Post-process optimization of 3D printed poly(lactic-co-glycolic acid) dental implant scaffold for enhanced structure and mechanical properties: effects of sonication duration and power.

We described a technique of a post-process stage to partially remove the poly(vinyl alcohol) (PVA) binder in Poly(lactic-co-glycolic acid) (PLGA) dental scaffolds. The scaffolds were exposed to ultrasonic waves while immersed in an ethanol/acetone solvent mixture that possessed both polar and nonpolar properties. A factorial experiment was conducted in which the scaffolds were treated to three levels of sonication power (pW): 0, 20% (22 W) and 40% (44 W), and soaking duration (t): 5, 15, and 30 min. The treated scaffolds were characterized by FT-IR, optical microscopy, and mechanical (compressive) testing. FT-IR revealed that the amount of PVA decreased with increasing pW and t. Two-way ANOVA revealed that increasing pW and t, respectively, resulted in increasing scaffold surface area to volume (SVR). Sonication and solvent caused structural damage (i.e., unevenness) on the scaffold surface, but the damage was minimal at 20% pW and 30 min. The optimal values of pW and t resulting in enhanced fracture strength, strain and toughness were 20% and 30 min, respectively, which corroborated the findings of minimal structural damage. However, sonication had no significant effects on the scaffold stiffness. Mechanistic analysis of the effects of sonication predicted that the ultrasonic energy absorbed by the scaffold was sufficient to disrupt the van Der Waals bonds between the PVA and PLGA but not high enough to disrupt the covalent bonds within the PLGA. This technique is promising as it can partially remove the PVA from the scaffold, and mitigate problematic issues down the line, such as thermal degradation during sterilization, and undue delay/variability in biodegradation.

An Improved VLSI Design of the ALU Based FIR Filter for Biomedical Image Filtering Application.

AIM: FIR filter is the most widely used device in DSP applications, which is also applicable to integrate with image processing approaches. The ALU based FIR structure is applicable for various devices to increase the performance. The ALU design operation includes accumulation, subtraction, shifting, multiplication and filtering. Existing methods are designed with various multipliers like Wallace tree multiplier, DADDA multiplier, Vedic multiplier and adders like carry select adder, and carry look-ahead adder. OBJECTIVE: The main objective is to reduce the area, delay and power factors since optimum VLSI circuit is employed in this paper. By these adders and multipliers, operations are independently enabling main operations in DSP. The FIR filter is designed using a MAC unit with clock regenerative comparators. INTRODUCTION: In the field of the VLSI industry, the low power, reduced time, and area-efficient designs are mostly preferred for various applications. Adders and multipliers play a vital role in VLSI circuit designs. The recent electronics industry uses a digital filter for various real-time applications. This utilizes Finite Impulse Response (FIR) and Infinite Impulse Response (IIR) filters, here the FIR filter is most stable than IIR filter. This FIR filter indicates the impulse signal into finite form and it is used mainly in DSP processors for getting high-speed. In these two ALU and FIR circuits, the adders and multiplier block's usage is increased, it consumes much power. METHODS: The proposed research work uses the clock-gating technique for reducing power consumption. Here the latch-based clock gating technique provides an efficient result. XOR-based logic circuit reduces the design complexity and utilizes the less area. Carry save accumulator is a digital adder used for addition. It provides the two set of output, which is partial sum and carry output. The ripple carry adder uses a full adder circuit for its operation. It propagates the carry value in the last bit. In addition, the combination of CSA and RCA utilizes less area, high speed and provides the better throughput. In multiplier block, the booth multiplier algorithm is used with XOR-based logic. Here this proposed FIR filter is designed for performing image filtration of retina image. This process improves the better visualization approach inthe medical field. RESULTS: Thus, the proposed ALU based FIR filter with a latch-based clock gating technique is designed and analyzed with various parameters. Here the modified adders and multipliers are proposed for the efficiency of the system. The modified carry save adder is proposed with combining ripple carry adder logic for improving the adders' performance. The enhanced booth multiplier is designed using add and shift method for reducing the numberof stages to calculate the result. This process is applied to perform image processing of the retina image. After designing the ALU based FIR filter structure in the VLSI environment, the image is loaded on the MATLAB as the .png format, then it is converted into a hex file, which is read from the Xilinx to perform filtering the process. Then the 'dataout' is converted into a binary file to obtain the result of the filtering process. The enhanced booth multiplier reduces the delay by reducing the number of stages to calculate the result. Here the clock gating technique is proposed with the latch-based design for reducing the dynamic and clock power consumption. The number of adder's circuit in both ALU and FIR circuits is less since it improves the overall efficiency of the system. CONCLUSION: Thus, the proposed methodology concluded that the design and analysis of ALU based FIR filter for medical image processing give the efficient result on the way of achieving the factors such as power (Static & Dynamic), Delay (Path delay) area utilization, MSE and PSNR. Here the image processing of FIR results to MSE and PSNR values, which obtained a better result than the existing VLSI based image processing works. The latch-based clock gating circuit is connected with the proposed circuit, based on the gated clock signal, it optimizes the gated circuit of the whole design since it also reduces the error and provides the efficient power report. This proposed VLSI model is simulated using Xilinx ISE 14.5 and Modelsim synthesizes it; here with the help of MATLAB, with the adaptation of the 2018a tool, the image filtering was done.

Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina.

OBJECTIVES: An extracellular protease inhibitor (BTPI-301) of trypsin was purified and characterized from an isolate of Pseudomonas mendocina. RESULTS: BTPI-301was purified to homogeneity by (NH4)2SO4, precipitation, DEAE Sepharose and CNBr-activated Sepharose chromatography. Homogeneity was proved by native PAGE and SDS-PAGE. The intact molecular mass was 11567 Da by MALDI-TOF analysis. BTPI-301was a competitive inhibitor with a Ki of 3.5 × 10-10 M. It was stable and active at pH 4-12 and also at 4-90 °C for 1 h. Peptide mass fingerprinting by MALDI revealed that the BTPI-301 is a new inhibitor not reported so far with protease inhibitory activity. The pI of the inhibitor was 3.8. The stoichiometry of trypsin-BTPI-301 interaction is 1:1. The inhibitor was specific towards trypsin. CONCLUSION: A pH tolerant and thermostable protease inhibitor BTPI-301 active against trypsin was purified and characterized from P. mendocina that could be developed and used as biopreservative as well as biocontrol agent.

Predictive value of amplitude-integrated EEG (aEEG) after rescue hypothermic neuroprotection for hypoxic ischemic encephalopathy: a meta-analysis.

OBJECTIVE: Amplitude-integrated electroencephalography (aEEG) is a useful bedside tool in predicting the neurodevelopmental outcome after neonatal encephalopathy; however, the prognostic accuracy may be altered by rescue hypothermic neuroprotection. The objective of this study is to examine the prognostic accuracy of aEEG for predicting long-term neurodevelopmental outcomes in term newborn infants undergoing therapeutic hypothermia for neonatal encephalopathy. STUDY DESIGN: We examined all studies (Medline, Cumulative Index to Nursing and Allied Health Literature and the Cochrane Library; 2000 to 2014) comparing aEEG (6, 24, 48 or 72 h) in term encephalopathic babies undergoing therapeutic hypothermia, with neurodevelopmental outcome at 1 year or more. We extracted individual patient data from the eligible studies to calculate prognostic indices with exact confidence intervals (CIs). We considered continuous normal voltage as normal aEEG pattern and discontinuous normal voltage, burst suppression, flat trace and persistently low voltage as abnormal, and defined adverse outcome as death or moderate/severe disability at 1 year. RESULTS: We reviewed a total of 70 articles, 17 of which met the inclusion criteria. Eight studies were excluded and 9 studies (N=520) were included in the meta-analysis. The pooled sensitivity and specificity for an abnormal trace at 6 h of age to predict adverse outcome were 96% (95% CI 91 to 98%) and 39% (95% CI 32 to 46%). The diagnostic odds ratio of an abnormal trace was highest at 48 h (66.9 (95% CI 19.7, 227.2)). CONCLUSIONS: A persistantly abnormal aEEG at 48 h or more is associated with an adverse neurodevelopmal outcome. The positive prognostic value of 6 h aEEG is poor and good outcome may occur despite abnormal aEEG. Conversely, a normal 6 h aEEG has a good negative predictive value although do not exclude adverse outcomes.

Differential Sensitivity of Madin-darby Canine Kidney (MDCK) Cells to Epinephrine.

Catecholamines regulate a variety of cellular functions in the mammalian kidney. The present study was aimed to investigate the differential sensitivity of Madin-Darby Kidney Cells (MDCK cells) to epinephrine in a dose-dependent manner. The loss of adhesion and altered cell shape were observed in MDCK cells. The presence of apoptosis and necrosis were studied by the fluorescence microscope and Confocal Laser Scanning Microscope (CLSM). Scanning Electron Microscope (SEM) analysis showed several surface microvilli, and cells were rounded having ruffled and crenated surface. Agarose gel electrophoresis study showed the presence of smearing, which further confirms the occurrence of necrosis. The fluorescence staining study showed the increased reactive oxygen species (ROS) level. Up-regulation of p53, bax, and caspase 3 mRNA expressions was evidenced by quantitative PCR (qPCR). Caspase 3 activity was also increased in epinephrine treated cells. Our experimental results do not imply that the epinephrine should not be used in the clinical treatments. However, our results add a research note of caution on the possible cytotoxic effect of maximal doses of epinephrine over a prolonged time.

Lipid peroxidation and antioxidants status in human malignant and non-malignant thyroid tumours.

INTRODUCTION: Thyroid epithelial cells produce moderate amounts of reactive oxygen species that are physiologically required for thyroid hormone synthesis. Nevertheless, when they are produced in excessive amounts, they may become toxic. OBJECTIVE: The present study is aimed to compare the lipid peroxidation (LPO), antioxidant enzymes - superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and non-protein thiols (reduced glutathione (GSH)) in human thyroid tissues with malignant and non-malignant disorders. DESIGN AND METHODS: The study used human thyroid tissues and blood samples from 157 women (147 diseased and 10 normal). Thyroid hormones, oxidative stress markers and antioxidants were estimated by standard methods. RESULTS: LPO significantly increased in most of the papillary thyroid carcinoma (PTC: 82.9%) and follicular thyroid adenoma (FTA: 72.9%) tissues, whilst in a majority of nodular goitre (69.2%) and Hashimoto's thyroiditis (HT: 73.7%) thyroid tissues, it remained unaltered. GSH increased in PTC (55.3%), remained unaltered in FTA (97.3%) and all other goiter samples studied. SOD increased in PTC (51.1%) and all other malignant thyroid tissues studied. CAT remained unaltered in PTC (95.7%), FTA (97.3%) and all other non-malignant samples (HT, MNG, TMNG) studied. GPx increased in PTC (63.8%), all other malignant thyroid tissues and remained unaltered in many of the FTA (91.9%) tissues and all other non-malignant samples (HT, MNG, TMNG) studied. CONCLUSIONS: In the case of non-malignant thyroid tumours, the oxidant-antioxidant balance was undisturbed, whilst in malignant tumours the balance was altered, and the change in r value observed in the LPO and SOD pairs between normal and PTC tissues and also in many pairs with multi-nodular goitre (MNG)/toxic MNG tissues may be used as a marker to differentiate/detect different malignant/non-malignant thyroid tumours.

Trypsin inhibitor from edible mushroom Pleurotus floridanus active against proteases of microbial origin.

Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and food preservative applications. Fungi have been recognized as sources of protease inhibitors, although there are only few such reports on mushrooms. This work reports the purification and characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor toward trypsin was with Ki of 1.043 × 10(-10) M. The inhibitor was thermostable up to 90 °C with maximal stability at 30 °C, active over a pH range of 4-10 against proteases from Aspergillus oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the possibility of utilization of protease inhibitor from P. floridanus against serine proteases.

Complicated Open Supracondylar Fracture of the Femur Managed Primarily with an Autogenous Split Fibular Graft in Conjunction with an Intramedullary Nail: A Case Report.

CASE: A thirty-two-year-old man presented with an open type-IIIA Müller type-C2 supracondylar fracture of the femur and an ipsilateral segmental fracture of the tibia. An external fixator was used for initial stabilization. After ten days, the fractured femur was stabilized with a retrograde intramedullary nail along with an autogenous split fibular graft placed on either side of the nail; intramedullary nail fixation of the tibia was also performed. At the two-year follow-up, both fractures had united. CONCLUSION: An autogenous split fibular graft in conjunction with an intramedullary nail is a viable option to manage bone defects in complicated supracondylar fractures of the femur.

Comparison of three hypothermic target temperatures for the treatment of hypoxic ischemia: mRNA level responses of eight genes in the piglet brain.

Hypothermia can reduce neurodevelopmental disabilities in asphyxiated newborn infants. However, the optimal cooling temperature for neuroprotection is not well defined. We studied the effects of transient piglet brain hypoxic ischemia (HI) on transcriptional activity of eight genes and if mRNA level alterations could be counteracted by whole body cooling to 35, 33.5 or 30 °C. BDNF mRNA was globally upregulated by the insult, and none of the cooling temperatures counteracted this change. In contrast, MANF mRNA was downregulated, and these changes were modestly counteracted in different brain regions by hypothermic treatment at 33.5 °C, while 30 °C aggravated the MANF mRNA loss. MAP2 mRNA was markedly downregulated in all brain regions except striatum, and cooling to 33.5 °C modestly counteract this downregulation in the cortex cerebri. There was a tendency for GFAP mRNA levels in core, but not mantle regions to be downregulated and for these changes to be modestly counteracted by cooling to 33.5 or 35 °C. Cooling to 30 °C caused global GFAP mRNA decrease. HSP70 mRNA tended to become upregulated by HI and to be more pronounced in cortex and CA1 of hippocampus during cooling to 33.5 °C. We conclude that HI causes alterations of mRNA levels of many genes in superficial and deep piglet brain areas. Some of these changes may be beneficial, others detrimental, and lowering body temperature partly counteracts some, but not all changes. There may be general differences between core and mantle regions, as well as between the different cooling temperatures for protection. Comparing the three studied temperatures, cooling to 33.5 °C, appears to provide the best cooling temperature compromise.

Screening of selected medicinal plants for in vitro antidermatophytic activity.

Different solvent extracts of leaves of Achyranthes aspera, Aegle marmelos, Cleistanthus collinus, Curcuma aromatica and Strychnos nux-vomica were screened against dermatophytes viz., Trichophyton mentagrophytes, T. rubrum, Microsporum gypseum, M. canis and Epidermophyton floccossum var. nigricans. The mean zones of inhibition were between 7.1 and 26.5mm. Minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) were from 7.81 to 500 and from 15.62 to 1000µg/mL respectively. The highest mean zone of inhibition (26.5mm), the lowest MIC value (7.81µg/ml) and the lowest MFC (15.62µg/ml) were observed in ethyl acetate extract of A. aspera against T. rubrum. The standard antifungal drug ketoconazole (10µg/disc) was used as the positive control and mean zones of inhibition were from 23 to 29mm. Further separation of active principle from ethyl acetate extract of A. aspera is under progress.

Valorization of date palm (Phoenix dactylifera) fruit processing by-products and wastes using bioprocess technology - Review.

The date palm Phoenix dactylifera has played an important role in the day-to-day life of the people for the last 7000 years. Today worldwide production, utilization and industrialization of dates are continuously increasing since date fruits have earned great importance in human nutrition owing to their rich content of essential nutrients. Tons of date palm fruit wastes are discarded daily by the date processing industries leading to environmental problems. Wastes such as date pits represent an average of 10% of the date fruits. Thus, there is an urgent need to find suitable applications for this waste. In spite of several studies on date palm cultivation, their utilization and scope for utilizing date fruit in therapeutic applications, very few reviews are available and they are limited to the chemistry and pharmacology of the date fruits and phytochemical composition, nutritional significance and potential health benefits of date fruit consumption. In this context, in the present review the prospects of valorization of these date fruit processing by-products and wastes' employing fermentation and enzyme processing technologies towards total utilization of this valuable commodity for the production of biofuels, biopolymers, biosurfactants, organic acids, antibiotics, industrial enzymes and other possible industrial chemicals are discussed.

Halocin SH10 production by an extreme haloarchaeon Natrinema sp. BTSH10 isolated from salt pans of South India.

Halobacteria, members of the domain Archaea that live under extremely halophilic conditions, are often considered as dependable source for deriving novel enzymes, novel genes, bioactive compounds and other industrially important molecules. Protein antibiotics have potential for application as preserving agents in food industry, leather industry and in control of infectious bacteria. Halocins are proteinaceous antibiotics synthesized and released into the environment by extreme halophiles, a universal characteristic of halophilic bacteria. Herein, we report the production of halocin (SH10) by an extremely halophilic archeon Natrinema sp. BTSH10 isolated from salt pan of Kanyakumari, Tamilnadu, India and optimization of medium for enhanced production of halocin. It was found that the optimal conditions for maximal halocin production were 42 °C, pH 8.0, and 104 h of incubation at 200 rpm with 2% (V/V) inoculum concentration in Zobell's medium containing 3 M NaCl, Galactose, beef extract, and calcium chloride as additional supplements. Results indicated scope for fermentation production of halocin for probable applications using halophilic archeon Natrinema sp. BTSH10.

Detergent compatible alkaline lipase produced by marine Bacillus smithii BTMS 11.

Bacillus smithii BTMS 11, isolated from marine sediment, produced alkaline and thermostable lipase. The enzyme was purified to homogeneity by ammonium sulfate precipitation and ion exchange chromatography which resulted in 0.51 % final yield and a 4.33 fold of purification. The purified enzyme was found to have a specific activity of 360 IU/mg protein. SDS-PAGE analyses, under non-reducing and reducing conditions, yielded a single band of 45 kDa indicating the single polypeptide nature of the enzyme and zymogram analysis using methylumbelliferyl butyrate as substrate confirmed the lipolytic activity of the protein band. The enzyme was found to have 50 °C and pH 8.0 as optimum conditions for maximal activity. However, the enzyme was active over wide range of temperatures (30-80 °C) and pH (7.0-10.0). Effect of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on lipase activity was studied to determine the novel characteristics of the enzyme. More than 90 % of the enzyme activity was observed even after 3 h of incubation in the presence of commercial detergents Surf, Sunlight, Ariel, Henko, Tide and Ujala indicating the detergent compatibility of B. smithii lipase. The enzyme was also found to be efficient in stain removal from cotton cloths. Further it was observed that the enzyme could catalyse ester synthesis between fatty acids of varying carbon chain lengths and methanol with high preference for medium to long chain fatty acids showing 70 % of esterification. Results of the study indicated scope for application of this marine bacterial lipase in various industries.

Androgen receptor expression in human thyroid cancer tissues: a potential mechanism underlying the gender bias in the incidence of thyroid cancers.

Gender bias in the incidence of thyroid cancer is well known, however, the underlying mechanism is largely unknown. The current study determines variations in the molecular characteristics of thyroid cancers between men and women. Normal and cancerous thyroid tissues were collected from a total of 125 men and women who underwent surgical thyroidectomy. Testosterone levels in serum and thyroid cancer tissues were elevated in women while it decreased in men compared to respective control groups; whereas, ligand binding activity increased in men and decreased in women. Androgen receptor (AR) mRNA expression increased in a majority of men while it decreased in a majority of women except those with follicular thyroid carcinoma (FTC). In thyroid cancers of women, Pearson's correlation analysis showed a positive correlation of AR mRNA with AR protein, CBP and Sp1, whereas AR mRNA showed a negative correlation with p53. In case of men, AR mRNA showed a positive correlation with AR and cyclin D1 proteins in papillary thyroid carcinoma (PTC); and CBP and Sp1 in follicular thyroid adenoma (FTA), whereas AR mRNA showed a positive correlation with p53. Our study identified for the first time that AR is posttranscriptionally regulated by miR-124a in thyroid cancer tissues. Further, our in vitro studies with a PTC cell line (NPA-87-1) showed miR-124a as the potent inhibitor of AR that impairs cell proliferation even in the presence of testosterone. Thus, the current study suggests that: (i) the varying pattern of testosterone level and AR status in thyroid tissues of men and women may predispose to the gender specific incidence of thyroid tumors and (ii) miR-124a plays a significant role in determining the AR gene expression pattern and thus, androgen mediated thyroid tumor growth.

In vitro anti-plasmodial activity of some traditionally used medicinal plants against Plasmodium falciparum.

The anti-plasmodial activity of different solvent extracts of Adhatoda vasica (root), Caesalpinia pulcherrima (leaf), Carica papaya (pulp), Erythroxylum monogynum (leaf), Lantana camara (whole plant), Ocimum sanctum (root) and Phyllanthus niruri (whole plant) were studied against Plasmodium falciparum. Of the 35 extracts tested, seven extracts showed good anti-plasmodial activity. Methanol extract of C. pulcherrima showed the lowest IC50 value (10.96 µg/mL) followed by methanol extract of A. vasica (IC(50)=11.1 µg/mL), chloroform extract of O. sanctum (IC(50)=11.47 µg/mL), methanol extract of E. monogynum (IC(50)=12.23 µg/mL), acetone extract of C. pulcherrima (IC(50)=12.49 µg/mL), methanol extract of O. sanctum and acetone extract of A. vasica (IC(50)=14.04 µg/mL). The results of the present study justify the use of these medicinal plants in traditional practice, and also, a further study on the isolation of anti-plasmodial molecules from their active crude extracts is in progress.

Antibacterial and antifungal efficacy of fatty acid methyl esters from the leaves of Sesuvium portulacastrum L.

OBJECTIVES: The fatty acid methyl esters (FAME extract) from Sesuvium (S.) portulacastrum was studied for its fatty acid composition and antimicrobial activity against human pathogenic microorganisms. MATERIALS AND METHODS AND RESULTS: The gas chromatographic analysis of FAME extract revealed the presence of palmitic acid with the highest relative percentage (31.18%), followed by oleic acid (21.15%), linolenic acid (14.18%) linoleic acid (10.63%), myristic acid (6.91%) and behenic acid (2.42%). The saturated fatty acids were higher than the unsaturated fatty acids. FAME extract showed the highest antibacterial and anticandidal activities and moderate antifungal activity against the tested microorganisms. The highest mean zone of inhibition (16.3 mm) and the lowest MIC (0.25 mg/ml) and MBC (0.5 mg/ml) values were recorded against Bacillus subtilis. The lowest mean zone of inhibition (8.8 mm) and the highest MIC (8 mg/ml) and MFC (16 mg/ml) values were recorded against Aspergillus fumigatus and Aspergillus niger. CONCLUSIONS: The results of the present study justify the use of S. portulacastrum in traditional medicine and the FAME extract can be used as a potential antimicrobial agent against the tested human pathogenic microorganisms.

Lipase from marine Aspergillus awamori BTMFW032: production, partial purification and application in oil effluent treatment.

Marine fungus BTMFW032, isolated from seawater and identified as Aspergillus awamori, was observed to produce an extracellular lipase, which could reduce 92% fat and oil content in the effluent laden with oil. In this study, medium for lipase production under submerged fermentation was optimized statistically employing response surface method toward maximal enzyme production. Medium with soyabean meal-0.77% (w/v); (NH(4))(2)SO(4)-0.1m; KH(2)PO(4)-0.05 m; rice bran oil-2% (v/v); CaCl(2)-0.05 m; PEG 6000-0.05% (w/v); NaCl-1% (w/v); inoculum-1% (v/v); pH 3.0; incubation temperature 35°C and incubation period-five days were identified as optimal conditions for maximal lipase production. The time course experiment under optimized condition, after statistical modeling, indicated that enzyme production commenced after 36 hours of incubation and reached a maximum after 96 hours (495.0 U/ml), whereas maximal specific activity of enzyme was recorded at 108 hours (1164.63 U/mg protein). After optimization an overall 4.6-fold increase in lipase production was achieved. Partial purification by (NH(4))(2)SO(4) precipitation and ion exchange chromatography resulted in 33.7% final yield. The lipase was noted to have a molecular mass of 90 kDa and optimal activity at pH 7 and 40°C. Results indicated the scope for potential application of this marine fungal lipase in bioremediation.

Propyl gallate synthesis using acidophilic tannase and simultaneous production of tannase and gallic acid by marine Aspergillus awamori BTMFW032.

Marine Aspergillus awamori BTMFW032, recently reported by us, produce acidophilic tannase as extracellular enzyme. Here, we report the application of this enzyme for synthesis of propyl gallate by direct transesterification of tannic acid and in tea cream solubilisation besides the simultaneous production of gallic acid along with tannase under submerged fermentation by this fungus. This acidophilic tannase enabled synthesis of propyl gallate by direct transesterification of tannic acid using propanol as organic reaction media under low water conditions. The identity of the product was confirmed with thin layer chromatography and Fourier transform infrared spectroscopy. It was noted that 699 U/ml of enzyme could give 60% solubilisation of tea cream within 1 h. Enzyme production medium was optimized adopting Box-Behnken design for simultaneous synthesis of tannase and gallic acid. Process variables including tannic acid, sodium chloride, ferrous sulphate, dipotassium hydrogen phosphate, incubation period and agitation were recognized as the critical factors that influenced tannase and gallic acid production. The model obtained predicted 4,824.61 U/ml of tannase and 136.206 µg/ml gallic acid after 48 h of incubation, whereas optimized medium supported 5,085 U/ml tannase and 372.6 µg/ml of gallic acid production after 36 and 84 h of incubation, respectively, with a 15-fold increase in both enzyme and gallic acid production. Results indicated scope for utilization of this acidophilic tannase for transesterification of tannic acid into propyl gallate, tea cream solubilisation and simultaneous production of gallic acid along with tannase.

Characterization of an extracellular alkaline serine protease from marine Engyodontium album BTMFS10.

An alkaline protease from marine Engyodontium album was characterized for its physicochemical properties towards evaluation of its suitability for potential industrial applications. Molecular mass of the enzyme by matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) analysis was calculated as 28.6 kDa. Isoelectric focusing yielded pI of 3-4. Enzyme inhibition by phenylmethylsulfonyl fluoride (PMSF) and aprotinin confirmed the serine protease nature of the enzyme. K (m), V (max), and K (cat) of the enzyme were 4.727 x 10⁻² mg/ml, 394.68 U, and 4.2175 x 10⁻² s⁻¹, respectively. Enzyme was noted to be active over a broad range of pH (6-12) and temperature (15-65 °C), with maximum activity at pH 11 and 60 °C. CaCl2 (1 mM), starch (1%), and sucrose (1%) imparted thermal stability at 65 °C. Hg²âº, Cu²âº, Fe³âº, Zn²âº, Cd⁺, and Al³âº inhibited enzyme activity, while 1 mM Co²âº enhanced enzyme activity. Reducing agents enhanced enzyme activity at lower concentrations. The enzyme showed considerable storage stability, and retained its activity in the presence of hydrocarbons, natural oils, surfactants, and most of the organic solvents tested. Results indicate that the marine protease holds potential for use in the detergent industry and for varied applications.

Protease inhibitor from Moringa oleifera with potential for use as therapeutic drug and as seafood preservative.

Protease inhibitors are well known to have several applications in medicine and biotechnology. Several plant sources are known to return potential protease inhibitors. In this study plants belonging to different families of Leguminosae, Malvaceae, Rutaceae, Graminae and Moringaceae were screened for the protease inhibitor. Among them Moringa oleifera, belonging to the family Moringaceae, recorded high level of protease inhibitor activity after ammonium sulfate fractionation. M. oleifera, which grows throughout most of the tropics and having several industrial and medicinal uses, was selected as a source of protease inhibitor since so far no reports were made on isolation of the protease inhibitor. Among the different parts of M. oleifera tested, the crude extract isolated from the mature leaves and seeds showed the highest level of inhibition against trypsin. Among the various extraction media evaluated, the crude extract prepared in phosphate buffer showed maximum recovery of the protease inhibitor. The protease inhibitor recorded high inhibitory activity toward the serine proteases thrombin, elastase, chymotrypsin and the cysteine proteases cathepsin B and papain which have more importance in pharmaceutical industry. The protease inhibitor also showed complete inhibition of activities of the commercially available proteases of Bacillus licheniformis and Aspergillus oryzae. However, inhibitory activities toward subtilisin, esperase, pronase E and proteinase K were negligible. Further, it was found that the protease inhibitor could prevent proteolysis in a commercially valuable shrimp Penaeus monodon during storage indicating the scope for its application as a seafood preservative. This is the first report on isolation of a protease inhibitor from M. oleifera.