Punicalagin, a pomegranate polyphenol sensitizes the activity of antibiotics against three MDR pathogens of the Enterobacteriaceae.

BACKGROUND: Multidrug resistance (MDR) in the family Enterobacteriaceae is a perniciously increasing threat to global health security. The discovery of new antimicrobials having the reversing drug resistance potential may contribute to augment and revive the antibiotic arsenal in hand. This study aimed to explore the anti-Enterobacteriaceae capability of bioactive polyphenols from Punica granatum (P. granatum) and their co-action with antibiotics against clinical isolates of Enterobacteriaceae predominantly prevalent in South Asian countries. METHODS: The Kandhari P. granatum (Pakistani origin) extracts were tested for anti-Enterobacteriaceae activity by agar well diffusion assay against MDR Salmonella enterica serovar Typhi, serovar Typhimurium and Escherichia coli. Predominant compounds of active extract were determined by mass spectrometry and screened for bioactivity by agar well diffusion and minimum inhibitory concentration (MIC) assay. The active punicalagin was further evaluated at sub-inhibitory concentrations (SICs) for coactivity with nine conventional antimicrobials using a disc diffusion assay followed by time-kill experiments that proceeded with SICs of punicalagin and antimicrobials. RESULTS: Among all P. granatum crude extracts, pomegranate peel methanol extract showed the largest inhibition zones of 25, 22 and 19 mm, and the MICs as 3.9, 7.8 and 7.8 mg/mL for S. typhi, S. typhimurium and E. coli, respectively. Punicalagin and ellagic acid were determined as predominant compounds by mass spectrometry. In plate assay, punicalagin (10 mg/mL) was active with hazy inhibition zones of 17, 14, and 13 mm against S. typhi, S. typhimurium and E. coli, respectively. However, in broth dilution assay punicalagin showed no MIC up to 10 mg/mL. The SICs 30 µg, 100 µg, and 500 µg of punicalagin combined with antimicrobials i.e., aminoglycoside, ß-lactam, and fluoroquinolone act in synergy against MDR strains with % increase in inhibition zone values varying from 3.4 ± 2.7% to 73.8 ± 8.4%. In time-kill curves, a significant decrease in cell density was observed with the SICs of antimicrobials/punicalagin (0.03-60 µg/mL/30, 100, 500 µg/mL of punicalagin) combinations. CONCLUSIONS: The P. granatum peel methanol extract exhibited antimicrobial activity against MDR Enterobacteriaceae pathogens. Punicalagin, the bacteriostatic flavonoid act as a concentration-dependent sensitizing agent for antimicrobials against Enterobacteriaceae. Our findings for the therapeutic punicalagin-antimicrobial combination prompt further evaluation of punicalagin as a potent activator for drugs, which otherwise remain less or inactive against MDR strains.

Scorpion sting in Hafizabad, Pakistan: Facility based epidemiological study.

Scorpion envenomation is a widespread issue in tropical and subtropical countries. In the present study epidemiology of scorpion sting cases and potential treatment options in district Hafizabad, Pakistan are documented. Hafizabad city and the adjacent villages were selected for the data collection. Age, gender, time, site of scorpion sting on the body, scorpion sting symptoms, number of patients obtaining medical attention and the number of fatalities (if any) were noted. Data showed that scorpion envenomation often occurs in people between the ages of 16-47 years. There were more female victims (55%) and most of the scorpion sting incidences (55.49%) occurred between 07 PM and 02 AM Among body parts, both the hands (41.71%) and feet (48.57%) were more vulnerable to scorpion stings. In rural areas, the incidences of scorpion stings were higher (68.57%). The reported symptoms of scorpion stings include pain, hypertension, nausea and allergy. Out of all the victims, only 56.05% sought treatment from health care units as well as traditional medicine practitioners. However, others (43.94%) fully healed on their own without any treatment. People were using different types of plant-based materials and dead scorpions with mustard oil as a potent remedy against scorpion stings. Moreover, in the health care units there was no antivenom available and medical staff usually used common pain killers such as Xylocain and Lignocain for the treatment of scorpion sting.

Characterization and Evaluation of Silk Sericin-Based Hydrogel: A Promising Biomaterial for Efficient Healing of Acute Wounds.

The present study was aimed to prepare the potent silk sericin-based hydrogels in combination with plant extracts (curcumin and banana peel powder) and silver nanoparticles (AgNPs) to accelerate the acute wound healing process. Experimental excision wounds were created in mice by biopsy puncture, and the wound healing potential of silk sericin (2%)-based hydrogel and its combinations with curcumin (2%), banana peel powder (2%), and AgNPs (2%) was estimated by calculating the percent wound contraction, healing time, histology of skin tissues, and different biochemical tests. The results showed that the mice treated with sericin-based hydrogels showed significantly (P < 0.001) high percent wound contraction as compared to negative control, and wounds were healed in 11 days. The histological evaluation also showed that wounds covered with hydrogels were healed more than the uncovered wounds. Furthermore, the results of biochemical tests revealed that the treatment groups showed a significant (P < 0.001) decrease in the serum level of pro-inflammatory cytokines (IL-6). A significant (P < 0.001) increase in anti-inflammatory cytokines (IL-10) and anti-oxidant enzymes was observed in treatment groups. The highest wound healing potential was observed by sericin-based hydrogel containing banana peel powder, leaving behind the commercially available ointment polyfax (positive control). It can be concluded that the silk sericin-based hydrogels in combination with plant extract and AgNPs can be used as natural biomaterials in wound dressing for the rapid healing of acute wounds.

Diabetic wound healing potential of silk sericin protein based hydrogels enriched with plant extracts.

The complications associated with diabetic wounds make their healing process prolonged. Hydrogels could be ideal wound dressings therefore present research was conducted to prepare silk sericin (an adhesive protein polymer) based hydrogels in combination with plant extracts and to evaluate its effectiveness against wound healing process in mice with alloxan induced diabetes. Excision wounds were formed via a biopsy puncture (6 mm). Experimental hydrogels were prepared and applied topically on the diabetic wounds. All the hydrogel treatment groups showed significantly higher (P < 0.001) percent wound contraction from day 3 to day 11 as compared to the negative diabetic control group. The serum level of anti-inflammatory cytokine (Interleukin-10) and tissue inhibitor metalloproteinase (TIMP) was significantly higher (P < 0.001), while the level of pro-inflammatory cytokines (tumor necrosis factor-α, Interleukin-6) and matrix metalloproteinases (MMP-2, MMP-9) was significantly lower (P < 0.001) in hydrogels treatment groups as compared to diabetic control group. Although all the hydrogels showed effective results, however the best results were shown by 4 % sericin+4 % banyan+4 % onion based hydrogel. It can be concluded that Sericin based hydrogel enriched with banyan and onion extracts can be used as an effective remedy for the treatment of diabetic wounds due to their high healing and regenerative properties.

Exogeneous selenium enhances anthocyanin synthesis during grain development of colored-grain wheat.

Anthocyanins and selenium (Se) play critical roles in antioxidant, anticancer, antibacterial, and antiviral treatments. Previous studies indicate that colored-grain wheat accumulates more Se than regular wheat, and Se synergistically promotes anthocyanin synthesis. However, the mechanism through which Se regulates anthocyanin synthesis remains unclear. We studied anthocyanin accumulation during the grain-filling stage of colored-grain wheat development by employing transcriptomics and metabolomics. We show that Se biofortification increased the concentrations of Se, anthocyanin, chlorophyll a and b, and carotenoids in colored-grain wheat. Genes related to biosynthesis of anthocyanins, phenylpropanoids biosynthesis, and flavonoids biosynthesis were significantly upregulated after Se treatment, which led to the accumulation of anthocyanin metabolites in colored-grain wheat. Genetic alterations in the expression profiles of several genes and transcription factors were observed, which slowed down lignin and proanthocyanidin biosynthesis and accelerated anthocyanin synthesis. Our results deepen the understanding of anthocyanin metabolism in Se-treated colored-grain wheat, which will likely promote harvest of these varieties.

Efficacy and Safety of Prunus mume and Choline in Patients with Nonalcoholic Fatty Liver Disease.

Aim: The primary objectives of this study include evaluating changes in lipid profile and liver enzyme levels in nonalcoholic fatty liver disease (NAFLD) patients receiving Prunus mume and choline supplementation (Revolic). Materials and methods: Two-hundred patients were recruited from the hepatology outpatient department of a public hospital between January and June 2023. Patients who had confirmed diagnosis of NAFLD, proven with ultrasound (US) followed by biopsy or US alone with age >18 years were included in this study. The study variables were fasting blood sugar, cholesterol levels, low-density lipoprotein (LDL), triglyceride, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase levels (GGT). All investigations were conducted and compared between baseline, 12 and 24 weeks following treatment. Results: The mean age of all participants was 40.49 ± 10.59 years with 34 males and 166 females. The mean cholesterol levels were reduced to 179.86 ± 35.63 mg/dL from the mean baseline of 197.57 ± 42.52 mg/dL (p = 0.001). There was also a statistically significant difference found between baseline and posttreatment levels of LDL and triglyceride (p < 0.001). The ATL levels were also reduced from baseline 44.91 ± 32.40 U/L to 44.25 ± 30.66 and 41.06 ± 22.15 U/L between 12 and 24 weeks after treatment respectively. There was a statistically significant reduction in ATL, AST, and GGT levels from baseline with p-value < 0.001. Conclusion: The combination of P. mume and choline (Revolic) gives promising results with a significant reduction in lipid profile and liver enzymes. Clinical significance: The combination of P. mume and choline can be considered a reliable option for the management of NAFLD due to its efficacy and safety at 24 weeks after treatment as evident in the present study. How to cite this article: Butt N, Masood M, Ali A. Efficacy and Safety of Prunus mume and Choline in Patients with Nonalcoholic Fatty Liver Disease. Euroasian J Hepato-Gastroenterol 2023;13(2):124-127.

Iron oxide nanoparticles and selenium supplementation improve growth and photosynthesis by modulating antioxidant system and gene expression of chlorophyll synthase (CHLG) and protochlorophyllide oxidoreductase (POR) in arsenic-stressed Cucumis melo.

Current research reveals the positive role of iron oxide nanoparticles (IONPs) and selenium (Se) in extenuation of arsenic (As) induced toxicity in Cucumis melo. C. melo plants grown in As spiked soil (20 mg kg-1 As) showed reduced growth, chlorophyll (Chl) content, photosynthetic rate, stomatal conductivity and transpiration. On the other hand, the alone applications of IONPs or Se improved growth and physiochemical parameters of C. melo plants. Additionally, exogenous application IONPs and Se synergistically improved the activity of antioxidative enzymes and glyoxalase system in C. melo plants. In addition, the collective treatment of IONPs and Se reduced As uptake, enhanced rate of photosynthesis and increased gas exchange attributes of C. melo plants under As stress. Interactive effect of IONPs and Se regulated reduced glutathione (GSH), oxidized glutathione (GSSG) and ascorbate (AsA) content in C. melo plants exposed to As-contaminated Soil. IONPs and Se treatment also regulated expression of respiratory burst oxidase homologue D (RBOHD) gene, chlorophyll synthase (CHLG) and protochlorophyllide oxidoreductase (POR). Therefore, the combined treatment of IONPs and Se may enhance the growth of crop plants by alleviating As stress.

Detection of synergistic antimicrobial resistance mechanisms in clinical isolates of Pseudomonas aeruginosa from post-operative wound infections.

Infections caused by carbapenem-resistant Pseudomonas aeruginosa are life-threatening due to its synergistic resistance mechanisms resulting in the ineffectiveness of the used antimicrobials. This study aimed to characterize P. aeruginosa isolates for antimicrobial susceptibility, biofilm formation virulence genes, and molecular mechanisms responsible for resistance against various antimicrobials. Out of 700 samples, 91 isolates were confirmed as P. aeruginosa which were further classified into 19 non-multidrug-resistant (non-MDR), 7 multidrug-resistant (MDR), 19 extensively drug-resistant (XDR), and 8 pan drug-resistant (PDR) pulsotypes based on standard Kirby Bauer disc diffusion test and pulse field gel electrophoresis. In M9 minimal media, strong biofilms were formed by the XDR and PDR pulsotypes as compared to the non-MDR pulsotypes. The virulence genes, responsible for the worsening of wounds including LasB, plcH, toxA, and exoU, were detected among all MDR, XDR, and PDR pulsotypes. Carbapenemase activity was phenotypically detected in 45% pulsotypes and the responsible genes were found as blaGES (100%), blaVIM (58%), blaIMP (4%), and blaNDM (4%). Real-time polymerase chain reaction showed the concomitant use of multiple mechanisms such as oprD under-expression, enhanced efflux pump activity, and ampC overexpression in the resistant isolates. Polymyxin is found as the only class left with more than 80% susceptibility among the isolates which is an alarming situation suggesting appropriate measures to be taken including alternative therapies. KEY POINTS: • Multidrug-resistant P. aeruginosa isolates formed stronger biofilms in minimal media. • Only polymyxin antimicrobial was found effective against MDR P. aeruginosa isolates. • Under-expression of oprD and overexpression of ampC were found in resistant isolates.

ω-3 Ethyl ester results in better cognitive function at 12 and 30 months than control in cognitively healthy subjects with coronary artery disease: a secondary analysis of a randomized clinical trial.

BACKGROUND: Omega-3 (n-3) fatty acids have shown benefit in cognitively impaired subjects, but the effect on cognitively healthy older subjects is unclear. OBJECTIVES: Our aim was to determine if long-term, high-dose ω-3 ethyl esters, EPA (20:5n-3) and DHA (22:6n-3), prevent deterioration of cognitive function in cognitively healthy older adults. METHODS: A total of 285 subjects with stable coronary artery disease (CAD) on statin treatment were randomly assigned to 3.36 g EPA and DHA or none (control) for 30 mo. Cognitive function was assessed in all 285 subjects at baseline and in 268 and 250 subjects who returned at 12- and 30-mo follow-up, respectively, with neuropsychological testing as a prespecified secondary outcome. A completer's analysis, along with a sensitivity analysis carrying forward the last observation, was performed. RESULTS: Over the 30-mo period, subjects randomly assigned to EPA and DHA had significantly better scores than control for verbal fluency, language, and memory (mean: 1.08; 95% CI: 0.25, 1.91; P = 0.011) and 2 tests of visual-motor coordination (mean: -2.95; 95% CI: -5.33, -0.57; P = 0.015 and mean: -9.44; 95% CI: -18.60, -0.30; P = 0.043, respectively). The better scores for EPA and DHA were due to an improvement at 12 mo compared with baseline in verbal fluency, language, and memory (P = 0.047) and 2 tests of visual-motor coordination (P = 0.033 and P < 0.001, respectively), whereas control had no change. Post hoc analyses indicated no difference by age, sex, or diabetes status. CONCLUSIONS: Cognitively healthy older adults with stable CAD randomly assigned to high-dose EPA and DHA had improved cognitive function over a 30-mo period compared with control. These findings may be especially important for CAD patients because CAD is a risk factor for cognitive decline.This trial was registered at clinicaltrials.gov as NCT01624727.

Plant growth promoting rhizobacteria alleviates drought stress in potato in response to suppressive oxidative stress and antioxidant enzymes activities.

Maintenance of plant physiological functions under drought stress is normally considered a positive feature as it indicates sustained plant health and growth. This study was conducted to investigate whether plant growth-promoting rhizobacteria (PGPR) Bacillus subtilis HAS31 has potential to maintain potato growth and yield under drought stress. We analyzed trends of chlorophyll concentration, photosynthesis process, relative water content, osmolytes, antioxidants enzymes and oxidative stress, relative growth rate, tuber and aboveground biomass production in two potato varieties, Santae (drought-tolerant) and PRI-Red (drought-sensitive). Plants of both genotypes were treated with 100 g of HAS31 inoculant at 10 days after germination and exposed to different soil relative water contents (SRWC), including 80 ± 5% (well watered), 60 ± 5% (moderate stress) and 40 ± 5% SRWC (severe stress) for 7 days at tuber initiation stage (30 days after germination). The drought stress reduced plant relative growth rate, biomass production, leaf area, number of leaves and tubers, tuber weight, and final yield. The drought-stressed plants showed decline in chlorophyll contents, membrane stability, leaf relative water contents and photosynthetic rate. Under drought stress, enzymatic activity of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD), contents of total soluble sugars, soluble proteins and proline increased. The application of PGPR reduced the impact of drought and maintained higher growth and physio-chemical traits of the plants. The plants with PGPR application showed higher relative growth rate, dry matter production, leaf area, number of tubers, tuber weight and yield as compared to plants without PGPR. The PGPR-HAS31 treated plants maintained higher photosynthetic process, contents of chlorophyll, soluble proteins, total soluble sugars, and enzymatic activities of CAT, POD and SOD as compared to plants without PGPR. The results of the study suggest that plant growth regulators have ability to sustain growth and yield of potato under drought stress by maintaining physiological functions of the plants.

Ni tolerance and its distinguished amelioration by chelating agents is reflected in root radius of B. napus cultivars.

The negative effect of excess nickel (Ni) on plants is well investigated but there is only little information on its influence on root anatomy and a possible amelioration by chelating agents. In this study, we utilized light microscopy to observe anatomical changes in canola (Brassica napus) roots and investigated the element content by X-ray microanalysis. Ni-tolerant (Con-II) and Ni-sensitive cultivars (Oscar) were selected for this purpose. The plants were treated with 30 ppm NiSO4. Then, citric acid and ethylene-diamine-tetra-acetic acid (EDTA) (alone or in combination) were applied to observe the influence of chelating agents in metal stress amelioration. Ni treatment led to significant swelling of the roots in the Con-II variety as compared to the cultivar Oscar. Application of EDTA reduced the root radius of Con-II plants and this effect for Ni tolerance is discussed. According to X-ray microanalyses, Ni ions were more dispersed in the sensitive cultivar as indicated by metal adsorption to the cell wall. We investigate the hypothesis that an enhanced capacity of binding metals to the cell wall allows the plants to tolerate more heavy metals.

Halotolerant plant-growth promoting rhizobacteria modulate gene expression and osmolyte production to improve salinity tolerance and growth in Capsicum annum L.

Some rhizobacteria have demonstrated a noteworthy role in regulation of plant growth and biomass production under biotic and abiotic stresses. The present study was intended to explicate the ameliorative consequences of halotolerant plant growth-promoting rhizobacteria (HPGPR) on growth of capsicum plants subjected to salt stress. Salt stress was ascertained by supplementing 1 and 2 g NaCl kg-1 soil. The HPGPR positively invigorated growth attributes, chlorophyll, protein contents, and water use efficiency (WUE) of supplemented capsicum plants under salinity stress conditions. Bacillus fortis strain SSB21 caused highest significant increase in shoot length, root length, and fresh and dry biomass production of capsicum plants grown under saline conditions. This multi-trait bacterium also increased biosynthesis of proline and up-regulated the expression profiles of stress related genes including CAPIP2, CaKR1, CaOSM1, and CAChi2. On the other hand, B. fortis strain SSB21 inoculated plants exhibited reduced level of ethylene, lipid peroxidation, and reactive oxygen species (ROS). All these together contribute to activate physiological and biochemical processes involved in the mitigation of the salinity induced stress in capsicum plants.

The beneficial role of potassium in Cd-induced stress alleviation and growth improvement in Gladiolus grandiflora L.

Heavy metal contaminated agricultural soils are one of the most important constraints for successful cultivation of crops. The current research was conducted to evaluate the role of potassium (K) on plant growth and amelioration of cadmium (Cd) stress in Gladiolus grandiflora under greenhouse conditions. G. grandiflora corms were sown in media contaminated with 0 (C), 50 (Cd50) and 100 (Cd100) mg Cd kg-1 soil. The plants growing in Cd-contaminated media exhibited reduced gas exchange attributes, chlorophyll (Chl) contents, vegetative and reproductive growth as compared to control. The plants raised in Cd contaminated media showed reduced nutrition yet higher Cd contents. However, supplementation of 60 mg Kg-1 K in treated plants (C+K, Cd50+K and Cd100+K) improved quantity of total soluble protein and proline (Pro) along with activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) under Cd stress. Similarly, K supplementation reduced the level of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in treated plants. Potassium supplemented plants exhibited better vegetative and reproductive growth. The improved stress tolerance in K supplemented plants was attributed to the reduced quantity of MDA and H2O2, enhanced synthesis of protein, proline, phenols, flavonides and improved activity of antioxidant enzymes. The present research supports the application of K for alleviation of Cd stress in G. grandiflora.

Laparoscopic insertion of gastric electrodes for electrical stimulation.

Gastric electrical stimulation can provide symptomatic relief for patients with refractory gastroparesis. Traditionally, these wires are placed through a midline laparotomy. This paper describes and illustrates, in detail, the laparoscopic technique for successful implantation. Thirty-one consecutive patients from October 2003 to March 2005 underwent laparoscopic insertion of gastric stimulating wires for gastroparesis. Twenty-six patients were female. Four laparoscopic ports were used to insert a pair of electrodes. Anterior, cephalad retraction of the gastric wall is critical for accurate seromuscular placement of gastric leads. Intraoperative endoscopy was used to verify the seromuscular placement of the leads. Both leads were secured to a subcutaneous generator and electrical parameters were immediately established in the operating room. Patient demographics, operative details, and postoperative morbidities were recorded. All procedures were completed laparoscopically. The mean operative time was 114.4 +/- 20.9 minutes (range, 95-140). No perioperative mortality occurred. Two patients developed cellulitis at the generator site postoperatively and oral antibiotics were prescribed for one week postoperatively. No hardware was removed. Two patients had their generators repositioned due to pain at the pocket site. Gastric electrical stimulation is a novel treatment modality for patients with refractory gastroparesis and can be accomplished safely via laparoscopy. Laparoscopic insertion is successful even in patients with prior surgery and intact gastrointestinal tubes. Long-term follow-up and the current prospective multicenter trial continue to assess the efficacy of this treatment modality.