Protein Profiling of Arabidopsis Roots Treated With Humic Substances: Insights Into the Metabolic and Interactome Networks

Background and Aim: Humic substances (HSs) influence the chemical and physical properties of the soil, and are also known to affect plant physiology and nutrient uptake. This study aimed to elucidate plant metabolic pathways and physiological processes influenced by HS activity. Methods: Arabidopsis roots were treated with HS for 8 h. Quantitative mass spectrometry-based proteomics analysis of root proteins was performed using the iTRAQ (Isobaric Tag for Relative and Absolute Quantification) technique. Out of 902 protein families identified and quantified for HS treated vs. untreated roots, 92 proteins had different relative content. Bioinformatic tools such as STRING, KEGG, IIS and Cytoscape were used to interpret the biological function, pathway analysis and visualization of network amongst the identified proteins. Results: From this analysis it was possible to evaluate that all of the identified proteins were functionally classified into several categories, mainly redox homeostasis, response to inorganic substances, energy metabolism, protein synthesis, cell trafficking, and division. Conclusion: In the present study an overview of the metabolic pathways most modified by HS biological activity is provided. Activation of enzymes of the glycolytic pathway and up regulation of ribosomal protein indicated a stimulation in energy metabolism and protein synthesis. Regulation of the enzymes involved in redox homeostasis suggest a pivotal role of reactive oxygen species in the signaling and modulation of HS-induced responses

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

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

Brugada syndrome unmasked by fever: a comprehensive review of literature

Background: The Brugada pattern is identified on the EKG by a coved ST-segment elevation accompanied by a negative T wave in the early precordial leads in the absence of a cardiac structural abnormality. Brugada pattern and Brugada syndrome should be differentiated, as the latter is associated with an increased risk of sudden cardiac death. Methods: The literature was searched using multiple databases to identify all the articles on Brugada pattern. Data were screened and analyzed by independent authors. Results: Sixty articles, comprising 71 patients, were included in the study. The mean age of patients was 42.6 years, with a higher prevalence of Brugada pattern in men (83%) than women (17%). The most frequent findings associated with Brugada pattern was fever (83%). Other less common presentations included cough (21%), sore throat (10%), syncope (18%), abdominal pain (8%), and chest pain (7%). Comorbidities included pneumonia (30%), upper respiratory tract infections (14%) and smoking (14%). Among treatment modalities, 39% of patients had ICD placement performed, 44% received antibiotics, while 14% had supportive care. Adenosine was given to 3% of patients, while other antiarrhythmics like milrinone, amiodarone, sotalol, procainamide, flecainide, and nitroglycerin were given to 1% of patients. Most patients with Brugada syndrome had a satisfactory outcome, with only 4% mortality rate(WHAT ABOUT THE OTHER 11%?). Out of the 71 patients, 3% had persistent Brugada patterns, while 86% of patients recovered completely. There was no significant effect of ICD on mortality or Brugada pattern resolution (p 0.37). Conclusion: Our study shows that fever is the main reason for unmasking the Brugada pattern in patients with this channelopath

Lymphocyte-to-C-Reactive Protein Ratio: A Novel Predictor of Adverse Outcomes in COVID-19.

Background: Systemic inflammation elicited by a cytokine storm is considered a hallmark of coronavirus disease 2019 (COVID-19). This study aims to assess the validity and clinical utility of the lymphocyte-to-C-reactive protein (CRP) ratio (LCR), typically used for gastric carcinoma prognostication, versus the neutrophil-to-lymphocyte ratio (NLR) for predicting in-hospital outcomes in COVID-19. Methods: A retrospective cohort study was performed to determine the association of LCR and NLR with the need for invasive mechanical ventilation (IMV), dialysis, upgrade to an intensive care unit (ICU) and mortality. Independent Results: The mean age for NLR patients was 63.6 versus 61.6, and for LCR groups, it was 62.6 versus 63.7 years, respectively. The baseline comorbidities across all groups were comparable except that the higher LCR group had female predominance. The mean NLR was significantly higher for patients who died during hospitalization (19 vs. 7, P ≤ 0.001) and those requiring IMV (12 vs. 7, P = 0.01). Compared to alive patients, a significantly lower mean LCR was observed in patients who did not survive hospitalization (1,011 vs. 632, P = 0.04). For patients with a higher NLR (\u3e 10), the unadjusted odds of mortality (odds ratios (ORs) 11.0, 3.6 - 33.0, P \u3c 0.0001) and need for IMV (OR 3.3, 95% CI 1.4 - 7.7, P = 0.008) were significantly higher compared to patients with lower NLR. By contrast, for patients with lower LCR (\u3c 100), the odds of in-hospital all-cause mortality were significantly higher compared to patients with a higher LCR (OR 0.2, 0.06 - 0.47, P = 0.001). The aORs controlled for baseline comorbidities and medications mirrored the overall results, indicating a genuinely significant correlation between these biomarkers and outcomes. Conclusions: A high NLR and decreased LCR value predict higher odds of in-hospital mortality. A high LCR at presentation might indicate impending clinical deterioration and the need for IMV

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

Diagnostic Accuracy of Cardiac Magnetic Resonance Versus Fractional Flow Reserve: A Systematic Review and Meta-Analysis.

Background: Fractional flow reserve (FFR) is considered the gold standard for diagnosis of coronary artery disease (CAD). Stress Cardiac magnetic resonance (SCMR) has been recently gaining traction as a non-invasive alternative to FFR. Methods: Studies comparing the diagnostic accuracy of SCMR versus FFR were identified and analyzed using Review Manager (RevMan) 5.3 and Stata software. Results: A total of 28 studies, comprising 2,387 patients, were included. The pooled sensitivity and specificity for SCMR were 86% and 86% at the patient level, and 82% and 88% at the vessel level, respectively. When the patient-level data were stratified based on the FFR thresholds, higher sensitivity and specificity (both 90%) were noted with the higher cutoff (0.75) and lower cutoff (0.8), respectively. At the vessel level, sensitivity and specificity at the lower FFR threshold were significantly higher at 88% and 89%, compared to the corresponding values for higher cutoff at 0.75. Similarly, meta-regression analysis of SCMR at higher (3T) resolution showed a higher sensitivity of 87% at the patient level and higher specificity of 90% at the vessel level. The highest sensitivity and specificity of SCMR (92% and 94%, respectively) were noted in studies with CAD prevalence greater than 60%. Conclusions: SCMR has high diagnostic accuracy for CAD comparable to FFR at a spatial resolution of 3T and an FFR cut-off of 0.80. An increase in CAD prevalence further improved the specificity of SCMR