Green and efficient synthesis of cellulose nanocrystals from Hamelia patens leftover via hydrolysis of microwave assisted-ionic liquid (MWAIL) pretreated microcrystalline cellulose.

The efficient bioconversion of the lignocellulosic agro-waste has immense importance in biorefinery processing in extracting the cellulose and saccharide fractions. To achieve this, a series of chemical pretreatments is employed, thus concerning environmental threats limit its use. Therefore, an ionic liquid is employed for pretreatment before sustainable extractions owing to its safe manipulation, recycling, and reusability. Specifically, microwave-assisted ionic liquid (MWAIL) pretreatment has significant importance in extracting high cellulose yield at less thermal power consumption. In this study, the leftover stalks of Hamelia patens were subjected to MWAIL pretreatment at 60, 70, 80, and 90 °C to extract microcrystalline cellulose (MCC). Subsequently, the MCC was fabricated into cellulose nanocrystals (CNC) through hydrolytic treatment using acidic and ionic liquids and denoted as CNC-AH and CNC-ILH. Thus obtained CNC was characterized by FTIR, FESEM, XRD, and TGA to investigate the influence of solvent on its morphology, crystallinity, and thermal stability of CNC. The results support that the CNC-ILH has comparatively more thermal and dispersal stability with a reduced crystallinity index than CNC-AH. The surprising results of CNC-ILH signify its utilization in diverse applications in the food and industrial sectors.

Sono-Microwave Assisted Chlorine free and Ionic Liquid (SMACIL) extraction of cellulose from Urtica dioica: A benign to green approach.

The extraction of cellulose using eco-friendly solvents has been gaining significant attention for a couple of decades. This study investigated the impact of benign and green solvents on the extraction, thermal stability, mechanical properties, and crystallinity of cellulose extracted from Urtica dioica (Stinging nettle) using a Sono-Microwave Assisted Chlorine free and Ionic Liquid (SMACIL) extraction technique. In this regard, the stalks were undergone through pre chemical treatment before starting bleaching them with hydrogen peroxide (HPO) and 1-butyl-3-methylimidazolium acetate (BMIM-Ac) having different mole ratios (5, 7.5, and 10) to expose cellulose. The Urtica dioica cellulose (UDC) was characterized using FTIR, tensile testing, FESEM, XRD, and TGA. The fibrillation and lumen can be seen in SEM images that confirm the extraction of cellulose. The results showed that the BMIM-Ac-10 gives the maximum cellulose yield (88 %) than other compositions. Moreover, the cellulose extracted using BMIM-Ac-10 has high mechanical strength which makes it a potential constituent for various applications in the field of materials science. These results have significant implications for the development of sustainable and efficient processes for the extraction of cellulose.

Purification and Characterization of Keratinase from Bacillus licheniformis dcs1 for Poultry Waste Processing.

Feather wastes-byproduct of commercial poultry processing plant is produced in large amounts. Keratinolytic enzymes produced by feather degrading bacteria can easily degrade these waste products releasing pure keratin as a residue. The aim of present study was to isolate, and characterize feather degrading bacteria as well as assess the keratinolytic potential of purified enzyme. Three feather degrading bacteria (dps3, wps1 and dcs1) were isolated from feathers of domestic chickens. Preliminary characterization of isolated bacteria revealed these isolates belonging to genus Bacillus. 16S rRNA gene sequencing identified the isolates as B. subtilis dps3 (MW255302), B. cereus wps1 (MW255303) and B. licheniformis dcs1 (MW255304). Cell free supernatant of B. licheniformis dcs1 degraded feathers completely in 14 days indicating its keratinolytic ability. Purification of keratinase enzyme from B. licheniformis dcs1 was performed using column chromatography. SDS-PAGE indicated its molecular weight as 32 KDa. Kerotinolytice activity was maximum at optimum pH of 7 and 45℃ temperature. Enzyme showed the potential to degrade keratin material such as hairs and nails of humans. Findings of current study suggested that purified enzyme possess potential to upgrade nutritional quality of poultry waste containing keratin and might play as important biotechnological tool for keratin hydrolysis.

Discrimination of Pakistani Fountain Pen Inks by Gas Chromatography-Mass Spectrometry (GC-MS).

In developing countries, the chances of fraud in written documents are comparatively high. Therefore, comparison of fountain pen inks is especially imperative in examination of forensic questioned documents. We have investigated the use of the gas chromatography-mass spectrometry technique in profiling and discrimination of fountain pen ink used in Pakistan for forensic purpose. The main purpose of this study was to discriminate different Pakistani fountain pen inks. The datum for Pakistani inks of fountain pen is not obtainable. In this research study, blue, black, and green colors fountain pen inks commercially used in Pakistan have been extracted from paper using micropunch and then investigated using the gas chromatography-mass spectrometry technique. Gas chromatography-mass spectrometry (GC-MS) was used to differentiate various brands of different colors of fountain pen inks based on their chemical composition. Molecular ion peaks for different components were obtained, and components were identified on the basis of detected ions. Results have been calculated and compared in terms of discriminating power (D.P.). The D.P. for blue, black, and green inks of fountain pen was 1.0 by using the gas chromatography-mass spectrometry technique.

DNA Barcoding: Molecular Identification and Phylogenetic Analysis of Pheretimoid Earthworm (Metaphire sp. and Amynthas sp.) Based on Mitochondrial Partial COI Gene from Sialkot, Pakistan.

The extremely difficult and challenging process is identifying pheretimoid species, genus Metaphire and Amynthas involving increased homoplasy in various morphological characteristics. The molecular identification, phylogenetic relationships, and evolutionary divergence time of earthworms belonging to the pheretimoid complex were investigated in this study using partial mitochondrial COI (cytochrome C oxidase subunit I) gene sequences ranging from 550-680 bp. Results revealed that 86 pheretimoid earthworms were morphologically different from a total of 342 mature worms. Moreover, 11 pheretimoid species were molecularly identified, including Metaphire posthuma (02), M. anomala (01), M. houlleti (02), M. californica (01), M. birmanica (02), Amynthas minimus (01), A. morrisi (01), and M. bununa (01). A phylogenetic tree was constructed with bootstrap values of 95%, which supported a monophyletic lineage of two well-supported clades formed by 12 partial COI sequences and 48 GenBank sequences using Hirudo medicinalis as an outgroup. The monophyly of these obtained genera indicated overall similarity at species level. Today, species like Amynthas, Metaphire and Pheretima have worm diversity in the form of pheretimoid earthworms, which dates to the Late Miocene (11.2-5.3 Mya) and the Pliocene (5.3-2.4 Mya). Compared to all relevant pheretimoid species, genetic p-distance values ranged from 0.0% to 0.57% (less than 1%). These low range values demonstrated that both genera Metaphire and Amynthas, supported the theory, which states that there are shared similarities among the species, despite different morphology. The current study is the first attempt in Pakistan to identify earthworms through DNA barcoding thus providing a genomic stamp. The work explored the significance of COI gene sequences to construct molecular tools that will be useful to overcome the different obstacles in morphologically similar earthworm identification and their phylogenetic study.

Cellulose extraction of Alstonia scholaris: A comparative study on efficiency of different bleaching reagents for its isolation and characterization.

The incredible benefits of Alstonia scholaris are piquing researchers' attention in extracting its cellulose and utilizing it in further therapeutic applications. This study is based on cellulose extraction from its stalks and processed through chemical pre-treatments to manifest its cellulose content by using different bleaching reagents. A comparison was made on efficiencies of three reagents and it is found that the hydrogen peroxide exposed maximum cellulose than sodium hypochlorite and sodium chlorite. The experimental results revealed that A. scholaris possess 68-70% cellulose content. FTIR spectrum shows that OH- and CH- vibrations of cellulose appeared at 3320 cm-1 & 2892 cm-1 respectively whereas SEM images show fibrillation, rough surface, and lumens in bleached fiber that attributes to the removal of lignin and hemicelluloses and confirms cellulose extraction. The XRD pattern certifies the crystalline nature and compactness of cellulose whereas tensile properties and TGA help in understanding its flexibility, mechanical strength, and thermal stability at 370 °C respectively.

Development of an Ultrasonic-Assisted Extraction Technique for the Extraction of Natural Coloring Substance Chlorophyll from Leaves of Carica papaya.

Beautiful green leaves of Papaya are the rich source of Chlorophyll. Green color of chlorophyll has been used for a very long time as a natural colorant. Carica papaya has been considered as a good example and reasonable source of natural phytochemicals, which makes it suitable to color the food items and beverages. The aim of the present investigation is to develop the process of ultrasonic extraction in combination with solid phase extraction (SPE) to extract out chlorophyll with high yield as well as high degree of clarity. Newly customized ultrasonic-assisted extraction technique for the extraction of chlorophyll from Carica papaya leaves is optimized by taking different parameters like time, temperature, solvents concentrations, and raw material under consideration. Furthermore, the extract was purified by means of SPE and examined by using UV-Vis spectrophotometer. The highest yield of chlorophyll (dye) extract was found as 40% in solvent solution having 80 mL of ethanol and 20 mL of water with 5 minutes of extraction time, 35°C of temperature, and 1 grams of raw material in the sonication bath. Furthermore, the SPE purified sample was characterized by means of the UV-Vis spectrophotometer and here the total chlorophyll content was 34 mg/g, including chlorophyll a with a concentration of 14.1246 mg/g and chlorophyll b with concentration of 19.845 mg/g respectively. Consequently, sonication method can be suggested as a good method to get better concentration of chlorophyll.

Low temperature green extraction of Acer platanoides cellulose using nitrogen protected microwave assisted extraction (NPMAE) technique.

Practicability of advanced and innovative techniques facilitates the high yield of cellulose extraction within a short period. The research aimed at the extraction of naturally abundant cellulose from Acer platanoides by "Nitrogen protected microwave assisted extraction (NPMAE)". The NPMAE uses microwaves for heating the sample and helps in fast extraction of cellulose in the presence of nitrogen atmosphere. Cellulose extraction was intensified by bleaching treatment in closed multimode NPMAE system at 100 W and 120 °C for 15 min. Experiment's result found that Acer platanoides fiber contains 70% cellulose content and diffferent analysis were studied for all chemically pre-treated fibers and found variations in results after each chemical treatment. The SEM results of bleached fibers show the rough surface due to the removal of lignin and hemicellulose. XRD pattern and FTIR analysis are in the favor of cellulose extraction and results show the presence of type I cellulose with 65% crystallinity index whileTGA and dTGA results explain that cellulose of Acer platanoides bleached fibers (APBF) is more thermally stable below 370 °C than other pre-treated fibers.

Role of modified diet and gut microbiota in metabolic endotoxemia in mice.

This study was aimed at investigating the effect of cultured gut microbiota (GM) from obese humans coupled HFD in inducing metabolic endotoxemia in humanized mice. In total, 30 strains were isolated from 10 stool samples of obese patients. Following morphological and biochemical characterization, 16S rRNA gene sequencing of six abundant isolates identified these Klebsiella aerogenes, Levilactobacillus brevis, Escherichia coli, Staphylococcus aureus, Bacillus cereus and Bacillus subtilis (MZ052089-MZ052094). In vivo trial using above isolates, known as human gut microbiota (HGM), was performed for six months. Sixteen mice were distributed into four groups, i.e., G1 (control) mice fed with chow diet, group 2 (G2) with HFD, group 3 (G3) with HFD + HGM and group 4 (G4) with chow diet + HGM. Body mass index (BMI) and plasma endotoxins were measured pre- and post-experiment. In vivo study revealed that HFD + HGM caused significant increase (3.9 g/cm at 20 weeks) in the body weight and BMI (0.4 g/cm post-experiment) of G3 mice compared to the other groups. One-way ANOVA showed significantly higher level of endotoxins (2.41, 4.08 and 3.7 mmol/L) in mice groups G2, G3 and G4, respectively, indicating onset of metabolic endotoxemia. Cecal contents of experimental mice groups showed a shift in microbial diversity as observed by all isolates belonging to either Firmicutes or Bacteroidetes phyla, respectively. In conclusion, current study reported that minor alteration in GM composition through HFD feeding and cultured GM transfer has significant impact in development of metabolic endotoxemia, possibly via modified intestinal permeability.

Formulation and Characterization of Calcium-Fortified Jelly and Its Proximate Composition and Sensory Analysis.

Calcium is a dynamic mineral. Recent discoveries designate that low intake of calcium generates deficiencies and path to other diseases. Food fortification could play a key role to overcome this problem. To cope with this deficiency problem, jellies were formulated with food-grade calcium salts and chicken eggshell powder. In the present study, three different concentrations of calcium salts, as well as eggshell powder were used to formulate jellies. The results of the sensory evaluation indicated that the two jelly products (A&D) in the current study were suitable for consumers. Results of Atomic Absorption Spectrophotometer revealed Jelly A and jelly D had 151±0.05 ppm and 133±0.06 ppm calcium concentration, respectively. Proximate analysis of Jelly A showed that it has 6.0±0.01% ash, 9.2±0.1% moisture, 0.4±0.01 g crude protein, 82.79±0.001 g crude fiber, and 0.61±0.001 g crude fat, while the jelly D that was made with chicken eggshell powder exhibited 6.0±0.01% ash, 10.1±0.1% moisture, 0.5±0.01 g protein, 84.54±0.01 g crude fiber and 1.61±0.01 g crude fat. Therefore, these two jelly A & D were greatly appreciated among other attributes. In spite of naturally available calcium-rich sources, calcium-fortified jellies can be consumed by individuals who are incapable to take sufficient calcium from their diet.

Efficacy of spent black tea for the removal of nitrobenzene from aqueous media.

Nitrobenzene (NB) is a kind of persistent organic pollutant. A ubiquitous and cost-effective substance spent black tea (SBT) was investigated for the removal of nitrobenzene from aqueous media. The maximum uptake potential of dried biomass (SBT) for NB was found to be 14.86 mg per gram (qmax) in a batch experimental set-up. Equilibration time for NB sorption was about 50 min, and optimal removal efficiency was achieved at a dosage of 2 g/L with an initial concentration of 100 mg/L of NB. Findings revealed that NB uptake increased with an increase in the temperature from 273 K to 353 K. Sorption was also found to be pH sensitive, sorption improved as the pH value changes from alkaline to acidic (from 10 to 2). Different isotherm (Langmuir, Freundlich, Temkin and Dubinin Radushkevich) and kinetic models (pseudo-1st order, pseudo-2nd order and Elovich models) were applied to experimental results; the sorption mechanism was well described by the Freundlich and pseudo-2nd order models. Moreover, Scanning electron micrographs, ATR-FTIR spectra and the results of elemental analysis also supported the efficacy of SBT as an efficient bio-sorbent for the elimination of NB from water.

Report: Central nervous system (CNS) toxicity caused by metal poisoning: Brain as a target organ.

People relate the neural disorders with either inheritance or psychological violence but there might be some other reasons responsible for the ailment of people that do not have such a background. The present study explains the chronic effect of heavy toxic metals on nervous system. During experimentation, rabbits used as laboratory animals, were given test metals in their diet. Concentration of metals given to them in the diet was less than their tolerable dietary intake. Behavioral changes were observed during experimentation. Periodic increase in the metal concentration was seen in the blood sample of rabbits. They were slaughtered after a period of eight months of slow poisoning. Histological examination of brain tissues was performed. The brain samples were analyzed by Atomic absorption spectroscopy and Inductively Coupled Plasma Mass Spectrometry to find the retention of heavy metals in mammalian brain. Concentration of lead, mercury and cadmium in the blood samples of occupationally exposed people and patients with neurological disorders at the time of neurosurgery was determined by using the same techniques. During circulation, toxic metals passes through the nerve capillaries to settle down in the brain. Heavy metals cross the blood brain barrier and 'may retain themselves in it. Brain tumors and biopsy samples of patients with neurological disorder were also analyzed to relate neurotoxicity and heavy metal poisoning. Results obtained shows that lead, mercury and cadmium retain themselves in the brain for longer period of time and are one of the causes of neurotoxicity.