Factor affecting Agrobacterium-mediated transformation of rice chitinase gene in Solanum tuberosum L.

Potato is a very important food crop and is adversely affected by fungus. Agrobacterium-mediated transformation can play an important role in the improvement of potato. The present study was conducted to optimize the different factors affecting Agrobacterium-mediated transformation of chitinase gene. Nodes were used as explant of potato cultivars Desiree and Sh-5. Agrobacterium tumefacien strain LBA4404 harboring a pB1333-EN4-RCG3 plasmid having chitinase gene and selectable marker hygromycin gene under the control of the CaMV 35S promoter was used. The parameters optimized for the potato transformation includes co-cultivation time, cefotaxime concentration, and days to pre-selection. The results show that the infection time (2 min) gave best mean value of transformation efficiency, that is, 2.9 and 2.1 in Desiree and Sh-5, respectively. The over growth of Agrobacterium were controlled with a concentration of 200 mg/l of cefotaxime and obtained maximum mean value of transformation efficiency of 3.38 and 3.10 in Desiree and Sh-5, respectively. The pre-selection period seven days prior to selection were considered effective for regeneration of explants and high transformation efficiency. The high mean value of regeneration (3.08 and 2.82) and transformation efficiency (3.00 and 2.60) was observed in Desiree and Sh-5, respectively. The putative transgenic plants were analyzed through PCR by using RCG-3 specific primers.Key words: Agrobacterium, potato, transformation, rice chitinase gene, factors

Comparative analysis of hyoscine in wild-type and in vitro grown Datura innoxia by high performance liquid chromatography

Purpose: To compare the hyoscine contents of Datura innoxia plant grown in the wild and that grown in vitro.Methods: For callus induction from leaf explants, 15 combinations of 2,4-dichlorophenoxy acetic acid, gibberellic acid, benzyl amino purine and kinetin were used. For regeneration of shoots, 8 combinations of BAP, kinetin and indole butyric acid (IBA) were tested. Different concentrations of IBA were used for rooting. The hyoscine contents of the wild and in vitro samples was initially identified by TLC and subsequently quantified by high performance liquid chromatography using hyoscine N-butyl bromide as standard.Results: For callus induction, 2,4-D at 2 mg/L produced maximum calli (96 %). Maximum shoots regeneration was obtained with 2 mg/L BAP and 1 mg/L kinetin. IBA at 0.5 mg/L induced good rooting. Substantial amount of hyoscine was detected in the extracts of in vitro grown plants through TLC and HPLC. Wild root, stem and leaves exhibited higher amounts (approx. 2 μg/mL) of hyoscine than the parts of in vitro grown plants. Green callus grown in vitro contained a maximum concentration of hyoscine (1.01 μg/mL) followed by leaf (0.82 μg/mL) and brown callus (0.432 μg/mL).Conclusion: The callus cultures of D. innoxia are capable of yielding good amounts of hyoscine, and therefore can be exploited to increase hyoscine production using chemical/hormone treatments on a large scale. Thus, this study provides a sustainable and efficient way for the sustainable production of a natural hyoscine product via in vitro-grown D. innoxia cultures.Keywords: Datura innoxia, Hyoscine, Callus, Hyoscine, Explant, Gibberellic acid, Benzyl amino purine, Kineti

Proprotein convertase subtilisin/Kexin type-9 (PCSK-9) inhibitors induced liver injury - a retrospective analysis.

Background: Proprotein convertase subtilisin/Kexin type 9 (PCSK-9) inhibitors induced liver dysfunction in patients with or without previous liver injury, and this is not well discussed in the previous literature. Methods: A total sample of 202 patients were retrospectively reviewed at the University of Missouri, Kansas City, from the year 2015 to 2018 based on predefined selection criteria. Inclusion criteria involved patients with dyslipidemia, with or without PCSK-9 inhibitors, liver function tests and lipid profile at baseline and at a mean of 6-month follow-up. The variables, including age, gender, and confounding factors like other medications (statin, oral antidiabetic, and antihypertensive) induced, or chronic secondary liver diseases causing liver injury were taken into consideration. Exclusion criteria included patients without dyslipidemia. Results: The mean age of the study population was 64 ± 11 years (63% males and 37% females). The lipid profile including triglyceride and cholesterol levels during 6-month followup visit showed a mean of 184 ± 260 and 163 ± 50 mg/dL as compared to that at baseline of 227 ± 603 and 181 ± 70 mg/dL, respectively. In terms of clinical efficacy, a 6-month follows-up showed a drop in triglyceride and cholesterol levels by 38 and 15 mg/dL, respectively. A liver function test at 6 months in patients taking PCSK-9 inhibitors showed an increase in alanine transaminase (ALT) and aspartate transaminase (AST) by 5.8 mg/dL (p = 0.037) and 6.2 mg/dL (p = 0.008), respectively, from baseline values. Conclusion: PCSK-9 inhibitors should be used cautiously with a follow-up liver function test

Prevalence of work related musculoskeletal disorders among physiotherapists of Multan

Background: Work related musculoskeletal disorders are increasing day by day in hospitals and clinics due to high demand activities. These disorders decline the efficiency of therapists and result in social as well as economic losses. These injuries mostly occur due to abnormal postures and poor ergonomics. Aim and objectives: The purpose of this study was to rule out pain which disturbs the daily living of physiotherapists due to work related musculoskeletal disorder. This study also helps physiotherapists to improve their skills, technique, posture, ergonomics and all the other factors that are related to these disorders. Method: Survey was performed among physiotherapists that are performing duties in government hospitals, private clinics and all the other settings in Multan. This study included 100 physiotherapists of Multan. Self-design questionnaire as well as a Nordic questionnaire was used to collect data about WRMSK disorders. Data was gathered and was observed statistically. Results: 89% of physiotherapist were having had work related musculoskeletal disorder & 11% had no complaint, 75% female and 25% male & 23 to 50 years age population data was taken & the study observed that novice practitioner are at the increased risk of developing work-related musculoskeletal disorder & 45% were non specialized and 55% were specialized. The most commonly affected regions were upper back, low back and hip joint respectively. Conclusion: Upper back pain, lower back pain and hip pain were the most common complications among physiotherapists. Manual therapy shows association with the prevalence of MSK disorder

Protocol optimization for deoxyribonucleic acid (DNA) extraction from dried, fresh leaves, and seeds of groundnut (Arachis hypogaea L.)

Consistent isolation of best quality deoxyribonucleic acid (DNA) from peanut (Arachis hypogaea L.) is particularly problematic due to the presence of phenolic compounds and polysaccharides. Inconsistencies in extraction results can be attributed to the age and growth stages of the plant material analyzed. Mature leaves have higher quantities of polyphenols, tannins and polysaccharides that can contaminate DNA during isolation. In this study, we used fresh and dried leaves as well as seeds for optimization of high quality DNA isolation protocols from A. hypogaea. The DNA extracted with three different methods cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), and cesium chloride (CsCl) density gradient) were comparatively studied by polymerase chain reaction (PCR) analysis in terms of quantity and quality. High quality genomic DNA was obtained from fresh leaves by modified CTAB methods. The DNA obtained ranged from 1 to 2.5 ng/μl. DNA obtained by this method was strong and reliable showing its compatibility for simple sequence repeat (SSR) analyses. The SDS based methodology give large quantities of DNA contaminated with polysaccharides. Fresh leaves also gave best result in SDS method. The quantity and quality of DNA obtained was very poor in all the tested methods in case of dried leaf tissues. The current protocol will probably be useful for the extraction of high-molecular weight DNA from other plant materials containing large amounts of secondary metabolites and essential oils.Key words: Polysaccharides, polyphenols, tannins, cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), cesium chloride (CsCl), secondary metabolites, SSR

Identifying Key Genes of Liver Cancer by Using Random Forest Classification

Liver cancer is considered as one of the most deadly cancer. To devise a treatment which is helpful to eradicate, it is inevitable to identify potential biomarkers which are very important in the development of liver cancer. To identify the pathways and key genes we use different enrichment analysis techniques such as pathway analysis and functional analysis. To identify biomarkers we constructed a network which is named as protein protein interaction network to analyse by selecting different network nodes. Our results show that we identified those biomarkers like ESR1 and TOP2 successfully which are potential biomarkers for liver cancer. In addition to that our method can be applied to other different datasets which are for different diseases to choose key genes

Promising Compounds for Treatment of Covid-19

The study spans over-identification of phytochemicals suited for the treatment of COVID-19. The study focuses on the chemical that has a tendency to bind with Human ACE2 protein and two of the main Sars-Cov-2 proteins which are the Spike protein and the RNA-directed RNA polymerase (RdRp) protein. After screening a large list of phytochemicals two of the compound i.e. Kansuinine B and Digitoxin were found to have promising traits for the treatment of COVID-19. Both the compounds have been in use for centuries. Digitoxin was extracted from Foxglove seeds in the 18th century for heart-related illnesses. Kansuinine B originates from a Chinese herb Euphorbia Kansui (E. Kansui) E. Kansui has been widely used in herbal medicine for a multitude of illnesses including lungs related diseases. Studies also show that it has the ability to suppress cytokine response through the expression of the SOCS3 gene. In-silico simulations show that both these compounds have a better affinity and binding properties with these three proteins as compared to many other drugs under trial for COVID-19 like Remdesivir, Ritonavir, Famotidine, Camostat Mesylate, and Hesperidin. A treatment based on the combination of both compounds can be very effective. Any self-medication of both the compounds is highly discouraged as misuse of both the compounds can be very harmful

A Novel De-Ghosting Image Fusion Technique for Multi-Exposure, Multi-Focus Images Using Guided Image Filtering

In this paper, a novel de-ghosting image fusion technique is presented, which enhances the quality of low dynamic range images using multi-level exposures taken from the ordinary camera and also removes the ghosting artifact. In the proposed algorithm, first, the source images, taken under different exposure settings, are decomposed into base and detail layers using two-scale decomposition. The base and detail layers contain small and large-scale variation details of the source images, respectively. The Laplacian-of-Gaussian filter is applied to the source images to get the edge information. Afterward, the saliency map of the edges is computed. To remove the ghosting artifacts, a weight matrix is calculated by applying the median filter on the histogram equalized source images. The weight matrix is combined with the saliency map to generate more accurate weights. The separate weights for the base and detail layers are calculated using guided image filters. Finally, the base and detail layers' weights are fused with the source images to generate a vivid and enhanced image without any artifacts. The proposed technique is evaluated both qualitatively and quantitatively. The comparison of our technique in terms of Yang's Metric (QY), Quality Mutual Information (QMI), Gradient-based Fusion Metric (QG) and Chen Blum's Metric (QCB) with other state-of-the-art techniques proves that the proposed technique outperforms existing techniques