family history (OR=2.0, 95%CI=0.8-4.9), wood dust (OR=1.9, 95%CI=1.2- 3.1), tea consumption (OR=1.8, 95%CI=1.2-2.6), coffee consumption (OR=1.8, 95%CI=1.1-2.8), alcoholism (OR=1.7, 95%CI=1.1-2.5) and asbestos exposure(OR=1.5, 95%CI=0.5-4.4). Consumption of vegetables (OR=0.3, 95%CI=0.2-0.4), juices (OR=0.3, 95%CI=0.3-0.4), fruits (OR=0.7, 95%CI=0.5-0.9) and milk (OR=0.6, 95%CI=0.5- 0.8) showed reduction in risk of lung cancer. Strongest dose-response relationships were observed for smoking (?2=333.8, p≤0.0000001), pesticide exposure (?2=50.9, p≤0.0000001) and exposure to diesel exhaust (?2=51.8, p≤0.0000001).Smoking, pesticide exposure, diesel exhaust and meat consumption are main lung cancer determinants in Pakistan. Consuming vegetables, fruits, milk and juices can reduce the risk of lung cancer risk, as in other countries.

The production of an extracellular β-glucosidase by Aspergillus niger NRRL 599 was optimized using submerged fermentation technique. Effect of different media, different carbon sources, initial pH of the fermentation medium, temperature, incubation period and inoculum size on the production of β- glucosidase enzyme was investigated. A. niger NRRL 599 produced maximum extracellular β- glucosidase (4.48 U/mg) in Eggins and Pugh medium with 1% wheat bran (w/v) at pH 5.5 inoculated with 4% conidial suspension after 96 h of incubation at 30°C. Purified β-glucosidase gave K m and V max values of 3.11 mM and 20.83 U/mg respectively for pNPG hydrolysis. The enzyme was optimally active at pH 4.8 and at temperature of 60°C. Thermodynamic parameters, E a , ΔH and ΔS were found to be 52.17 KJ/mol, 49.90 J/mol.K and -71.69 KJ/mol, respectively. The pKa1 and pKa 2 of ionizable groups of active site residues involved in V max were calculated to be 4.1 and 6.0 respectively. Key words : β-Glucosidase, Aspergillus niger, kinetics, thermodynamics.

An endo-1,4-β-glucanase gene from moderate thermophilic strain Bacillus licheniformis ATCC 14580 was cloned into Escherichia coli DH5α using pTZ57R/T as cloning vector. After confirmation of clone through polymerase chain reaction (PCR) and restriction analysis, endoglucanase gene (1.5 kb) was further expressed in E. coliBL21 (DE 3) using pET-22b (+) expression vector. The molecular weight of endoglucanase gene was found to be 52.2 kDa as determined by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The addition ofisopropyl-β-D-thio-galactoside (IPTG) as an inducer in the fermentation medium, caused an increase in expression of recombinant endo-glucanase. The activity of enzyme in optimized condition was found to be 1.5 U/ml using carboxymethylcellulose as substrate. The optimum pH and the temperature of enzyme were also determined and found to be 6.0 and 60°C, respectively. Key words: Endo-1,4-β-glucanase, Bacillus licheniformis, cloning, expression.

Wheat bran, a by-product of wheat milling industry, is the outermost covering of wheat grain. It has relatively diverse application in food, feed, medicine and fermentation industries due to its richness in carbohydrates (mostly fibers), protein and fats which makes it an important dietary element. Its importance from medical view point is well documented especially in reducing blood plasma cholesterol and prevention of cardiovascular diseases and certain cancers, that is, colon cancer. In fermentation industry, wheat bran can be used as a substrate for production of a range of products such as biomass, enzyme, biofuel and production of other metabolites. In short wheat bran has immense applications and market value which makes it so important to be considered as brown gold. Key words: Wheat bran, enzyme production, biofuel, food, health.

2, 9, Walailak Journal of Science and Technology

In this paper, we propose a framework to apply the indexing techniques of formal language processing in multi-metric route classification and cost evaluation for WSNs. Specifically, we present an easy to compute, dynamic routing lexicon and a corresponding lexicographical algorithm for path selection in case of availability of multiple paths. The cost evaluation of paths and path selection in case of availability of multiple paths is simple and particularly appealing for devices having low power and limited computational capabilities and other impairments. The proposed lexical approach has the potential to be applied over a myriad of WSNs. The benefits of our lexicographical approach over the existing weighted sum approach are particularly asserted. However, the proposed algorithm makes rigid decisions based on the preferred metric regardless of costs incurred by other metrics; we therefore present at the end a plausible criterion to remove this rigidity in path selection.

Scrapie is a fatal neurodegenerative disorder of sheep and goats caused by post-translational conformational change in the host-encoded prion protein (PrPC). Susceptibility or resistance to scrapie has been associated with the presence of polymorphisms in the prion protein (PrP) gene. In the present study, we analyzed the PrP gene sequence to determine the frequency of polymorphisms in 56 sheep (28 each from Damani and Hashtnagri breeds) and 56 goats (28 each from Kamori and Local Hairy breeds). A total of 7 amino acid polymorphisms were detected in the PrP gene for sheep and 4 for goats. These amino acid polymorphisms were combined in 13 alleles and 15 genotypes in sheep and 5 alleles and 6 genotypes in goats. The overall frequency of the most sheep scrapie-resistant polymorphism (Q171R) was calculated to be 0.107. The most scrapie-susceptible polymorphism (A136V) was not detected in any of the studied sheep. The overall frequency of scrapie-associated polymorphism (H143R) in goats was found to be 0.152. Along with already known amino acid polymorphisms, two novel polymorphisms were also detected for each of sheep (Q171N and T191I) and goats (G22C and P63L). However, the overall frequency of these polymorphisms was extremely low.