Effect of Time and Temperature on Biodiesel Production using Melon (Cucumeropsismannii), Groundnut (Arachis hypogea) and Soya bean (Glycine max)

This study investigated the optimum condition for biodiesel production at varying temperatures and time using melon (Cucumeropsismannii), groundnut (Arachis hypogea), and soya bean (Glycine max) seed oils. Extraction of oil from Cucumeropsismannii, Arachis hypogea, and Glycine max was accomplished using n-hexane (67.7-69.2oC) as the solvent. Biodiesel was produced from the three different seed oils at varying temperatures of 65oC, 55oC, and 45oC and also at the varied time of 60mins, 50mins and 40mins. The best percentage yield was obtained at a temperature of 65oC and a period of 60 minutes. At 40 min, the process was not complete. A good number of the transesterification process was completed at 50 mins. Also, at the lower temperature of 45oC, the method was not complete. The maximum % yield of the biodiesel obtained was 90.83% for Glycine max, 78.00% for Arachis hypogea, and 77.58% for Cucumeropsismannii seed oils. Fuel properties such as kinematic viscosity, pour point, carbon residue, cloud point, water content, flash point, cetane index, and sulfated ash were examined on the biodiesel. The flash point, carbon residue, kinematic viscosity, and water content were within the standard specified for petrol diesel. Cloud point and pour points of this product were found to be greater than that of petrol diesel. The cetane index was lower than the standard specified for petrol diesel and the three samples contained no sulfated ash. Therefore, melon (Cucumeropsismannii), groundnut (Arachis hypogea), and soya bean (Glycine max) are good alternatives to biodiesel production. Copyright (c) The Authors

Recent Developments of Recombinant Protein Expression for Therapeutic Purposes

In the most recent seven to eight years, the therapeutic recombinant proteins have rapidly expanded in the biotechnology domain due to its wide variety of needs. There has been significant development in the mammalian expression system for fine purification and increased level of expressed recombinant proteins [1,2]. Many drugs like tetracycline have been demonstrated on the Chinese Hamster Ovary cell line for promising multi control strategies and effective cytotoxicity. Mammalian expression system improves the proper glycosylation of recombinant proteins which are very helpful to increase solubility of product [3-6]. Meanwhile on the prokaryotic expression system, E. coli has proven to be an easier to handle, friendly and economical strain [2]. Recently these expression systems are using to work on antibody fragment productions and their proper folding with co-expression of chaperones [7]. Moreover E. coli has been used for the production of cancer cell penetrating peptides which promises the targeted delivery of drugs to specific effector cells only. Yeast systems are also being used for the antibody fragments production and the high level production of insulin. Interestingly cell free expression systems are also participating in this game and that would be very fascinating to see in the coming years about cell extract medium for production of high level recombinant protein [8, 9]. Purification and optimization of recombinant protein has always been a challenging situation for scientists and they paid more attention to increase the overall yield of the product. Many affinity chromatography techniques has been introduced for efficient purification of protein of interest [10]. Despite these research and developments in methodologies to produce and purify the recombinant therapeutic protein, scientists still face the hurdles and challenges with all expression systems. Rationally E. coli produces inclusion bodies and many mammalian cell types do not show the same results with the same recombinant protein. [11]. So there is a requirement for adding the appropriate features to the expression systems focused to better improvising recovery, production and purification of recombinant protein. Copyright(c) The Author

COVID-19: Immune Perspective and Current Treatment Strategies

Coronavirus disease 2019 (COVID-19) is now a well-established and globally lethal respiratory pandemic, affecting millions with about 10% fatality rate. Infected patients show mild to severe symptoms that may manifest as mild fever, cough, headache and nausea or they may even remain asymptomatic. World Health Organization has reported over 245,373,039 confirmed cases worldwide with 4,979,421 deaths (October 2021). There are myriads of promising approaches to pharmacologically treat the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the newly reported seventh human coronavirus, which is responsible for this pandemic. Various types of potential drugs; monoclonal antibodies, interferon therapies, peptides, small molecule drugs, oligonucleotides and vaccines are under consideration and target various structural components of the virus. A strong host immune system is a key player in combating COVID-19 along with the effective vaccines that are a game-changing tool. The focus of the review is some worth mentioning drug candidates; remdesivir, lopinavir, emetine, aloxistatin, viracept, homoharringtonine, ivermectin, favipiravir, tocilizumab, chloroquine and leronlimab against COVID-19 infections, targeting the membrane nucleocapsid, spike or envelope proteins, either currently in clinical trials or under consideration. Many drugs directly inhibit the viral infection while others trigger the immune system to fight against the virus. Furthermore, we also discussed the current covid vaccines; Pfizer, Moderna, etc. from different technical lines for immunization. Therefore, here we review how the immune system tries to manage the infection as well as vaccines and some of the potential therapeutic agents. Copyright(c) The Authors

Cloning, Amplified Expression and Bioinformatics Analysis of a Putative Nucleobase Cation Symporter-1 (NCS-1) Protein From Rhodococcus erythropolis

The Rhodococcus erythropolis gene DYC18_RS18060 (1437 bp) putatively codes for a secondary transporter of the Nucleobase Cation Symporter-1 (NCS-1) protein family (478 amino acids). The DYC18_RS18060 gene was successfully cloned from R. erythropolis genomic DNA with addition of EcoRI and PstI restriction sites at the 5′ and 3′ ends, respectively, using PCR technology. The amplified gene was introduced into IPTG-inducible plasmid pTTQ18 immediately upstream of the sequence coding for a His6-tag. The construct was transformed into Escherichia coli BL21(DE3), then amplified expression of the DYC18_RS18060-His6 protein was achieved with detection by SDS-PAGE and western blotting. Computational methods predicted that DYC18_RS18060 has a molecular weight of 51.1 kDa and isoelectric point of 6.58. The protein was predicted to be hydrophobic in nature (aliphatic index 113.24, grand average of hydropathicity 0.728) and to form twelve transmembrane spanning α-helices with both N- and C-terminal ends at the cytoplasmic side of the membrane. Whilst database sequence similarity searches and phylogenetic analysis suggested that the substrate of DYC18_RS18060 could be cytosine, this was not certain based on comparisons of residues involved in substrate binding in experimentally characterised NCS-1 proteins. This study has laid foundations for further structural and functional studies of DYC18_RS18060 and other NCS-1 proteins. Copyright(c) The Authors

Implementation of Response Surface Methodology for Enhanced Production of Endoglucanase by Thermophilic Aspergillus Fumigatus

Enzymes are biocatalysts which play key roles in the body of living organisms. Cellulose is major source of plant biomass. Its β-1,4-glucosidic bonds are hydrolyzed by cellulases. These cellulases can be produced by a variety of microorganisms including fungi, bacteria and actinomycetes which are used today for the industrial applications. The current study was aimed to optimize the cultural conditions for maximum production of endoglucanase by Aspergillus fumigatus through solid state fermentation of sugarcane bagasse. Response Surface Methodology (RSM) was employed under Central Composite Design (CCD) for the optimization of growth including pH, temperature, time period & inoculum size and nutritional parameters including glucose, fructose and (NH₄)₂SO₄. The effect of different metal ions on endoglucanase production was also monitored. It was partially purified by (NH₄)₂SO₄ precipitation and gel filtration chromatography. Finally, edogucanase was characterized for optimum pH, temperature and determination of kinetic parameters. Maximum enzyme activity was found as 0.9 IU/mL/min in the presence of 6 g substrate, 3.5 mL inoculum, 4.5 pH, 40 °C temperature at incubation time of 84 hrs. After addition of carbon and nitrogen sources enzyme activity increased to 1.4 IU/mL/min. It was further increased to 1.56 IU/mL/min with 0.42% of CaCl2. Maximum purification was achieved at 50% saturation by ammonium sulphate (NH₄)₂SO₄. Optimum temperature and pH were 40 °C and 5 respectively, whereas the values for Km and Vmax were 5.37 mM and 696 uM/mL/min., respectively. These findings suggested that endoglucanase by Aspergillus fumigatus could be suitable for various industries. Copyright (c) The Authors

The Production and characterization of Polyclonal Antibodies Against Interferon Alpha in Mice

The polyclonal antibodies are used extensively for research purposes in many areas of biology, such as immunoprecipitation, histochemistry, enzyme linked immunosorbent assays (ELISA), diagnosis of disease and western blots. Typically, an animal’s immune system will generate a large group of antibodies that recognize several epitopes of a particular antigen. Interferon alpha plays an important role in immune response activation and therefore is of interest in studies related to autoimmune diseases. In this paper the production of antibodies against interferon was studied in order to quantify interferon production to analyze interferon levels in autoimmune disorders in the future. For the antibody production, one month old laboratory grade mice were injected with interferon alpha in combination with a Freund’s complete adjuvant for a course of five weeks after which the antibodies were obtained in mouse serum. Confirmation of the production of anti-interferon alpha antibodies was carried out by the Elisa, immune dot blot and western blot analysis. An interferon alpha of approximately 20.5-21.5 KDa was detected in immunedot blot test. These antibodies may be produced in these mouse models commercially and could be used in future for treatment of autoimmune diseases by managing the interferon levels in the patients. Copyright(c) The Authors

Prevalence of ABO and Rh blood groups and their association with Transfusion-Transmissible Infections (TTIs) among Blood Donors in Islamabad, Pakistan

Certain Rh positive blood groups showed a link between the ABO blood grouping and susceptibility to some infectious ailments. Study was carried out to find the association of ABO Blood Group System / Rh type with different viral infection. A retrospective observational study was carried in 7631 blood donors to find the association of ABO Blood Group System / Rh type with viral infection due to hepatitis B virus, hepatitis C virus, human immunodeficiency virus and Syphilis. Data of the study indicateF that the highest incidence of blood group in blood donor was B followed by O, A and AB. The incidence of HBsAg +ve, HCV Ab +ve and infection of Syphilis was also highest in donor with blood group B followed by O, A and AB. Blood group A were more susceptible to HIV Ab +ve followed by blood group B, O and AB. The group B donors had the greatest risk of viral infection with HBV, HCV, HIV, and Syphilis; followed by the groups O, A, and AB. Donors in group AB had the lowest incidence rate of all viral infection. The study revealed that blood group B is most prevalent among donors, followed by blood groups O, A, and AB. On the other side, the rate of HBV, HCV, HIV, and Syphilis infection was greatest among group B donors, followed by group O, group A, and group AB. Copyright(c) The Authors

Efficacy of Ethanolic plant extracts of Zingiber officinale, Raphanus sativus, Rosa indica and Aloe vera against Heterotermes indicola

Present study was performed to evaluate the anti-termitic potential of four common plant species of Zingiber officinale, Raphanus sativus, Rosa indica and Aloe vera. During laboratory bioassay, the biological activity of ethanolic extracts of these plants was evaluated. The ethanol extract of Z. officinale caused highest mortality (100%) while least mortality (68%) was observed in R. indica extract. The LT50 for Z. officinale were 24.34, 40.40 and 64.7 h for concentrations of 10%, 5% and 3% respectively. Present findings suggested that these plant extracts can provide environmental friendly management of H. indicola. In A. vera, also resulted in significant mortality against H. indicola i.e. 94%, 64% and 44% on exposure of 10%, 5% and 3% concentraions. While LT50 were 34.66, 65.59, and 100.1 hours against 10%, 5% and 3% concentrations. In R. sativus, mortality of H. indicola was 76%, 50% and 30% while LT50 against were 59.10, 86.20, and 127.3 h, against concentration of 10%, 5% and 3%. Least mortality was observed in R. indica i.e. 68%, 44% and 30% while LT50 were 64.17, 94.58 and 125.5 h at 10%, 5% and 3% respectively. This study revealed that the extracts of Z. officinale, A.vera and R. Sativus have a potential to be used for termite control especially Heterotermes indicola to minimize the damage. Copyright(c) The Author

Knowledge, Attitude and Practices of University students towards Antibiotic Use and Resistance: Questionnaire Based Assessment

Antibiotic resistance is a serious health crisis all over the world and students of biological sciences should have proper awareness to combat the problem. This study aims to assess the knowledge, attitude and practices (KAP) towards antibiotic use and resistance among university students of biological sciences in Pakistan. A descriptive cross-sectional study was conducted amongst 826 students of biological sciences programs from eight major Pakistani universities during September-November 2020. A self-administered online questionnaire was developed which consisted of four dimensions: sociodemographic characteristics (3 questions) and assessment of the knowledge, attitude and practices with 13, 6 and 11 questions, respectively. Association of KAP scores with sociodemographic characteristics was analyzed by nonparametric analysis i.e., Mann-Whitney U and Kruskal-Wallis H test (p-value < 0.05). The sample comprised more females (82.9%) than males while most of the respondents belonged to 21-23 years and enrolled in the master’s degree. The overall knowledge score (correct answer) was 74.7% which was significantly associated with gender, age and degree. The positive attitude score (87.9%) was significantly higher in >23 years age group. Around 75% of the surveyed students had positive practices. The scores were observed significantly higher in females, 21-23 years age group and doctoral students. The study provides baseline evidence about the knowledge, attitudes and practices regarding antibiotic use and resistance among biology students. Overall, good positive attitude was depicted by participants however level of knowledge and positive practices was found lower and declared as moderate. Academic interventions such as lectures, courses, seminars and workshops on antibiotic use, along with establishing open-access antibiotic-resistance learning sites and use of mainstream and social media platforms, etc., are needed to improve the awareness and practices of university students of biological sciences with regards to the rational use of antibiotics.

Isolation, Identification and Characterization of Urinary Tract Infectious Bacteria and the Effect of Different Antibiotics

Etiological patterns of uropathogens is different in different regions due to continuous evolution, of bacteria, antibiotics sensitivity patterns, misuse and overuse of antibiotics. Therefore, it is important to know the antibiotic susceptibility patterns for prescription of suitable antibiotic. This study was conducted to determine the prevalence of uropathogens and their antimicrobial sensitivity pattern from Kohat region of Pakistan. In this study 100 samples were collected from both males and females of all ages in which 70 samples contained microbes. In 30 samples no microbial growth was recorded. The Percentage of positive culture from both male and female were 57% and 43% respectively Both Gram (+) and Gram (-) bacteria were found in UTI but E.coli (34.21%) was predominant followed by K. pneumoniae (10.52%), P. aeruginosa (9.21%), K. oxytoca (6.57%), C. albicans (5.26%), E. faecium (5.26%), E. faecalis (3.94%), S. aureus (3.94%), E. cloacea (2.63%), C. freundii (2.63%), P. mirabalis (2.63%) and A. baumannii (1.31%). Many of the isolates showed resistance to commonly used antibiotics. The sensitivity percentage of different commonly used antibiotics against both Gram (+) and Gram(-) bacteria were Ampicillin 13%, Ceftriaxone 25%, Amikacin 77%, Gentamicin 41%, Augmentin 44.77%, Fosfomycin 64%, Cotrimoxazole 36%, Nitrofurantoin 68%, Ciprofloxacin 37%, Imipenem 78%, Meropenem 67%, Cefepime 25% and Tetracycline 40%. The most effective antibiotics against both Gram (+) and Gram (-) bacteria were Fosfomycin, Imipenem, Meropenem Amikacin and Nitrofurantoin. In light of the findings of this study, it is strongly recommended to discover new antimicrobial compounds and evaluate the resistant pattern at genomic and proteomics level to discover the genes which are responsible for antibiotics resistant pattern.