Therapeutic Values of Earthworm Species Extract from Azad Kashmir as Anticoagulant, Antibacterial, and Antioxidant Agents.

AIMS: Current research aimed to explore the therapeutic values of different earthworms as antibacterial, anticoagulant, and antioxidant agents. METHODS: Ten different earthworms, i.e., Amynthas corticis, Amynthas gracilis, Pheretima posthuma, Eisenia fetida, Aporrectodea rosea, Allolobophora chlorotica, Aporrectodea trapezoides, Polypheretima elongata, Aporrectodea caliginosa, and Pheretima hawayana, were collected and screened for biological activities. Antibacterial effect analysis of earthworm species was done against fourteen bacterial pathogens, i.e., Escherichia coli, Serratia marcescens, Streptococcus pyogenes, Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa (1), Salmonella typhimurium, Shigella flexneri, Enterobacter amnigenus, Serratia odorifera, Pseudomonas aeruginosa (2), Staphylococcus warneri, and Lactobacillus curvatus, via agar well diffusion, crystal violet, MTT, agar disc diffusion, and direct bioautography assays. Antioxidant potential was evaluated through ABTS and DPPH assays. Lipolytic, proteolytic, and amylolytic assays were done for lipase, protease, and amylase enzymes confirmation. In vitro anticoagulant effects were examined in the blood samples by measuring prothrombin time. RESULTS: Results revealed that all earthworm extracts showed the inhibition of all tested bacterial pathogens except P. aeruginosa (1), P. aeruginosa (2), S. warneri, and L. curvatus. The maximum zone of inhibition of E. coli was recorded as 14.66 ± 0.57 mm by A. corticis, 25.0 ± 0.0 mm by P. posthuma, 20.0 ± 0.0 mm by E. fetida, and 20.0 ± 0.0 mm by A. trapezoid. Cell proliferation, biofilm inhibition, the synergistic effect of extracts along with antibiotics, and direct bioautography supported the results of agar well diffusion assay. Similarly, P. hawayana, A. corticis, A. caliginosa, and A. trapezoids increase the prothrombin time more efficiently compared to other earthworms. A. corticis, A. gracilis, A. rosea, A. chlorotica, P. elongata, and A. trapezoides showed maximum DPPH scavenging potential effect. CONCLUSIONS: The coelomic fluid of earthworms possessed several bioactive compounds/enzymes/antioxidants that play an important role in the bacterial inhibition and act as anticoagulant agents. Therefore, the development of new therapeutic drugs from invertebrates could be effective and potential for the prevention of the emergence of multidrug-resistant bacteria.

Intra-Articular Piroxicam And Triamcinolone In A Ratmodel Of Osteoarthritis; Elevation And Comparison Of Chondroprotective Efficacy.

BACKGROUND: Osteoarthritis is the most common chronic degenerative joint disease. Definite treatment of osteoarthritis is still undiscovered. This study was designed to evaluate and compare the chondroprotective efficacy of piroxicam and triamcinolone in rat model of osteoarthritis. Methods: This laboratory based experimental study was conducted in Pharmacology Department, Army Medical College, Rawalpindi, from April-June 2019. Osteoarthritis was induced by medial meniscectomy and anterior cruciate ligament resection in knee joints of twenty-four rats. They were divided in three groups with eight rats in each. Group I, II and III were control, piroxicam and triamcinolone groups that were treated by intra articular saline, piroxicam and triamcinolone once weekly for four weeks respectively and then gait pattern was scored. Animals were euthanized thereafter and samples were taken for histopathological analysis. RESULTS: Comparison of gait score of control, piroxicam and triamcinolone groups exhibited a p-value of <0.01. Intergroup comparison of gait of group I and II, group I and III and group II and IV depicted pvalue of <0.001,0.013 and 0.013 respectively. Likewise histopathological comparison of control, piroxicam and triamcinolone groups showed p-value of <0.01. While Intergroup histopathological comparison of group I and II, group I and III and group II and IV showed p-value of <0.001, <0.001 and 0.008 respectively. Conclusion: Comparison of control group with treatment group proved chondroprotective efficacy of piroxicam and triamcinolone. On comparison of treatment groups, it was concluded that piroxicam has better chondroprotective efficacy as compared to triamcinolone.

ESIDE: A computationally intelligent method to identify earthworm species (E. fetida) from digital images: Application in taxonomy.

Earthworms (Crassiclitellata) being ecosystem engineers significantly affect the physical, chemical, and biological properties of the soil by recycling organic material, increasing nutrient availability, and improving soil structure. The efficiency of earthworms in ecology varies along with species. Therefore, the role of taxonomy in earthworm study is significant. The taxonomy of earthworms cannot reliably be established through morphological characteristics because the small and simple body plan of the earthworm does not have anatomical complex and highly specialized structures. Recently, molecular techniques have been adopted to accurately classify the earthworm species but these techniques are time-consuming and costly. To combat this issue, in this study, we propose a machine learning-based earthworm species identification model that uses digital images of earthworms. We performed a stringent performance evaluation not only through 10-fold cross-validation and on an external validation dataset but also in real settings by involving an experienced taxonomist. In all the evaluation settings, our proposed model has given state-of-the-art performance and justified its use to aid earthworm taxonomy studies. We made this model openly accessible through a cloud-based webserver and python code available at https://sites.google.com/view/wajidarshad/software and https://github.com/wajidarshad/ESIDE.

Postpartum haemorrhage: a preventable cause of maternal mortality.

OBJECTIVE: To assess the preventable predictors of severe postpartum haemorrhage and the adverse outcome associated with it. DESIGN: Observational study. PLACE AND DURATION OF STUDY: Department of Obstetrics and Gynaecology, Hayatabad Medical Complex, PGMI, Peshawar, form January to December 2001. PATIENTS AND METHODS: All the admitted patients who developed severe postpartum haemorrhage (>1500 ml) were included in the study. Clinical and sociodemographic data was obtained along with results of investigations to categorize the complications encountered. Odds ratio (OR) and 95% confidence intervals were determined. RESULTS: During the study period, 75 out of 4683 obstetrical admissions, developed severe postpartum haemorrhage (1.6 %). About 65% of the patients were admitted with some other complications including obstructed labour, antepartum haemorrhage and eclampsia. The risk factors were grand multiparity (OR=3.4), pre-eclampsia (OR=2.75), antepartum haemorrhage (OR=13.35), active labour of more than 10 hours (OR=46.92), twin delivery (OR=3.25), instrumental delivery (OR=8.62) and caesarean section (OR=9.74). Maternal mortality in these cases was 2.66% and residual morbidity being 40%. Birth attendant other than doctor and delivery outside the study unit were significantly associated with the adverse outcome in these patients. CONCLUSION: Maternal outcome associated with postpartum haemorrhage is a function of care given during labour and postnatal period with early diagnosis and management of the complication and its risk factors, being the key of good maternal outcome.

Phylogenetic and Comparative Sequence Analysis of Thermostable Alpha Amylases of kingdom Archea, Prokaryotes and Eukaryotes.

Alpha amylase family is generally defined as a group of enzymes that can hydrolyse and transglycosylase α-(1, 4) or α-(1, 6) glycosidic bonds along with the preservation of anomeric configuration. For the comparative analysis of alpha amylase family, nucleotide sequences of seven thermo stable organisms of Kingdom Archea i.e. Pyrococcus furiosus (100-105°C), Kingdom Prokaryotes i.e. Bacillus licheniformis (90-95°C), Geobacillus stearothermophilus (75°C), Bacillus amyloliquefaciens (72°C), Bacillus subtilis (70°C) and Bacillus KSM K38 (55°C) and Eukaryotes i.e. Aspergillus oryzae (60°C) were selected from NCBI. Primary structure composition analysis and Conserved sequence analysis were conducted through Bio Edit tools. Results from BioEdit shown only three conserved regions of base pairs and least similarity in MSA of the above mentioned alpha amylases. In Mega 5.1 Phylogeny of thermo stable alpha amylases of Kingdom Archea, Prokaryotes and Eukaryote was handled by Neighbor-Joining (NJ) algorithm. Mega 5.1 phylogenetic results suggested that alpha amylases of thermo stable organisms i.e. Pyrococcus furiosus (100-105°C), Bacillus licheniformis (90-95°C), Geobacillus stearothermophilus (75°C) and Bacillus amyloliquefaciens (72°C) are more distantly related as compared to less thermo stable organisms. By keeping in mind the characteristics of most thermo stable alpha amylases novel and improved features can be introduced in less thermo stable alpha amylases so that they become more thermo tolerant and productive for industry.