Serodiagnosis of Hepatitis B Virus Using PreS Antigen from Pakistani Isolate SBS001

Background: An estimated 325 million people worldwide live with hepatitis B and/or C and approximately, 5 million people are affected with hepatitis B. in Pakistan.This study aimed at developing PreS protein from Hepatitis B Virus Pakistani isolate (SBS001) with enhanced sensitivity to detect antibodies in serum as a diagnostic method. Methods: Gene encoding PreS region from hepatitis B Virus was cloned and expressed in Escherchia coli. The recombinant protein preS-His was purified by Ni-IDA affinity chromatography. Antibodies were raised in rabbit. This protein was screened for detection of antibodies in HBV patients’ sera through ELISA. This ELISA procedure was compared with commercially available Kit used for diagnosis of HBV infection. Results: Single band purified recombinant PreS protein was obtained with high titer of antibodies raised in rabbits. This recombinant protein was used in ELISA as antigen coated on the plate. That efficiently detected antibodies present in HBV patients. It was concluded that preS-His antigen/ protein A HRP-conjugate ELISA method was more sensitive than the commercial kit for detecting the antibodies present in HBV patient sera. Conclusion: It was concluded that SBS001 PreS recombinant protein can be used in ELISA kits for detection of HBV in Pakistani population.

Cemaran Bakteri dan Residu Antibiotika Daging Babi di Pasar Tradisional Kecamatan Abiansemal dan Kuta Kabupaten Badung

Penelitian ini bertujuan untuk mengetahui tingkat cemaran bakteri dan residu antibiotik daging babi, serta memberi gambaran penanganan daging yang dijual di pasar tradisional di Kecamatan Abiansemal dan Kuta, Badung. Penelitian ini merupakan studi cross-sectional menggunakan kuisioner dan pengujian cemaran bakteri dan residu antibiotik daging babi. Data kuisioner dan sampel daging dikumpulkan dari 26 penjual dari 8 pasar tradisional, selanjutnya data dianalisis secara deskriptif. Uji cemaran bakteri dilakukan dengan menghitung nilai Total Plate Count, Coliform, Escherichia coli, Salmonella sp, Staphylococcus aureus, dan Campylobacter sp. Uji residu antibiotik dilakukan dengan metode tapis secara bioassay terhadap antibiotik tetrasiklin dan penisilin. Hasil menunjukkan nilai rerata Total Plate Count 3,7 x 105 dengan kisaran 7,3 x 103 - 1,5 x 106 koloni/gram. Rerata Coliform dan Escherchia coli adalah 1,3 x 105 dan 2,1 x 104 dengan kisaran 1,1 x 103 - 4,4 x 105 koloni/gram dan 4,5 x 101 - 9,7 x 104 koloni/gram. Cemaran Staphylococus aureus adalah negatif hingga 2 x 101 koloni/gram. Cemaran Salmonella sp dan Campylobacter sp. adalah negatif. Residu antibiotik tetrasiklin dan penisilin masing-masing sebesar 15,38% dan 11,54% dari seluruh sampel daging babi. Tingkat pemahaman dan kesadaran masyarakat terhadap penanganan dan sanitasi daging adalah 50% responden kategori cukup dan 50% kategori kurang paham dan kurang sadar tentang sanitasi. Faktor yang mempengaruhi tingkat cemaran bakteri yaitu tindakan praktik higiene dan sanitasi penjual daging babi yaitu 7,7% kategori baik, 80,8% kategori cukup dan 11,5% dengan kategori kurang. Disimpulkan bahwa daging babi di pasar tradisional Kecamatan Abiansemal dan Kuta Kabupaten Badung tercemar bakteri dan mengandung residu antibiotik, serta dilihat dari aspek kesehatan masyarakat veteriner tidak layak untuk dikonsumsi.

Novel modification by L/F-tRNA-protein transferase (LFTR) generates a Leu/N-degron ligand in Escherichia coli.

The N-degron pathways are a set of proteolytic systems that relate the half-life of a protein to its N-terminal (Nt) residue. In Escherchia coli the principal N-degron pathway is known as the Leu/N-degron pathway of which an Nt Leu is a key feature of the degron. Although the physiological role of the Leu/N-degron pathway is currently unclear, many of the components of the pathway are well defined. Proteins degraded by this pathway contain an Nt degradation signal (N-degron) composed of an Nt primary destabilizing (Nd1) residue (Leu, Phe, Trp or Tyr) and an unstructured region which generally contains a hydrophobic element. Most N-degrons are generated from a pro-N-degron, either by endoproteolytic cleavage, or by enzymatic attachment of a Nd1 residue (Leu or Phe) to the N-terminus of a protein (or protein fragment) by the enzyme Leu/Phe tRNA protein transferase (LFTR) in a non-ribosomal manner. Regardless of the mode of generation, all Leu/N-degrons are recognized by ClpS and delivered to the ClpAP protease for degradation. To date, only two physiological Leu/N-degron bearing substrates have been verified, one of which (PATase) is modified by LFTR. In this study, we have examined the substrate proteome of LFTR during stationary phase. From this analysis, we have identified several additional physiological Leu/N-degron ligands, including AldB, which is modified by a previously undescribed activity of LFTR. Importantly, the novel specificity of LFTR was confirmed in vitro, using a range of model proteins. Our data shows that processing of the Nt-Met of AldB generates a novel substrate for LFTR. Importantly, the LFTR-dependent modification of T2-AldB is essential for its turnover by ClpAPS, in vitro. To further examine the acceptor specificity of LFTR, we performed a systematic analysis using a series of peptide arrays. These data reveal that the identity of the second residue modulates substrate conjugation with positively charged residues being favored and negatively charged and aromatic residues being disfavored. Collectively, these findings extend our understanding of LFTR specificity and the Leu/N-degron pathway in E. coli.

GC-MS Analysis and Antimicrobial Activities of Ethanol Alkaloid Leaf Extracts of Delonix elata.L

Delonix elata L., belongs to family Fabaceae used by the traditional various medicinal practices to cure jaundice, skin disease, heart disease, cancer cell formation, physiological abnormalities, heptoprotective, bronchial and rheumatic problems. The present study was screen the antimicrobial and phytochemical activity of alkaloid leaf extracts. This extracts was assessed on multidrug resistant clinical isolated from both gram positive, gram negative and antifungal strains including Bacillus subtilis, Staphylococcus aureus, Escherchia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. The zone of inhibition was determined by Agar well diffusion method with various concentration. GC- MS analysis was performed to identify major bioactive compounds present in the extracts. The GC – MS studies shown the present of 25 compound were identified in the leaf extract composition. The antimicrobial analysis revealed that C. albicans showed a highest zone of inhibition 25mm at 100 mg/ml of extracts. Present finding suggest that D. elata as plant pharmaceutical and pharmacological importance.

Green synthesis of MgO nanoparticles using Moringa oleifera leaf aqueous extract for antibacterial activity

Nanoparticle fabrication using plant extracts is an important alternative method because it is non-toxic, biocompatible, and environmentally friendly. In this study, green synthesis of MgO nanoparticles using Moringa oleifera leaf water extracts was conducted by mixing the extract and a solution of magnesium chloride. The product was characterized using different techniques, i.e. UV-Visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The UV-Vis spectrum of MgO nanoparticles shows an absorption at 280 nm. The size of the synthesized MgO nanoparticles ranges from 20-50 nm. The antibacterial activity of MgO nanoparticles was seen from the zone of inhibition against Staphylococcus aureus (6.3 mm) and against Escherichia coli (6 mm). MgO nanoparticles have been successfully fabricated using Moringa oleifera leaf aqueous extracts, providing an alternative method for synthesizing MgO nanoparticles. KEY WORDS: Antibacterial activity, Escherchia coli, MgO nanoparticles, Moringa oleifera, Staphylococcus aureus Bull. Chem. Soc. Ethiop. 2021, 35(1), 161-170. DOI: https://dx.doi.org/10.4314/bcse.v35i1.14

Bacteria Identification In Traditional Sauce (Sambal) At The Restaurants Nearby The Traditional Markei Of Gorontalo City

The unsafe food will cause the health problems in the community. Sambal which is known as the traditional sauce is a sauce made from chilies that are crushed until the water content comes out, giving it a spicy taste. Furthermore, Sambal is processed in a simple way so that it can leac to the contamination of microorganisms such as Coliform bacteria contamination. This study aims to determine the presence of Coliform bacteria contamination in Sambal at the supermarket nearby the traditional market in Gorontalo City. This research is a descriptive study using a qualitative approach. It examines the existence of Coliform bacteria using the Most Probable Number (MPN) method. The population in this study were 14 samples with the sampling technique by accidental sampling. Coliform bacteria examination results show that Sambal sample is found with positive results contaminated with Coliform bacteria, namely 11 samples with a percentage of 78.6% and negative results of 3 samples with a percentage of 21.4%. Based on the results, it can be concluded that a positive result was found consisting of 1 sample of coliform fekal bacteria, in this case the bakteri Escherchia coli. bacteria, and 10 samples from the non-fecal coliform bacteria.

Antimicrobial activity of the crude extracts of Houttuynia cordata and Centella asiatica on some human gut microflora

C. asiatica and H. cordata plants are used as vegetable since time immemorable by the people of Assam. These are also consumed for their medicinal values. Plenty of information on the action of the plants extract on the pathogens are available, but could not retrieve any information on the non-pathogens of human guts. Therefore, an attempt has been made to find action of the extracts on few of the non-pathogens of human guts. The study shows that none of the plant extracts inhibit Bacillus coagulans, Bacillus mesentricus, and Lactobacillus sp, whereas a minimum inhibition were recorded in case of pathogens Escherchia coli, Staphylococcus aureus, Klebsiella pneumoniae and Shigella dysenteriae.