[Rapid determination of nine preservatives in tobacco flavor by three phase-hollow fiber-liquid phase microextraction-high performance liquid chromatography].

Tobacco flavors are extensively utilized in traditional tobacco products, electronic nicotine, heated tobacco products, and snuff. To inhibit fungal growth arising from high moisture content, preservatives such as benzoic acid (BA), sorbic acid (SA), and parabens are often incorporated into tobacco flavors. Nonetheless, consuming preservatives beyond safety thresholds may pose health risks. Therefore, analytical determination of these preservatives is crucial for both quality assurance and consumer protection. For example, BA and SA can induce adverse reactions in susceptible individuals, including asthma, urticaria, metabolic acidosis, and convulsions. Parabens, because of their endocrine activity, are classified as endocrine-disrupting chemicals. Despite extensive research, the concurrent quantification of trace-level hydrophilic (BA and SA) and hydrophobic (methylparaben, ethylparaben, isopropylparaben, propylparaben, butylparaben, isobutylparaben, and benzylparaben) preservatives in tobacco flavors remains challenging. Traditional liquid phase extraction coupled with high performance liquid chromatography (HPLC) often results in high false positive rates and inadequate sensitivity. In contrast, tandem mass spectrometry offers high sensitivity and specificity; however, its widespread application is limited by laborious sample preparation and significant operational costs. Therefore, it is crucial to establish a fast and sensitive sample pretreatment and analysis method for the nine preservatives in tobacco flavors. In this study, a method for the simultaneous determination of the nine preservatives (SA, BA and seven parabens) in tobacco flavor was established based on three phase-hollow fiber-liquid phase microextraction (3P-HF-LPME) technology combined with HPLC. To obtain the optimal pretreatment conditions, extraction solvent type, sample phase pH, acceptor phase pH, sample phase volume, extraction time, and mass fraction of sodium chloride, were examined. Additionally, the HPLC parameters, including UV detection wavelength and mobile phase composition, were refined. The optimal extraction conditions were as follows: dihexyl ether was used as extraction solvent, 15 mL sample solution (pH 4) was used as sample phase, sodium hydroxide aqueous solution (pH 12) was used as acceptor phase, and the extraction was carried out at 800 r/min for 30 min. Chromatographic separation was accomplished using an Agilent Poroshell 120 EC-C18 column (100 mm×3 mm, 2.7 µm) and a mobile phase comprising methanol, 0.02 mol/L ammonium acetate aqueous solution (containing 0.5% acetic acid), and acetonitrile for gradient elution. Under the optimized conditions, the nine target analytes showed good linear relationships in their respective linear ranges, the correlation coefficients (r) were ≥0.9967, limits of detection (LODs) and quantification (LOQs) were 0.02-0.07 mg/kg and 0.08-0.24 mg/kg, respectively. Under two spiked levels, the enrichment factors (EFs) and extraction recoveries (ERs) of the nine target analytes were 30.6-91.1 and 6.1%-18.2%, respectively. The recoveries of the nine target analytes ranged from 82.2% to 115.7% and the relative standard deviations (RSDs) (n=5) were less than 14.5% at low, medium and high levels. The developed method is straightforward, precise, sensitive, and well-suited for the rapid screening of preservatives in tobacco flavor samples.

A review of the key ingredients in industrial formulations of baby wet wipes.

The skin of newborns is classified as sensitive, with a higher risk of skin barrier disruption and irritation of a diapered area. Despite dermatologist recommendations to use only water and a cloth for cleaning, most of the population still relies on the comforts of modern parenting, which includes intensive daily usage of baby wet wipes. Novel baby formulations are designed following the concept of infant skin health, containing a gentle cleanser, suitable emollient, and buffer system enabling a slightly acidic pH value and they are free of ethyl alcohol. Thus, it is important to understand the chemical background of such a complex liquid formulation, with emphasis on its safety. In line with this, the present paper discusses the scientific background of various chemical compounds found in baby wipe formulations to improve the understanding of wet wipe designs and direct them toward more skin-friendly solutions.

Antimicrobial activities of natural flavonoids against foodborne pathogens and their application in food industry.

The application of natural alternatives as food preservatives has gained much attention due to the escalating negative perception of synthetic preservatives among consumers and the spread of drug-resistance foodborne pathogens. Natural flavonoids have the potential to be employed for food safety due to their antimicrobial properties against a wide range of foodborne pathogenic microorganisms. In this perspective, we reviewed the antimicrobial activities of natural flavonoids, the mechanism of action, as well as their application for food safety and quality. Various strategies for the incorporation of flavonoids into food products were highlighted, including direct addition to food formulations, encapsulation as micro or nanocarriers, and incorporation into edible or active films and coatings. Furthermore, we discussed the current challenges of industrial application of flavonoids, and proposed future trends to enhance their potential as natural preservatives. This review provides a theoretical foundation for the further development and application of flavonoids for food safety.

The chemical composition and the preservative, antimicrobial, and antioxidant effects of Thymus broussonetii Boiss. essential oil: an in vitro and in silico approach.

Introduction: The aim of this study was to evaluate the antioxidant, antimicrobial, and preservative efficacy of Thymus broussonetii Boiss. essential oil (EO) in a topically applied formulation using a challenge test. Methods: The essential oil was extracted from the aerial part of T. broussonetii using hydrodistillation, and the obtained EO was further analyzed by gas chromatography/mass spectrometry (GC/MS). The antioxidant effect of the EO was evaluated using three methods: the inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), ß-carotene-linoleic acid, and the ferric reducing antioxidant power (FRAP) methods. The antimicrobial activity and the minimum inhibitory concentration (MIC) of this EO were assayed by the disk-diffusion method and the broth microdilution method, respectively. The preservative efficacy of T. broussonetii EO was assayed at 1% and 2% (v/w) in a topical cream formulation using a challenge test against standard-specific microorganisms recommended by the European Pharmacopoeia. Furthermore, the identified phytochemical compounds were docked for their effect on nicotinamide adenine dinucleotide phosphate oxidase, human casein kinase 1 alpha 1 (CSNK1A1), glycogen synthase kinase 3, Staphylococcus aureus nucleoside diphosphate kinase, Escherichia coli beta-ketoacyl-[acyl-carrier protein] synthase, Pseudomonas aeruginosa LasR ligand-binding domain, and sterol 14-alpha demethylase (CYP51) from Candida albicans. The ADME/toxicity was predicted by analyzing the absorption, distribution, metabolism, and excretion parameters. Results and discussion: chemical composition of the EO revealed the presence of thymol (63.09%), p-cymene (11%), and γ-terpinene (8.99%) as the major components. The antioxidant assays revealed that the essential oil exhibited strong antioxidant activity, as indicated by the minimum inhibitory concentration IC50 (IC50 = 210 ± 0.3 µg/mL for the DPPH assay, IC50 = 145 ± 0.1 µg/mL for the ß-carotene assay, and IC50 = 84 ± 0.21 µg/mL for the FRAP assay) when compared to quercetin and butylated hydroxytoluene (BHT) as controls. The investigated essential oil exhibited important antimicrobial activity against all the tested microorganisms, and the MICs of the EO against bacteria and fungi were 0.02%-1%. Moreover, the EO of T. broussonetii evaluated at 2% (v/w) in a cream formulation succeeded in satisfying the A criteria for preservation efficacy against S. aureus, E. coli, and Aspergillus brasiliensis but exhibited less efficacy against P. aeruginosa (1.78 log reduction in the number of CFU/g after 7 days of evaluation) and C. albicans (1.09 log reduction in the number of CFU/g after 14 days of evaluation) when compared to the synthetic preservative phenoxyethanol 1% (v/w). In silico results showed that the antimicrobial activity of T. broussonetii EO is mostly attributed to thymol, terpinen-4-ol, and aromadendrene, while the antioxidant activity is attributed to thymol. These results indicate that the EO of T. broussonetii possesses important antimicrobial and antioxidant properties and can, therefore, be used as a natural preservative ingredient in the cosmetic industry.

Antibacterial activity of Macrosciadium alatum (M.Bieb.) plant extract.

OBJECTIVES: The flora of Azerbaijan is represented by one species of the Macrosciadium genus: Macrosciadium alatum, belonging to the Apiaceae family. It is commonly found in the Greater and Lesser Caucasus regions of Azerbaijan, as part of subalpine meadow plant communities. M. alatum is characterized by its robust, thick, tuberous roots, long-petioled and several times pinnately divided leaves, numerous (30-50) white umbels, and oval-shaped fruits. The primary objective of this research is to determine the antimicrobial potential of the aqueous extract obtained from M. alatum against both Gram-negative and Gram-positive bacteria. The plant preparations utilized in in vitro experiments were in the form of maceration, infusion, and hydrodistillation as aqueous extracts. M. alatum extract exhibited maximum (measuring 22.3 ± 1.4 mm) inhibition zones against bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Salmonella enteritidis) strains. Following exposure to the M. alatum plant extract, a significant reduction in bacterial cell cytoplasmic pH was observed (p≤0.04). METHODS: In order to investigate the antimicrobial effects of the plant extract, commonly accepted procedures were followed using well-known bacterial strains, including S. aureus, B. cereus, E. coli, S. enteritis and P. aeruginosa, which are principal causative agents of purulent-inflammatory processes. The 20 % aqueous extract was used. RESULTS: The conducted experiment to determine the impact of the plant extract on microorganisms revealed that the extract significantly affects the bacterial cell membrane. Specifically, there is a decrease in pH, and hyperpolarization of the cell membrane occurs. The efficacy of the preservative effect is highly dependent on the environmental pH. 1. The 20 % aqueous extract from exhibited antimicrobial activity and effectively preventing the development of foodborne pathogens and putrefactive microorganisms. 2. A 20 % aqueous extract of M. alatum exhibits antimicrobial activity, effectively inhibiting the growth of foodborne pathogens and spoilage microorganisms. 3. Extract led to an increase in H+ concentration within bacterial cell cytoplasm, surpassing the OH- concentration. 4. M. alatum species has a significant inhibitory effect on the growth of microorganisms such as S. aureus, E. coli, P. aeruginosa, and S. enteritidis. CONCLUSIONS: The results suggest that the extract from M. alatum possesses antimicrobial properties, making it a potential candidate for use as a natural food preservative. The observed hyperpolarization of the cell membrane and pH reduction further support its potential as an effective antibacterial agent.

Validation of a novel alcohol-free preservative solution in comparison with conventional prefixation on quality of serous body fluid smears.

BACKGROUND: We describe a novel alcohol-free preservative composed of glucose, mannitol, disodium hydrogen orthophosphate, thymol, and distilled water (glucose-mannitol-disodium dihydrogen orhtophosphate-thymol [GMDT] preservative) in appropriate proportion as an alternative to alcohol prefixation (APF) of body fluids. OBJECTIVES: To assess the cytomorphologic preservation and staining quality of serous body fluid smears generated by GMDT preservative and compare it with smears processed by standard 50% APF. METHODOLOGY: The study comprised 151 effusion samples. Each sample was equally divided into four tubes. Equal volumes of APF and GMDT preservatives were added to the first two tubes and left at room temperature for 24 h. Similarly, the corresponding preservatives were added to the third and fourth tubes and stored for 48 h. Two smears were prepared from the centrifuged sediments of each tube (all four tubes) and stained with May-Grünwald Giemsa and Papanicolaou (Pap) stains. Using a three-tiered scoring system, the smear examination was blinded to assess the extent of cellular preservation and the staining quality by two cytotechnologists and two cytopathologists. Statistical analysis was performed by STATA 16.0. RESULTS: Samples processed with the GMDT preservative at 24 h showed better cytoplasmic preservation and smear background, while nuclear features and staining quality showed no difference between the two preservatives. Mild cytoplasmic and nuclear degenerative changes were noted with the GMDT at 48 h, while all four parameters remained similar with APF at 24 and 48 h. CONCLUSIONS: The newly developed alcohol-free, GMDT preservative, could be a feasible and cost-effective alternative to 50% APF, preferably when samples are processed within 24 h.

Effect of Allium Jesdianum's extract on the physicochemical, antioxidant, antimicrobial and sensory properties of Sausage characteristics.

The study investigated the use of Allium Jesdianum plant extract as a natural preservative in sausage dough at varying concentrations. After preparation, chemical and microbial tests were conducted on the samples at zero, 14, 28, and 42 days. The study found no significant changes in pH, moisture, fat, or protein content, but the control samples consistently had the highest total volatile nitrogen (TVN) levels. The peroxide test revealed a significant difference between the control and extract samples. Sensory analysis indicated a significant difference between the control and the 200 and 300 ppm extracts (P < 0.05). Addition of Jesdianum extract significantly reduced the total viable count and psychrophilic bacteria compared to the control, subsequently extending the shelf-life of the product to over 42 days. Overall, Allium Jesdianum extract, with its antioxidant and antimicrobial properties, is beneficial in preserving sausage products and can be recommended as a nitrite substitute.

Bio-Preservation Potential and Antimicrobial Activity of Bacteriocin-Producing Lactic Acid Bacteria Isolated from Ethiopian Traditional Fermented Dairy Products.

This study aimed to assess the antibacterial activity and bio-preservation capability of bacteriocin-producing LAB isolated from Ethiopian traditional fermented dairy products in raw milk from Jimma town. Bacteriocin-producing LAB were tested for their antimicrobial activity against various foodborne pathogens, including Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, and Salmonella typhimurium. The results showed that probiotic LAB isolates inhibited foodborne pathogens (E. coli, S. aureus, and L. monocytogenes), with inhibition zones ranging from 22.00 ± 0.57 to 34.13 ± 0.57. Enterococcus faecium and Lactococcus lactis demonstrated possible antagonistic effects against E. coli, while Pediococcus pentosaceus had a 34.13 ± 0.57 mm inhibitory zone against Pseudomonas aeruginosa. The isolates also showed co-aggregation potential with the pathogens, with Lactococcus lactis isolates and their combinations demonstrating the best co-aggregation capabilities against the investigated pathogens. The bio-preservative assay showed that putative probiotic isolates (L. lactis JULABE35, E. faecium JULABE 23, and P. pentosaceus JULABE05) were efficient in decreasing Listeria monocytogenes in raw milk. After 7-8 days, milk samples diagnosed with these isolates showed complete reduction of Listeria monocytogenes. The bio-preservation capability of bacteriocin-producing LAB on raw milk extended the shelf life of milk at 4 °C storage for ten days, compared to six days for milk samples without probiotic LAB. The milk samples preserved with probiotic and bacteriocin-producing isolates showed good proximate analysis, showing significant variation with milk kept without bacteriocin-producing isolates. The isolated chemicals employed in this study can be used as food additives or food preservatives, indicating potential applications in Ethiopian traditional fermented dairy products.

Successful management of dry eye disease with a new eye drop formulation combining hyaluronic acid, trehalose, and N-acetyl-aspartyl-glutamic acid (NAAGA).

PURPOSE: To assess the clinical efficacy and safety of T2769, a new preservative-free eye drop combining hyaluronic acid (HA), trehalose and N-acetyl-aspartyl-glutamic acid (NAAGA), in dry eye patients. PATIENTS AND METHODS: This was a multicenter, non-comparative, open-label study. After a run-in period with NaCl 0.9% solution, 62 patients with moderate-to-severe dry eye disease (DED) were included and treated with T2769, 3 to 6 times/day for 42 days. The primary efficacy endpoint was the change in global ocular symptomatology assessed on a visual analog scale between Day 1 (D1) and D42. Other efficacy endpoints included ocular surface disease index (OSDI), soothing sensation, individual dry eye symptoms, conjunctival hyperemia, global ocular staining, tear break-up time, Schirmer test, and global efficacy assessed by the investigator. Safety was assessed throughout the study. RESULTS: A marked reduction in ocular symptomatology was observed from D1 to D42 with a mean change of -55.9±23.1mm (P<0.001). This was accompanied by a mean change in OSDI score from baseline of -44.6±15.9 (P<0.001) and a substantial soothing sensation in 82.3% of patients. Clinically significant improvements were observed for all ocular symptoms (mainly burning/irritation, stinging/pain, feeling of ocular dryness, foreign body sensation, itching/pruritus). Conjunctival hyperemia significantly decreased in 96.8% of patients (56.5% displayed no conjunctival hyperemia at D42 versus 0% at baseline). All signs and symptoms improved by D14 and further improved at D42. Investigators assessed the ocular efficacy of T2769 as very satisfactory or satisfactory for 91.9% of patients at D42. T2769 was well tolerated, with no ocular adverse events and only a few ocular symptoms upon instillation. CONCLUSION: Management of moderate-to-severe DED patients with the new formulation T2769, combining NAAGA to HA and trehalose, led to rapid and significant improvements in dry eye signs and symptoms with good tolerability.

24-Hour Evaluation of the Effectiveness and Tolerability of Preservative-Free Tafluprost-Timolol Fixed Combination in Open-Angle Glaucoma or Ocular Hypertensive Patients Previously Treated with Preserved Latanoprost.

Purpose: The effectiveness, safety and tolerability of the preservative-free fixed combination of tafluprost and timolol (PF-TTFC) were evaluated over the 24-h in patients with open-angle glaucoma or ocular hypertension showing signs and symptoms of Ocular Surface Disease (OSD) and uncontrolled intraocular pressure (IOP) on prior benzalkonium chloride (BAK) - Latanoprost monotherapy. Methods: In this multi-center, prospective, interventional, non-comparative clinical trial, patients treated with BAK-Latanoprost underwent 24-h IOP measurements (8 am, 11 am, 2 pm, 5 pm, 8 pm, 11 pm, 2 am, 5 am) at baseline and after 3 months from switch to PF-TTFC. Mean 24-h IOP and daytime (8 am-8 pm) vs nighttime (11 pm - 5 am) IOP were compared. Changes in OSD signs and symptoms, quality of life (QoL) and in-vivo corneal confocal microscopy (IVCM) were also evaluated. Results: Thirty-eight patients were analyzed. The mean 24-h IOP significantly decreased after 3 months from 17.8 mmHg (95% CI: 17.1-18.6) to 15.3 mmHg (95% CI: 14.6-16.1, p < 0.001). IOP was significantly reduced both at daytime (p < 0.001) and nighttime (p < 0.001), with better IOP control at night [-2.9 (95% CI: -3.5 to -2.1) mmHg vs -2.3 (95% CI: -2.9 to -1.6) mmHg]. In 20 patients (52.6%), corneal fluorescein staining improved, whereas in 4 patients (10.5%) it worsened. Hyperemia has improved in 24 (63.3%) patients and worsened in 2 (5.3%). Breakup time, Schirmer test and QoL scores showed no changes. At IVCM, the mean corneal wing-cell size was found significantly decreased (p < 0.005). Conclusion: The switch from BAK-Latanoprost to PF-TTFC significantly reduced IOP over the 24-h and improved OSD signs and symptoms.

Heterologous expression of frog antimicrobial peptide Odorranain-C1 in Pichia pastoris: Biological characteristics and its application in food preservation.

To reduce food spoilage and deterioration caused by microbial contamination, antimicrobial peptides (AMPs) have gradually gained attention as a biological preservative. Odorranain-C1 is an α-helical cationic antimicrobial peptide extracted from the skin of frogs with broad-spectrum antimicrobial activity. In this study, we achieved the expression of Odorranain-C1 in Pichia pastoris (P. pastoris) (also known as Komagataella phaffii) by employing DNA recombination technology. The recombinant Odorranain-C1 showed broad-spectrum antibacterial activity and displayed a minimum inhibitory concentration within the range of 8-12 µg.mL-1. Meanwhile, Odorranain-C1 exhibited superior stability and lower hemolytic activity. Mechanistically, Odorranain-C1 disrupted the bacterial membrane's integrity, ultimately causing membrane rupture and subsequent cell death. In tilapia fillets preservation, Odorranain-C1 inhibited the total colony growth and pH variations, while also reducing the production of total volatile basic nitrogen (TVB-N) and thiobarbituric acid (TBA). In conclusion, these studies demonstrated the efficient recombinant expression of Odorranain-C1 in P. pastoris, highlighting its promising utilization in food preservation.

Determination of Post-Fermentation Waste from Fermented Vegetables as Potential Substitutes for Preservatives in o/w Emulsion.

Post-fermentation wastes are rich sources of various biologically active compounds with antimicrobial activity, whose potential is not being fully exploited. One of the possible applications of post-fermentation waste may be its use as a natural preservative that effectively combats pathogens found in formulations. The study aims included the following: (1) compare the antimicrobial and antioxidant activity of fermented vegetable extracts (FVEs), (2) examine the inhibition of cosmetic-borne pathogens by FVEs, and (3) estimate the preservative effectiveness of FVEs in o/w emulsions. It was found that fermented white cabbage, cucumber, celery, and the mixture of fermented white cabbage, cucumber, and celery (1:1:1) showed antibacterial and antifungal activity against all the tested reference microbial strains. The addition of fermented cucumber, celery, and the mixture of fermented white cabbage, cucumber, and celery (1:1:1) to the o/w emulsion fulfilled criterion A of the preservative effectiveness test for S. aureus, E. coli, and A. brasiliensis, but did not fulfill the criterion for P. aeruginosa and C. albicans. The tested FVEs have comparable activity to inhibit pathogens in o/w emulsion as sodium benzoate. The results of our study prove that FVEs can be valuable raw materials supporting the preservative system, which, in turn, can significantly reduce the concentration of preservatives used in o/w emulsion.

Comparison of multiple preservation and digestion methods for determination of tungsten concentrations in environmental media using ICP-MS.

Tungsten is an emerging environmental pollutant. However, a proved robust method for preserving and determining the concentrations of tungsten in environmental media is still lacking. This study examined and compared the suitability of classic methods and previously reported tungsten-oriented methods on preserving dissolved tungsten and recovering tungsten from soil/sediment matrix. Tungsten concentrations in the water samples and digestates were then determined by inductively coupled plasma mass spectrometry. Our data showed that the tungsten-oriented HF and alkaline preservatives indeed successfully maintained the stability of dissolved tungsten. Even when preserved using HNO3 or HCl, dissolved tungsten concentrations did not notably change in most of our water samples over the course of ∼4 months. Using glass containers for storing water samples also did not produce much difference from using high-density polyethylene containers. Our data further suggested that the addition of HF in digestion was important for tungsten solubilization from soil/sediment matrix. The digestion methods with HNO3/HCl/HF and HNO3/HF/NH4OH/EDTA both yielded quantitative recoveries of tungsten from certified reference materials and known synthetic samples, while the other tested methods had limited recoveries. The methods validated by this study could be used to accurately determine tungsten concentrations in environmental media and thereby to assess the fate and potential risks of tungsten.

Unveiling the Future of Meat Packaging: Functional Biodegradable Packaging Preserving Meat Quality and Safety.

Meat quality and shelf life are important parameters affecting consumer perception and safety. Several factors contribute to the deterioration and spoilage of meat products, including microbial growth, chemical reactions in the food's constituents, protein denaturation, lipid oxidation, and discoloration. This study reviewed the development of functional packaging biomaterials that interact with food and the environment to improve food's sensory properties and consumer safety. Bioactive packaging incorporates additive compounds such as essential oils, natural extracts, and chemical substances to produce composite polymers and polymer blends. The findings showed that the incorporation of additive compounds enhanced the packaging's functionality and improved the compatibility of the polymer-polymer matrices and that between the polymers and active compounds. Food preservatives are alternative substances for food packaging that prevent food spoilage and preserve quality. The safety of food contact materials, especially the flavor/odor contamination from the packaging to the food and the mass transfer from the food to the packaging, was also assessed. Flavor is a key factor in consumer purchasing decisions and also determines the quality and safety of meat products. Novel functional packaging can be used to preserve the quality and safety of packaged meat products.

Natural approach of using nisin and its nanoform as food bio-preservatives against methicillin resistant Staphylococcus aureus and E.coli O157:H7 in yoghurt.

BACKGROUND: Natural antimicrobial agents such as nisin were used to control the growth of foodborne pathogens in dairy products. The current study aimed to examine the inhibitory effect of pure nisin and nisin nanoparticles (nisin NPs) against methicillin resistant Staphylococcus aureus (MRSA) and E.coli O157:H7 during the manufacturing and storage of yoghurt. Nisin NPs were prepared using new, natural, and safe nano-precipitation method by acetic acid. The prepared NPs were characterized using zeta-sizer and transmission electron microscopy (TEM). In addition, the cytotoxicity of nisin NPs on vero cells was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The minimum inhibitory concentrations (MICs) of nisin and its nanoparticles were determined using agar well-diffusion method. Further, fresh buffalo's milk was inoculated with MRSA or E.coli O157:H7 (1 × 106 CFU/ml) with the addition of either nisin or nisin NPs, and then the inoculated milk was used for yoghurt making. The organoleptic properties, pH and bacterial load of the obtained yoghurt were evaluated during storage in comparison to control group. RESULTS: The obtained results showed a strong antibacterial activity of nisin NPs (0.125 mg/mL) against MRSA and E.coli O157:H7 in comparison with control and pure nisin groups. Notably, complete eradication of MRSA and E.coli O157:H7 was observed in yoghurt formulated with nisin NPs after 24 h and 5th day of storage, respectively. The shelf life of yoghurt inoculated with nisin nanoparticles was extended than those manufactured without addition of such nanoparticles. CONCLUSIONS: Overall, the present study indicated that the addition of nisin NPs during processing of yoghurt could be a useful tool for food preservation against MRSA and E.coli O157:H7 in dairy industry.

Comparative study on effect of pomegranate peel powder as natural preservative and chemical preservatives on quality and shelf life of muffins.

This research aims to investigate the potential of utilizing pomegranate peel powder (PPP) as a natural preservative in muffin preparation. Pomegranate peel is a rich source of bioactive compounds, including phenolics, flavonoids, and tannins, which possess high antioxidant and antimicrobial properties. The In-Vitro antifungal activity of pomegranate peel powder (8% PPP), potassium sorbate (0.1% PS) and calcium propionate (0.5% CP) was assessed against Penicillium sp. and Aspergillus sp. using poison food technique. The PPP showed the anti-fungal activity by delaying the growth of microorganism on media plate similar to the PS and CP. The effect of utilization of PPP on quality characteristics of muffins were compared with the muffins with chemical preservatives (0.1% PS and 0.5% CP). The viscosity and specific gravity of batter significantly increased from 7.98 to 11.87 Pa s and 1.089-1.398 respectively on addition of 8% PPP. The optical microscopic structure of PPP added batter revealed the decrease in the number of air cells from 24 to 12 with radius range of 6.42-72.72 µm and area range of 511.03-15,383.17 µm2. The functional properties of flour with PPP had higher water absorption capacity, foaming stability, emulsification activity and emulsion stability than others. The addition of PPP significantly increase the weight (32.83 g), and decrease the height (31.3 mm), volume (61.43 cm3), specific volume (1.67 cm3/g) and baking loss (10.19%). The 418.36% increase in fibre content, 14.46% and 18.46% decrease in carbohydrates and energy value was observed in muffin with 8% PPP as compared to control respectively. The total phenols was increased from 0.92 to 12.5 mg GAE/100 g, total tannin from 0.2 to 8.27 mg GAE/100 g, In-vitro antioxidant activity by DPPH from 6.97 to 29.34% and In-vitro antioxidant activity by FRAP from 0.497 to 2.934 mg AAE/100 g in muffins added with 8% PPP. The muffin with PPP was softer than control and muffin with 0.1% PS. The addition of PPP resulted to improve in muffin texture but taste slightly bitter. During the storage of muffins at room temperature (27-30 °C), the moisture content of muffin with PPP was reduced from 17.04 to 13.23% which was higher than the rest of the treatments. Similarly, the hardness of sample with PPP was higher than the sample with 0.5% CP, but lowers than control and sample with 0.1% PS throughout the storage period. The results suggest that pomegranate peel powder can be successfully used as a natural preservative in place of chemical preservatives in muffins, to extend the shelf life. This study provides the opportunity to use PPP as functional ingredient and natural preservative in different bakery products.

Integrated high-throughput drug screening microfluidic system for comprehensive ocular toxicity assessment.

Traditional experimental methodologies suffer from a few limitations in the toxicological evaluation of the preservatives added to eye drops. In this study, we overcame these limitations by using a microfluidic device. We developed a microfluidic system featuring a gradient concentration generator for preservative dosage control with microvalves and micropumps, automatically regulated by a programmable Arduino board. This system facilitated the simultaneous toxicological evaluation of human corneal epithelial cells against eight different concentrations of preservatives, allowing for quadruplicate experiments in a single run. In our study, the IC50 values for healthy eyes and those affected with dry eyes syndrome showed an approximately twofold difference. This variation is likely attributable to the duration for which the preservative remained in contact with corneal cells before being washed off by the medium, suggesting the significance of exposure time in the cytotoxic effect of preservatives. Our microfluidic system, automated by Arduino, simulated healthy and dry eye environments to study benzalkonium chloride toxicity and revealed significant differences in cell viability, with IC50 values of 0.0033% for healthy eyes and 0.0017% for dry eyes. In summary, we implemented the pinch-to-zoom feature of an electronic tablet in our microfluidic system, offering innovative alternatives for eye research.

High-throughput DNA extraction strategy for fecal microbiome studies.

Microbiome studies are becoming larger in size to detect the potentially small effect that environmental factors have on our gut microbiomes, or that the microbiome has on our health. Therefore, fast and reproducible DNA isolation methods are needed to handle thousands of fecal samples. We used the Chemagic 360 chemistry and Magnetic Separation Module I (MSMI) instrument to compare two sample preservatives and four different pre-treatment protocols to find an optimal method for DNA isolation from thousands of fecal samples. The pre-treatments included bead beating, sample handling in tube and plate format, and proteinase K incubation. The optimal method offers a sufficient yield of high-quality DNA without contamination. Three human fecal samples (adult, senior, and infant) with technical replicates were extracted. The extraction included negative controls (OMNIgeneGUT, DNA/RNA shield fluid, and Chemagic Lysis Buffer 1) to detect cross-contamination and ZymoBIOMICS Gut Microbiome Standard as a positive control to mimic the human gut microbiome and assess sensitivity of the extraction method. All samples were extracted using Chemagic DNA Stool 200 H96 kit (PerkinElmer, Finland). The samples were collected in two preservatives, OMNIgeneGUT and DNA/RNA shield fluid. DNA quantity was measured using Qubit-fluorometer, DNA purity and quality using gel electrophoresis, and taxonomic signatures with 16S rRNA gene-based sequencing with V3V4 and V4 regions. Bead beating increased bacterial diversity. The largest increase was detected in gram-positive genera Blautia, Bifidobacterium, and Ruminococcus. Preservatives showed minor differences in bacterial abundances. The profiles between the V3V4 and V4 regions differed considerably with lower diversity samples. Negative controls showed signs from genera abundant in fecal samples. Technical replicates of the Gut Standard and stool samples showed low variation. The selected isolation protocol included recommended steps from manufacturer as well as bead beating. Bead beating was found to be necessary to detect hard-to-lyse bacteria. The protocol was reproducible in terms of DNA yield among different stool replicates and the ZymoBIOMICS Gut Microbiome Standard. The MSM1 instrument and pre-treatment in a 96-format offered the possibility of automation and handling of large sample collections. Both preservatives were feasible in terms of sample handling and had low variation in taxonomic signatures. The 16S rRNA target region had a high impact on the composition of the bacterial profile. IMPORTANCE: Next-generation sequencing (NGS) is a widely used method for determining the composition of the gut microbiota. Due to the differences in the gut microbiota composition between individuals, microbiome studies have expanded into large population studies to maximize detection of small effects on microbe-host interactions. Thus, the demand for a rapid and reliable microbial profiling is continuously increasing, making the optimization of high-throughput 96-format DNA extraction integral for NGS-based downstream applications. However, experimental protocols are prone to bias and errors from sample collection and storage, to DNA extraction, primer selection and sequencing, and bioinformatics analyses. Methodological bias can contribute to differences in microbiome profiles, causing variability across studies and laboratories using different protocols. To improve consistency and confidence of the measurements, the standardization of microbiome analysis methods has been recognized in many fields.

Glaucoma Drug Prescription Pattern in North India: Public vs Private Sector Hospitals.

Background: Glaucoma is an optic neuropathy associated with characteristic structural damage to the optic nerve and associated visual dysfunction that may be caused by various pathological processes. A number of pharmacological agents are used to reduce the intraocular pressure (IOP), involving the usage of two or three medications concurrently. Literature is sparse regarding prescription patterns of antiglaucoma drugs, especially regarding variability in public sector vs private sector hospitals. Drug utilization studies can add insight for crafting rational, affordable, and ocular surface friendly prescriptions. Aim: This study assessed the prescription pattern in glaucoma patients of a public sector, tertiary care hospital vs a private sector tertiary care hospital. Materials and methods: In this retrospective study, pertinent data of diagnosed and labeled glaucoma patients were reviewed. Data collected included demographic details, type of glaucoma, number and nature of drugs prescribed, whether innovator or generic drugs were prescribed, if fixed-drug combinations (FDCs) and preservative-free formulations were prescribed. The prescription patterns between the two sectors were compared, as were the prescription patterns between primary open-angle glaucoma (POAG) and primary angle-closure disease (PACD). Results: A total of 336 prescriptions were evaluated (216 from public sector, group I; 120 from private sector, group II). Travoprost 0.004% was the most prescribed antiglaucoma medication in both group I (30.09%) and group II (38.33%). Brimonidine and brinzolamide (14.17%) was the most prescribed combination in group II, while Brimonidine with Timolol (7.87%) in group I. In group I, Timolol and Travoprost were the most prescribed medications for both PACD and POAG. Conclusion: This study showed that both public sector as well as private sector tertiary care centers prescribe antiglaucoma medications in tune with current principles of rational drug use. Preservative-free drugs were preferred in both the groups for better adherence. How to cite this article: Bhartiya S, Ichhpujani P, Parmar UPS, et al.Glaucoma Drug Prescription Pattern in North India: Public vs Private Sector Hospitals. J Curr Glaucoma Pract 2024;18(1):16-22.