TM by NanoLayr Ltd., containing poly(methyl methacrylate)/ethylene vinyl alcohol nanofibres. The outstanding uniformity of the nanofibres was indicated by the good correlation between pressure drop (ΔP) and areal weight with R2 values in the range of 0.82 to 0.98 across various test air velocities. By adjusting the nanofibre areal weight (basis weight), the nanofibre filter media was shown to meet the particle filtration efficiency and breathability requirements of the following internationally accepted facemask and respirator standards: N95 respirator facemask performance in accordance with NIOSH 42CFR84 , Level 2 surgical facemask performance in accordance with ASTM F2299 (filtration efficiency of up to 99.97% at 100 nm particle size and a pressure drop of 44 Pa at 8 L·min−1), and Level 2 filtration efficiency and Level 1 breathability for barrier face coverings in accordance with ASTM F3502 (filtration efficiency of up to 99.68% and a pressure drop of 133 Pa at 60 L·min−1), with Level 2 breathability being achievable at lower nanofibre areal weights.Electrospun nanofibres can outperform their melt-blown counterparts in many applications, especially air filtration. The different filtration mechanisms of nanofibres are particularly important when it comes to the air filtration of viruses (such as COVID-19) and bacteria. In this work, we present an electrospun nanofibre filter media, FilterLayr Filtration plays an important role in purifying and decontaminating two of life’s necessities: water and air. Increasing awareness of the links between air quality and human health has led to a rise in the demand for improved personal protection from airborne pollutants and disease-causing microbes. In this respect, electrospun nanofibres (NFs) provide unique capabilities when used as an active protective layer in facemasks. When compared with commonly used filters containing melt-blown (MB) fibres, electrospun NFs provide enhanced protection against airborne particles, bacteria, and viruses such as COVID-19 ,3,4,5,6.Electrospinning (ES) is considered the most suitable method for producing polymeric NFs. This is due to the versatility of ES and its ability to use a wide variety of polymers at both lab and production scale . HistoriThe ES process can be used to generate functional NFs, which can provide enhanced properties as well as higher surface areas when compared with MB fibres. The high surface areas of ES NFs make them particularly suitable for functionalisation for a wide range of applications including filter media, catalysis, super absorbents, scaffolds for tissue engineering and wound dressings, energy storage, and electronics . The ES ES uses electrostatic forces to draw charged threads from a polymer solution to create NFs. The diameters of these NFs typically range from 10 to 600 nanometres (nm) . The fibNF filter media produced by ES has attracted much interest in air filtration applications. This is partly due to the fact that the diameters of NFs are 10–100 times smaller than those of conventional MB microfibres . NFs areTo understand how ES NFs enhance filtration performance, it is important to understand the particle-capturing mechanisms. Particles can be blocked by a filter via five different mechanisms: Sieving, Interception, Inertia Impaction, Diffusion, and Electrostatic Attraction . GravityDue to the various mechanisms by which filtration occurs, the smallest particles are typically not the most difficult to filter out. Most air filters have a region of lower filtration efficiency somewhere between 0.1–0.5 µm . ParticlAs discussed above, some particles in the nano range (100~500 nm) are difficult to filter as they do not conform to one particular capture mechanism. Filtration of MPPS particles require multiple and uniform layers of NF, which defer the particles so that they obey one of the capture mechanisms. Multilayer filters are often hindered by poor breathability and high pressure drop, which is undesirable for air filtration. However, electrospinning enables control of the porosity, packing density, fibre diameter, and surface area of the NFs. Modification of these parameters allows for customisation of NF air filters to allow for the filtration of a wider range of particles/contaminant types, sizes and concentrations can then be achieved with MB filters. As shown in The recent COVID-19 pandemic is caused by the novel coronavirus, SARS-CoV-2, which is transmitted primarily through direct contact with respiratory droplets of an infected person (generated through coughing and sneezing) ,21. The Various ES solutions were prepared using the following materials: formic acid, acetic acid, ethylene vinyl alcohol (EVOH), and poly(methyl methacrylate) (PMMA) supplied by Merck; spunbonded polypropylene nonwoven substrate fabric (SPP).2 samples on a Precisa XB220A analytical balance and then dividing the sample mass by the sample area. The unit of the areal weight is gram per square metre (GSM).Electrospinning is the most commonly used technique for the mass-production of polymer NFs and has been explained in detail by Rutledge and Fridrikh . A uniquNF samples were analysed using a JEOL JCM-5000 scanning electron microscope  and fibre diameters were measured using an evaluation software called Fibraquant, whereby the average NF diameters were determined with from 50 to 100 measurement readings. Pressure drop is the measure of the difference in total pressure between two different points of a fluid (air) as it flows through the filter media. This pressure drop is due to frictional forces caused by the resistance to flow. The pressure drop of different filter media, including samples containing NF at different areal weights, were measured using a TexTest FX 3300 LabAir IV  in accordance with ASTM Test Method F2299.Breathing resistance or breathability is a measure of the difficulty to breathe in or out through a mask or filter media and is commonly expressed as a pressure drop across the fabric layers. Different filter media including NFs at different areal weights were measured using a PALAS PMFT 1000  in accordance with ASTM Test Method D3502.filter = Cdown/Cup × 100%). PFE is the complement of filter penetration (PFE (%) = 100% − Pfilter). A filter media with a PFE of 98% will block 98% of particles  so that only 2% of particles will pass through the filter media when air is inhaled or exhaled. It should be noted that the values for PFE presented in this paper are measured only on the filter media and not on fabricated masks. The PFE of facemasks may vary during filter testing due to air leakage around the edge of the mask, and it is typically the responsibility of the mask manufacturer to guarantee compliance by further testing of their final product. Different filter media including those containing NFs at various areal weights were measured using a PALAS PMFT 1000 according to NIOSH 42CFR84 (N95), ASTM Test Method D3502, and ASTM Test Method F2299. A comparative summary of the filtration test methods of all three of these international test standards can be seen in Particle filtration efficiency (PFE) is a measure of the proportion of particles that are intercepted by the mask or filter material. The general approach to determine the PFE is to challenge the test sample with small particles that are carried in the air and move through the test specimen at a specific airflow velocity , and to measure the particle concentration upstream (before) and downstream (after) of the test sample. The ratio between the particle concentration downstream and the particle concentration upstream is the filter penetration (P3 at 23 °C. Electrospinning solutions were prepared by adding PMMA and EVOH granules (15 wt%) slowly into a blend of formic acid (99% purity) and glacial acetic acid and stirred at room temperature until all the particles had fully dissolved. The PMMA used in this research has an average molecular weight of 120,000, a glass transition temperature of 105 °C, and density of 1.188 g/mL at 25 °C. The EVOH used contains 38 mol% ethylene, has a glass transition temperature of 58 °C, an MFR of 8.0 g/10 min, and a density of 1.17 g/cmTM, is a 3-layer structure consisting of two outer layers of nonwoven fabric with a middle layer consisting of kilometre-long NFs  increased when the NF areal weight increased from 0.65 gsm to 2.1 gsm. All the samples tested showed a linear relationship between pressure drop and areal weight, with R2 values ranging from 0.82 to 0.98 to verify the uniformity of the NF layer in the filter media.By combining the NF layer with a stronger polypropylene substrate, the presented material has sufficient mechanical strength to be used in filter applications. It has been shown that many commercially available NF filters often have lower filtration efficiencies than what is reported in their design specifications due to the nonuniformity of the NF, especially when the NFs are produced using conventional needle-based electrospinning techniques. This is mostly because the electric field between the conductive needle tip and the grounded substrate surface is nonuniform due to the uneven nature of the substrate fabric ,38,39,40−1, as required for the PFE test method in ASTM F2299. All filter media samples containing NF layers of more than 1.1 gsm met the particulate filtration requirement for Level 2 ASTM F2299 facemasks and showed filtration efficiencies of ≥98% (−1). The detailed ASTM F2299 pressure drop and filtration efficiency results at particle sizes of 100 nm, 300 nm, and 500 nm are displayed in −1 in accordance with the NIOSH 42CFR84 test method. In this case, all filter media samples containing NF layers of more than 1.5 gsm met the N95 facemask filtration efficiency requirements of NIOSH 42CFR84, and all samples showed pressure drops below 330 Pa at 120 L·min−1. To pass the standard requirements of both filtration efficiency and breathability, the desired areal weight ranges from 1.50 to 1.63 gsm. For the standard specification for barrier face coverings, ASTM F3502, a minimum filtration efficiency of 20% is necessary to meet the requirements of level 1, and ≥50% filtration efficiency is required to meet the requirements of level 2  containing electrospun PMMA/EVOH NFs has been developed by NanoLayr Ltd. NF filters are excellent for use in air filtration applications, since they can achieve enhanced particle capture while offering low pressure drop and homogeneous porosity. In this study, filter media with NF areal weights ranging from 0.65 gsm to 2.1 gsm were analysed using a PALAS PMFT 1000 Filter Test System and a TexTest FX 3300 LabAir IV Air Permeability Tester.A novel air filtration media , and ASTM F2299 Level 2 was achieved for samples with NF areal weights exceeding 1 gsm (with a filtration efficiency reaching 99.97% for 300 nm-sized particles and a pressure drop of 44 Pa). All tested samples outperformed the minimum requirements of ASTM F3502 for Level 2 filtration efficiency and Level 1 breathability (with a filtration efficiency of up to 99.68% and a pressure drop below 133 Pa), with Level 2 breathability being achievable only at lower NF areal weights. It was also clear from the results that better filtration efficiencies could be achieved with heavier NF layers, and this was more prevalent with the filtration of smaller aerosol particle sizes. It can be concluded that NanoLayr Ltd. achieved the industrial-scale production of a novel nanofibre filter media, which, unlike other nanofibre media, can meet the filtration efficiency and pressure drop requirements of three major and significantly different international test standards for respirator facemasks, surgical facemasks, and barrier face coverings. There is, however, an increasing need for the development of self-decontaminating facemask and filter media that have the added benefits of being able to inactivate the pathogens, viruses, and bacteria that they capture [


Zea mays L. is one of the most produced crops, and there are still parts of the world where maize is the basic staple food. To improve agriculture, mankind always looks for new, better methods of growing crops, especially in the current changing climatic conditions. Cold atmospheric pressure plasma (CAPP) has already showed its potential to enhance the culturing of crops, but it still needs more research for safe implementation into agriculture. In this work, it was shown that short CAPP treatment of maize grains had a positive effect on the vitality of grains and young seedlings, which may be connected to stimulation of antioxidant and lytic enzyme activities by short CAPP treatment. However, the prolonged treatment had a negative impact on the germination, growth, and production indexes. CAPP treatment caused the increased expression of genes for heat shock proteins HSP101 and HSP70 in the first two days after sowing. Using comet assay it was observed that shorter treatment times (30–120 s) did not cause DNA damage. Surface diagnostics of plasma-treated grains showed that plasma increases the hydrophilicity of the surface but does not damage the chemical bonds on the surface. It belongs to the most produced cereal crops, together with wheat and rice, and its production is estimated to grow steadily in the next decade. It is an important staple food in many parts of the world, and maize for human consumption is estimated to rise, especially in Sub-Saharan Africa. Besides food production for humans, there is a strong demand for bioethanol and animal feed and other maize products such as corn starch or syrup [Maize or syrup . It is t2-containing gases such as air), and high-energy electrons. Cold atmospheric pressure plasma (CAPP) is very suitable for practical use. Its production does not require a low-pressure vacuum device. The exact composition of non-thermal plasma (NTP) depends on the type of plasma source and the working gas used. For example, CAPP generated in different ratios of O2:N2 by Diffuse Coplanar Surface Barrier Discharge (DCSBD) analyzed by Fourier Transform Infrared spectroscopy (FTIR) and optical emission spectroscopy showed the presence of oxygen and nitrogen species  and excited molecules  responsible for producing radiation in the UV region by deexcitation. Additionally, a relatively high amount of ozone is produced in an oxygen-rich mixture [Plasma is ionized gas containing various reactive, neutral, and charged particles. Non-thermal, low-temperature or “cold plasma” means that heavy particles—ions and neutrals—remain near room temperature, and electrons reach high temperature due to their high mobility ,3,4. TheThe use of non-thermal plasma in agriculture is a promising concept since many studies state that plasma treatment of seeds can enhance germination and growth parameters of seedlings ,7,8,9 an2−, ˙OH, NO) may play a role in the breaking of dormancy and enhancing seeds’ germination [2O2 in roots and leaves of Vigna mungo L., especially for longer exposure times, after the treatment of seeds with NTP. Some authors noticed an increase in the activity of antioxidant enzymes, such as superoxide dismutase, catalase, or peroxidases [There are many reactive oxygen and nitrogen species (RONS) in the NTP. On the one hand, RONS contained in NTP , which leads to the short treatment times (up to 20–50 s), suitable for practical applications [Differences in the effects of NTP treatment between publications may be caused by the variety of plasma devices. For example, DCSBD plasma source is CAPP, specified by the high surface and volume power density  are among the first to respond. As molecular chaperones, their role is to refold, disaggregate, or send to degradation misfolded proteins and prevent aggregation. The activity of HSPs and heat shock factors (HSFs) is also important in priming or acquired tolerance to stress ,32. For g et al. . HSFA4A bidopsis . HSP70s y metals .This paper studied the influence of CAPP treatment of maize grains on the germination and growth parameters and activities of lytic and antioxidant enzymes. We verified physical and chemical changes on the maize grains surface due to plasma treatment in different working gases. We also monitored the DNA damage caused by this treatment and looked at the expression of some heat shock protein genes to find if they are involved in the response of maize to the CAPP treatment.−1 the significant peaks attributed to symmetric and asymmetric stretching of methyl and methylene (CH3 and CH2) group can be visible, originating primarily from lipids [−1 in “the fingerprint” region [−1 and other bonds in region 1200–800 cm−1 [−1, amide II (stretching of C–N and deformation of N–H) in 1550–1520 cm−1 and amide III  at about 1240 cm−1. In addition, the weak band of amide A (N–H stretching vibration) can be found in a spectra of proteins at about 3200 cm−1, but it is overlapped by a strong, wide band of OH stretching in 3600–3000 cm−1 [In Attenuated Total Reflectance-Fourier Transform Infrared spectroscopy (ATR-FTIR) spectra acquired from the surface of maize grains, we can identify the characteristic chemical groups attributed to lipids, carbohydrates, and proteins. In 3000–2800 cmm lipids ,38. Addi” region . Carbohy800 cm−1 . Three d000 cm−1 .−1, where we can observe the decrease of C–H bonds, and in interval 1800–1550 cm−1, in which the carbonyl (C=O) group is typical. This effect is considerable on samples treated by air and oxygen plasma. A decrease in chemical groups typical for lipids indicate the activation of the maize surface by oxidation and removal of the top lipid layer  measured on untreated samples was 107.9 ± 4.1°, thus, the surface is considered hydrophobic (WCA > 90°). After plasma treatment, the contact angle significantly decreased even at a short exposure time. After 30 s of treatment, the value of WCA was reduced approximately by 50% in all working gases. The value below <5°corresponds to the complete spreading out of the droplets, which was reached in air (A) and oxygen (O) plasma after 300 s of treatment. In nitrogen (N), the contact angles were higher after a longer treatment time (180 s and 300 s) . This efSeed germination is the first and very critical period in the life cycle of each plant. The seeds must absorb water to activate their metabolism, and this hydration kinetics takes place in three phases ,43. FirsStatistically, significantly more water was taken up by maize grains treated with CAPP which was generated in an oxygen atmosphere for 30, 180 and 300 s, and in CAPP which was generated in ambient air for 60 and 300 seconds A. Many aAs the maize grains  used in this study had approximately 95% germination, we did not notice an increase in the germination percentage due to CAPP application. However, the germination rate at the low exposure times  remained statistically comparable to the untreated control. On the other hand, the quicker germination of maize grains after exposure to the magnetic field might be due to increased activities of germination related enzymes, fast hydration of membranes and greater molecular mobility of water . DespiteTo find out the influence of plasma on biochemical mechanisms of plant germination, we also monitored the total soluble protein content and activities of lytic and antioxidant enzymes. The concentration of total soluble protein and the activity of lytic and antioxidant enzymes in three-day-old maize seedlings were affected by CAPP. In the case of the concentration of total soluble proteins (SP), we recorded an increase in all variants, and the trend changed depending on the working gas in which the CAPP was generated. Plasma generated in an atmosphere of oxygen and nitrogen caused an increase  in the concentration of total SP with increasing time of exposure to CAPP, compared to the untreated control. In the case of CAPP generated in ambient air, the concentration of total SP increased linearly until the treatment time of 120 s. After that there was a decrease, but even in 180 and 300 s treatment, the concentration of total SP was significantly higher (about two times higher) than in the untreated control . The incProteases are known to control signaling pathways in imbibed grains and plants. They play an important role, especially during germination, in the mobilization of storage proteins, such as free amino acids that can be used to build proteins and enzymes essential for embryo development . HoweverConcentrations of total SP in individual variants correlated with protease activity 2, N2O, NO, O3, HNO3, HNO2, CO2, ˙OH−, ˙O2−, 1O2. One of the possible mechanisms for plasma-induced stimulation of seed/grain germination is that the biochemical processes inside the seeds are activated by RONS [2O2 [Low-temperature plasma generated in ambient air usually consists of reactive oxygen and nitrogen species such as NO by RONS . Reactiv by RONS . Emergin by RONS ,66,67, b by RONS . BecauseONS [2O2 .After evaluating the activity of antioxidant enzymes, we can state that CAPP treatment did not increase oxidative stress in three-day-old maize seedlings. The activities of SOD in the treated variants were comparable to the untreated control B; the ac3) during the generation of plasma in an oxygen atmosphere, nitrogen oxides during generation of plasma in ambient air, and UV radiation during generation of plasma in a nitrogen atmosphere [Published works show that high application doses of cold plasma (180 or more seconds) negatively affect the activity of antioxidant enzymes. In pea seeds, the application of CAPP generated in the nitrogen atmosphere leads to a significant accumulation of superoxide radicals which indicates great oxidative stress . This, imosphere , which amosphere ,71,72. Omosphere ,20 and mmosphere . From thThe neutral comet assay was used to assess the DNA damage caused by CAPP treatment. Based on the germination percentage, we decided to work only with four treatment times (30–120 s) since the average germination was already significantly decreased at 120 s. We found that the pre-sowing treatment of maize grains with CAPP generated in ambient air did not cause significant damage for shorter treatment times (30–90 s) and was similar to that of negative control . There was only a slight increase (10%) in DNA damage in the 120 s variant A. For plThe DNA damage caused by CAPP treatment is usually not excessively high and increases with increasing treatment time, as is shown in other publications ,19,20,21HSP101 and HSP70 genes and heat shock factor gene HSF17 during the 72 h after sowing by qPCR in samples where dry maize grains were treated with CAPP generated in ambient air.As chaperones, heat shock proteins are essential for the cell not only in development and under normal conditions, but they are also crucial in stressful conditions, as stress often leads to various damages or denaturation of proteins . We moniHSP101 transcripts in maize shortly after the germination [HSP101 and HSP70 in the control sample at 72 h, and, on the contrary, lower expression in plasma-treated samples. A possible explanation is that it is related to the developmental processes of 72 h germination of young seedlings. Since the expression of both genes was influenced by CAPP treatment, it is possible that at 72 h, there is higher demand for chaperones in the non-treated control sample than in the CAPP-treated samples, where there was more of them in the past 48 h. Both HSP101 and HSP70 have essential roles during the heat, and other stresses [We noticed that the expression of HSP101 and HSP70 reacted to the CAPP treatment of maize grains in a similar way. Their relative expression at 24 h increased with increasing CAPP treatment time (30–120 s) compared to the non-treated control (0 s). The expression in plasma-treated samples remained higher even at 48 h compared to the non-treated control. However, a different trend occurred at 72 h, where the highest expression was in a non-treated sample. At the same time, the CAPP treatment of grains resulted in a lower relative amount of transcripts  A,B. The mination ,76. Howestresses ,78 and ostresses .Arabidopsis, HSFA2, is an integral part of the maintenance of acquired thermotolerance in Arabidopsis thaliana [A. thaliana to anoxia [Heat shock factors are regulators of HSPs expression. Therefore, we included the HSF17 gene in the analysis of HSPs expression. A homologue of HSF17 in thaliana ,34, but o anoxia . We obseo anoxia  observedZea mays L. cv. Ronaldinio) grains used in these experiments were obtained from the Central Control and Testing Institute in Agriculture in Bratislava and KWS Semená s.r.o., Bratislava, Slovakia in 2017. Grains were stored in a fridge at 8 ± 2 °C in the dark.Dried maize . The DCSBD system was supplied by sinusoidal voltage with peak-to-peak amplitude of 20 kV and frequency 15 kHz.For the plasma treatment of the maize grains, the Diffuse Coplanar Surface Barrier Discharge (DCSBD)  was used−1 during 20 scans for background/sample and resolution of 4 cm−1.Attenuated Total Reflectance-Fourier Transform Infrared spectroscopy (ATR-FTIR) was used to determine the changes in specific chemical bonds due to the plasma treatment in different working gases on maize grains. Measurements were carried out using spectrometer Bruker Vector 22 with Pike MIRacle accessories . The spectra were acquired in the range 4000–500 cmThe wettability of the grain surface was evaluated by measurement of water contact angle (WCA) by Drop Shape Analyzer DSA30  with software DSA3. The contact angles of distilled water droplets with a volume of 2 µL were measured, and the average value was obtained from at least 10 measurements.For one experimental run, dry maize grains (50 grains for each variant) were weighed on analytical scales and soaked in sterile distilled water for 4 h at room temperature. Imbibed grains were blotted dry, weighed again, and wrapped in wet sterile filter paper. Rolls were cultivated in dark conditions in an incubator at 24 ± 2 °C for 5 days. During cultivation, the number of germinated grains was counted, and material (seedlings without endosperm) for biochemical analyses was collected after 3 days. After 5 days, the length and weight of shoots and roots of young seedlings were measured. Obtained data were used to calculate germination (%), germination potential (%), germination index (%), grain vitality index (%), seedling vitality index (%), and seedling length index according to Abdul-Baki and Anderson .g), the supernatant was used to determine protein concentration according to Bradford [Samples (~1.5 g) were ground in liquid nitrogen with mortar and pestle and suspended in 50 mM Na-phosphate protein extraction buffer with 1 mM EDTA, pH 7.8. After 15 min centrifugation  as a substrate according to the methodology by Somogy  and Nelsá et al. . The abs2− (superoxide dismutase (SOD), E.C.1.15.1.1) and H2O2 (guaiacol peroxidase (G-POX), E.C.1.11.1.7) was monitored. The activity of SOD was established according to Beauchamp and Fridowich [−1 mg−1 of soluble proteins. Guaiacol peroxidase activity was expressed in µM of tetraguaiacol min−1 mg−1 of soluble proteins by molar extinction coefficient of tetraguaiacol 26.6 according to Chance and Maehly [The activity of enzymes that detoxify ˙Oridowich  and the ridowich . One unid Maehly . Chemica2HPO4, 4 mM NaH2PO4, 50 mM EDTA, pH 7) set on ice. For each sample, two roots were used, approximately one centimeter from the apex but excluding the root apex. The suspension of nuclei (100 µL) was mixed with 100 µL of 1% low melting point agarose (LMP), pipetted onto slides precoated with 1% normal melting point agarose, and covered with a coverslip. The nuclei fixed in LMP agarose on microscopic slides were lysed in cold high salt buffer  for 20 min. Subsequently, electrophoresis was performed in 1x TBE  at 1 V/cm at room temperature for 6 min. The slides were then dehydrated for 5 min in 75% ethanol and 5 min in 96% ethanol, left to dry and stored for subsequent evaluation. Nuclei were stained with 15 µL of 0.05 mM ethidium bromide for each mini gel on the slide. The stained nuclei were observed using fluorescence microscope OLYMPUS BX 51 with a green excitation filter UMWIG3 under 200× magnification and at least 100 nuclei per slide were evaluated by the visual scoring method [A neutral version of the comet assay was performed. The nuclei were isolated from 3-day-old maize seedlings using the slicing method with a sharp razor blade in 150 µL of ice-cold PBS -modified comet assay was based on a neutral comet assay described before with the Fpg additional steps according to Horváthová et al. . BrieflyAs a negative control (NC), seedlings not pre-treated with CAPP or any other agent were used.® Universal qPCR Master Mix  in thermocycler LightCyclerII 480  with the set of primers listed in All the experiments from RNA extraction to real-time PCR were performed by available commercial kits according to the manufacturer’s instructions. RNA was isolated from maize seedlings that were treated with CAPP generated in ambient air as grains, at 24, 48 and 72 h after sowing. For each sample, 2–3 seedlings without endosperm were used. Seedlings were immediately frozen in liquid nitrogen and ground with mortar and pestle. Total RNA was isolated using Spectrum Plant Total RNA Kit . The RNA samples were treated with DNase I enzyme, purified and concentrated using RNA Clean & Concentrator™—5 columns . The concentration and purity of total RNA were measured with spectrophotometer Nanodrop 1000 . The RNA integrity was checked by denaturing agarose gel electrophoresis . Using iAs reference genes, β-tubulin and elongation factor EF1α were used with the set of primers listed in http://www.ncbi.nlm.nih.gov/tools/primer-blast/; accessed on 4 November 2020) from sequences in the NCBI database.Primers were designed using the online tool Primer-BLAST .Fluorescent quantification of products in individual samples took place during polymerization using SYBR/FAM filter. The specificity of PCR products was confirmed by agarose gel electrophoresis and melt curve analysis, which was included at the end of each run of the qPCR reaction. The sequencing of PCR products was performed by the Sanger method at the Department of Molecular Biology . Sequences were analyzed using the program Sequencing Analysis software  and verified by the Blast bioinformatic tool  and Statgraphic Centurion 19 . Treatment effects were investigated by means of ANOVA single-step multiple comparisons using LSD test, and comparisons between the mean values were considered significant at 2 transformed normalized expression per sample.Relative change in gene expression was estimated according to Taylor et al.  includinCold atmospheric pressure plasma has a potential to be used in agriculture for seed treatment for more environmentally friendly production of various crops. Until then, it is necessary to study its effects on various plants to gain a holistic view and to be able to design optimal treatment conditions for the desired outcomes.The presented work focused on the effects of CAPP treatment of maize grains. We found that after the treatment, the wettability of grain surface increased and also the amount of water taken up by the grains during imbibition was higher. The surface diagnostics using ATR-FTIR did not show damage of the surface chemical bonds of the grains after plasma treatment. The CAPP generated by DCSBD plasma source is characterized by high volume power density, so short plasma treatment times are enough to achieve a substantial improvement of seed germination and the vitality of grains and young seedlings. However, higher doses of CAPP (90 or more seconds) had more of a negative impact on growth and production indexes. Specifically, higher doses of CAPP generated in nitrogen completely inhibited germination. The increased amount of soluble proteins was observed after the plasma treatment which can be caused by mobilization of storage proteins by proteases. The activities of antioxidant enzymes in variants treated with air and oxygen plasma were lower or comparable to the negative control, suggesting that the oxidative stress in three-day-old maize seedlings, which were treated with CAPP as grains, was not that high. Prolonged exposure to nitrogen plasma led to a reduction (N90) or complete inhibition (N120–300) of germination, which could be due to excessive oxidative stress and the inability of the antioxidant system to buffer it. A slight increase in DNA damage caused by CAPP treatment was detected, but mostly for longer treatment times. There was also a noticeable level of oxidative DNA damage, especially for ambient air-generated CAPP, however, these were primary DNA damage and cells are capable repairing them. We also found that HSF17 is probably not needed for the regulation of HSPs expression in response to CAPP treatment. On the other hand, the expression of HSP101 and HSP70 genes was induced by CAPP treatment, mostly at the first 48 h after sowing, which suggested that the quick action of molecular chaperones is needed shortly after CAPP treatment.


These topical feline endectoparasiticide products were tested for efficacy against fleas and intestinal helminths in a field trial in Brazil. Flea- and/or helminth-infested domestic cats were treated twice at a monthly interval following label instructions: 160 cats with Frontline® Protect/Broadline® and 165 cats with NexGard® Combo. The flea and intestinal helminth infestations were evaluated using comb counts and copromicroscopy, respectively before first treatment for baseline value, then 9 and 30 days after each treatment for fleas, and 9 days after each treatment for helminths. Multiparasitism was very frequent at baseline, as amongst the 325 included cats, 295, 280, 86 and 93 cats were at least infested with Ctenocephalides fleas, Ancylostoma, Toxocara and Dipylidium caninum, respectively. Efficacies were calculated by comparing the geometric means at baseline and at post-treatment timepoints for each parasite genus/species. Inclusive of both products and of all evaluation timepoints, the Ctenocephalides, Ancylostoma, Toxocara and D. caninum efficacies were at least 98.3%, 99.8%, 99.8% and 96.3%, respectively. No adverse reactions were observed, except for a few instances of mild, transient, and self-resolving hypersalivation occurring on the day of treatment in both groups. This field trial demonstrated high-level efficacy of Frontline® Protect/Broadline® and NexGard® Combo against major parasites of cats in Brazil.Eprinomectin and praziquantel, nematodicide and cestodicide compounds, are both combined with the insecticide and acaricide compounds fipronil and (S)-methoprene in Frontline The geometric mean was calculated by taking the anti-logarithm of the average of the log-counts and then subtracting 1, or was computed by taking the anti-logarithm of the least square mean minus 1 from the analysis model. Flea efficacy was also calculated on the basis of arithmetic means.Total live flea counts, EPG and proglottid counts were transformed to the natural logarithm of (count + 1) for analysis and calculation of the geometric means. The percent efficacy was calculated using the formula 100 × [, and the percent reduction of live fleas compared to baseline, 9 and 30 days after each of the two treatments ranged from 98.3% to 99.7%.In the Broadline® Combo group, the 149 cats evaluated for fleas were infested with 6–31 fleas at inclusion (geometric mean 10.9), and the percent reduction of live fleas compared to baseline, 9 and 30 days after each of the two treatments ranged from 99.0% to 99.9%.In the NexGardThe intestinal helminths efficacy results are presented in ® group, the helminth efficacies evaluated nine days after each treatment exceeded 99% for nematodes (Ancylostoma and Toxocara) and 97% for Dipylidium.In the Broadline® Combo group, the helminth efficacies evaluated nine days after each treatment exceeded 99% for nematodes (Ancylostoma and Toxocara) and 96% for Dipylidium.In the NexgardAt inclusion, inclusive of both groups, most cats were infested by fleas (91.0%), by helminths (94.8%), and were mixed infested by fleas and helminths (85.8%). The types and proportions of infestations including the mixed infestations are summarized in ® and 330 NexGard® Combo treatments were administered.A total of 320 Broadline® group, nine occurrences of mild and self-resolving hypersalivation were observed and lasted up to 40 min in eight instances, and 24 h one instance. No other adverse reactions were observed.In the Broadline® Combo group, five occurrences of mild and self-resolving hypersalivation were observed and lasted up to 40 min. No other adverse reactions were observed.In the NexGardNo animal needed or received any concurrent medication during the study.® or NexGard® Combo applied to cats in Brazil have a comparable and high level of efficacy against fleas, Ancylostoma and Toxocara infestations. The results also showed high efficacy against D. caninum infections for both products, however, less conclusively as the detection methods used  lack sensitivity and have been demonstrated to only reveal a fraction of the true infection incidence when compared to worm counts performed under necropsy [® Combo, is a novel systemic compound to which no fleas have yet been exposed in Brazil, while on the other fipronil, the ectoparasiticide active ingredient of Broadline®, is a contact compound that has been used worldwide for several decades, including in Brazil, in several animal species, public hygiene, and also in crop agriculture. The high efficacy results of fipronil (in combination to (S)-methoprene) observed in the present field trial show that this ingredient remains a good tool to control fleas in Brazil.This field trial confirmed that Broadlinenecropsy . ComparaAelurostrongylus abstrusus and Troglostrongylus brevior, major lungworms of cats were not investigated. The specific diagnostic technique for A. abstrusus and T. brevior (Baermann funnel migration) was not performed due to study logistics; nevertheless, the worldwide importance [This trial also confirmed that cats are a species commonly infested by multiple parasites, as referenced in Brazil , 19, 23 portance , 29, 30 portance , 22 of tThese observations confirm the importance of using a broad endectoparasiticide medical strategy in cats, including ectoparasiticide, nematodicide and cestodicide treatments.® and NexGard® Combo against major parasites of cats in Brazil.This field trial demonstrated a high level of tolerance and efficacy of BroadlineThe work reported herein was funded by Boehringer-Ingelheim. The authors are current employees or contractors of Boehringer-Ingelheim. Other than that, the authors declare no conflict of interest. This document is provided for scientific purposes only. Any reference to a brand or trademark herein is for information purposes only and is not intended for any commercial purposes or to dilute the rights of the respective owners of the brand(s) or trademark(s).


In this study, emulsion gel beads for loading quercetin were prepared through an emulsification/gelation process using whey protein isolate (WPI) and pectin. Emulsion gel beads’ properties were investigated by different pectin content. Additionally, the physicochemical properties, morphology and quercetin release properties from beads were explored. Firstly, electrical characteristics and the rheology of bead-forming solutions were measured, revealing that all systems had strong negative charge and exhibited shear-thinning behavior. The textural results demonstrated that the properties of emulsion gel beads were improved with increasing the content of pectin. It was also confirmed that crosslinking was formed between WPI emulsion and pectin by Fourier Transform Infrared (FTIR) analysis and thermogravimetric analysis (TGA). In addition, the shape of the beads was spherical or ellipses with smooth surfaces and they had a tight gel network of internal structures, which was visualized by using electron microscopy (SEM). Finally, the amount of quercetin released in vitro was gradually decreased with increasing pectin content; it was as low as 0.59%. These results revealed that WPI emulsion–pectin gel beads might be an effective delivery system for quercetin as a colon target and are worth exploring further. Quercetin , a plant flavonoid, exists in a variety of vegetables, fruit and grains. Recently, quercetin has attracted considerable attention due to its beneficial biological activities, such as antioxidative, anti-inflammatory, antibacterial and anticancer effects ,2. HowevRecently, several delivery systems have exhibited that the bioaccessibility of quercetin can be improved, including emulsions and hydrogels, etc. ,5,6. EmuHydrogels can absorb a great deal of water due to their three-dimensional network structure, making them ideal materials for functional food or pharmaceutical applications when they are swelled under physiological environments . Among tIn fact, proteins ) and polysaccharides  have been widely investigated as gelling agents. WPI contains more positively charged amino acids (267.9 mg/g) at pH 7.0 ), which are easy to react with anionic polysaccharides through electrostatic attractions. Moreover, WPI has higher solubility and surface hydrophobicity. Several reports have shown that protein solubility affects the interactions between protein and water, and surface hydrophobicity has an impact on protein–oil interactions on droplet surfaces . Simultaneously, pectin can improve the stability of emulsion through slowing down the movement of droplets, decreasing the collision frequency and coalescence rate, which are attributed to increasing the viscosity or forming a gel network in continuous phase. Therefore, the purpose of this study was to investigate the properties of emulsion gel beads loaded with quercetin. WPI and pectin were used as emulsifiers and structural strengthening agents, respectively. The encapsulation of quercetin in WPI emulsion–pectin gel beads was characterized and the release behavior of quercetin in the simulated gastrointestinal tract was estimated. The developed emulsion gel beads will have broad application prospects for delivering and controlling the release of active substances in the food and pharmaceutical industries.− when dissolved in water [The electrical characteristics of WPI emulsion–pectin complex were tested because these determine the nature of the electrostatic interactions between biopolymers . All beain water .In order to assess the rheological properties of the WPI emulsion–pectin complex solutions whether impacted the formation of beads or not, their apparent viscosity were measured b. There The texture characteristics of beads were measured. Hardness is regarded as the force required to attain a certain deformation, and it serves as a measure of food firmness . As show−1, which represented the carbonyl (C=O) stretching of the secondary amide (amide I band), the bending vibrations of the N-H (amide II band) and the N–H stretching (amide III band), respectively. The absorption bands of pectin was detected at 3394, 2926, 1746, 1411 and 1048 cm−1 corresponding to the stretching vibrations of hydroxyl groups, the stretching vibrations of –CH bonds of methyl esters groups or pyranoid ring carbons, the carbonyl (C=O) stretching of the methyl esterified carboxyl groups, asymmetric stretching vibrations of –C–O–C– bonds and −CH groups or methyl esters groups and skeletal C–O and C–C vibration bands of glycosidic bonds and a pyranoid ring, respectively [−1, which were stood for –OH stretching, aromatic ketonic carbonyl stretching, aromatic bending, C=C stretching of the aromatic ring, –OH phenolic bending, and aromatic stretching, respectively [As shown in ectively ,26. As fectively ,28.−1, suggesting that the cross-linking existed between WPI and pectin. In addition, the peaks at 1633 and 1411 cm−1 of pectin were shifted to around 1650 and 1460 cm−1, respectively, indicating that carboxyl groups (–COO−) were cross-linked with Ca2+ [−1 which was a characteristic peak of quercetin . However, it was interested that the peak of 80% pectin beads shifted to 1327 cm−1 because the fraction of WPI emulsion decreased.Furthermore, it was clearly shown that a new peak of 20%, 40%, 50%, 60% and 80% pectin beads was appeared around at 2850 cmith Ca2+ . The encA thermogravimetric analysis of all beads as shown in the The swelling degree is a crucial factor in the release of drugs from delivery systems . Gel beaThe visual appearances, surface and tangent plane morphologies of all beads were determined. The WPI emulsion–pectin gel beads with different concentrations of pectin  for the quercetin delivery system were prepared by an ionic gelation method. The particle sizes of freshly prepared beads depended on the pectin content, and their diameter was significantly increased when the pectin content increased from 20% to 80% . ResearcThe differences in the surface morphology and internal structure of quercetin containing beads (after they were lyophilized) from observing SEM micrographs exhibited in The content of quercetin in WPI emulsion–pectin beads was measured. Compared with WPI emulsion, the encapsulation efficiency of all beads was improved a. The enThe in-vitro release study was aimed to reveal that quercetin could be released slowly from the beads. It was found that the release of quercetin from all samples was below 10% after exposure to simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) in the In this study, a core-shell bioactive delivery system using natural food grade materials was successfully constructed. Beads were fabricated by mixing WPI emulsion with pectin under the ionotropic gelation method. FTIR, TGA and texture methods were used to investigate the effect of pectin content on the properties of beads the gel beads, confirming that incorporation of high pectin content into gel beads affect their texture, mesh-like structure and thermodynamics. These findings are important for producing emulsion gel beads during gelation because the properties of emulsion gel beads may affect encapsulation, stability, and release of hydrophobic functional ingredients embed in emulsion gel beads. As for in vitro digestion, it was found that beads loading quercetin were stable in SGF and SIF, indicating indirectly that it might be a promising delivery system to colon target for quercetin release. Nonetheless, the delivery systems for colon target should be further explored deeply. To sum up, these results might provide valuable information for nutraceutical delivery systems in functional food industry.2) were bought from Sinopharm Chemical Reagent Co., Ltd. . All chemicals were of analytical grade and the water used in this study was deionized.Whey protein isolate (WPI) was purchased from American Davisco chemical co., LTD. The product contained 97.6% protein (dry basis), as determined by the supplier’s standard proximate analysis procedures. Medium-chain triglyceride (MCT) oil and low-methoxy pectin were bought from Yuanye Bio-Technology Co., Ltd.  and Solarbio Co. Ltd. , respectively. Quercetin, pepsin, lipase, pancrelipase and bile salts were purchased from Sigma-Aldrich . Sodium chloride (NaCl), hydrochloric acid (HCl), sodium hydroxide (NaOH) and calcium chloride (CaClThe dispersions of WPI (10 wt%) and pectin (2 wt%) were stirred at room temperature for 2 h using a magnetic stirrer, respectively, and then pH values of dispersions were adjusted to 7.0 with 1 M HCl and NaOH. These solutions were kept overnight to ensure complete dispersion and dissolution.For production of quercetin emulsion: WPI stabilized quercetin emulsion (10 wt% protein) was prepared with 10 wt% medium chain triacylglycerol (MCT) oil containing quercetin  as the dispersed phase and 90 wt% aqueous phase solution at room temperature. The quercetin emulsion was prepared using an Ultra-Turrax  at a speed of 10,000 rpm for 3 min to form coarse emulsions, which were subsequently homogenized by using an ultrasonic cell disruptor  at ultrasonic power was 300 W for 10 min.w/w) calcium chloride solutions by using 5 mL measuring pipets and the distance between the tip of pipet and the surface of CaCl2 solutions was fixed at 10 cm, respectively. The samples were allowed to gel in CaCl2 solutions for 30 min with mild magnetic stirring, and then beads were rinsed with distilled water.For production of emulsion gel beads: the above emulsions mixed with pectin solution  at room temperature for 1 h, and then they were dropped into 2% . All stock solutions were diluted to 500 folds using distilled water (the resistivity was 15 MΩ cm) to avoid multiple scattering effects. After loading the samples into the instrument, they were equilibrated for about 120 s before particle charge data was collected over 11 continuous readings.−1.The apparent viscosity of stock solutions was measured by using a dynamic shear rheometer  with a parallel plate geometry  at 25 °C. The samples were placed on the plates and equilibrated for 2 min before testing. The apparent viscosity of sample was calculated when shear rates were ranging from 2 to 200 sThe texture profile analysis (TPA) tests were performed by using a texture analyzer  equipped with a cylindric shaped probe . The freshly prepared spherical beads (about 2 g) were placed on a fixed bottom plate under the probe at 50% compression. The pre-test, test and post-test speeds were set at 1.0 mm/s, 0.5 mm/s and 1 mm/s, respectively. Furthermore, trigger force was 1 g, and the time between two loadings was 5 s. The texture parameters including hardness, springiness, cohesiveness, resilience, gumminess and chewiness values of beads are calculated by the software Texture Expert Version 1.22.The water content of beads with different pectin content was detected with Moisture Analyzer . The fresh gel beads (about 0.8 g) were placed on the plate made by aluminized paper for measuring.−1 and 60 scans at a resolution of 4 cm−1, and all spectra of the prepared disks were recorded.Fourier transform infrared spectroscopy (FT-IR) spectra was used for detecting WPI, quercetin, pectin and freeze-dried beads. All freeze-dried samples were examined on an FT-IR spectrometer . Firstly, all samples should be mixed with KBr reagent (a mass ratio of 1:50) and were ground into powders. Next, the powders were compressed into disks by using Hydraulic press  at 1.7 tons, which are used for FTIR detection. The wavenumber of FT-IR spectrum was between 4000 and 400 cmThe thermal stability of freeze-dried beads was measured by using Thermogravimetry Analyzer . The sample was placed on a platinum pan and the temperature was raised from 30 °C to 500 °C and the heating temperature was scanned at the rate of 10 °C/min. The weight loss was monitored with respect to time and temperature.s and Wi are the weight of the swollen beads and initial beads, respectively.The swelling behavior of the freshly prepared beads was detected according to the methods previously described ,37. BrieThe morphologies of freeze-dried gel beads were analyzed by a scanning electron microscope  using an accelerating voltage of 5 kV under low vacuum conditions. The surface and internal (cut with a blade) morphologies of freeze-dried gel beads were examined. The beads were affixed to conductive carbon tapes and coated with a thin layer of gold by a sputter coater before observation. Moreover, gel beads diameters were measured using Leica Application Suite EZ software 3.4.0.d is the quercetin concentration that remained in the sodium citrate solution and Ci is the initial quercetin concentration.The amount of quercetin present in beads was quantified using an Ultraviolet spectrophotometer . Before measuring, all beads were added into 2% sodium citrate solution until they were dissolved completely. Subsequently, referring previous study to measure the amount of quercetin . BrieflyThe quercetin-loaded gel beads were used to explore the release behavior in vitro digestion. The method was according to previous research . Briefly


The reduction of inequality is a key United Nations 2030 Sustainable Development Goal . Despite marked disparities in radiological services globally, particularly between metropolitan and rural populations in low- and middle-income countries, there has been little work on imaging resources and utilization patterns in any setting , Insights Imaging 6:573-7, 2015; Maboreke et al., An audit of licensed Zimbabwean radiology equipment resources as a measure of healthcare access and equity, 2020; Kabongo et al., Pan Afr Med J 22, 2015; Skedgel et al., Med Decis Making 35:94-105, 2015; Mollura et al., J Am Coll Radiol 913-9, 2014; Culp et al., J Am Coll Radiol 12:475-80, 2015; Mbewe et al., An audit of licenced Zambian diagnostic imaging equipment and personnel, 2020). To achieve equity, a better understanding of the integral components of the so called “imaging enterprise” is important. The aim was to analyse a provincial radiological service in a middle-income country.An institutional review board-approved retrospective audit of radiological data for the public healthcare sector of the Western Cape Province of South Africa for 2017, utilizing provincial databases.We conducted population-based analyses of imaging equipment, personnel, and service utilization data for the whole province, the metropolitan and the rural areas.2). Rural imaging facilities by population are double the metropolitan (20 vs 11/106 people). Metropolitan imaging personnel by population (112 vs 53/106 people) and equipment unit (1.7 vs 0.7/unit) are more than double the rural. Overall population-based utilization of imaging services was 30% higher in the metropole (289 vs 214 studies/103 people), with mammography (24 vs 5 studies/103 woman > 40 years) and CT (21 vs 6/103 people) recording the highest, and plain radiography (203 vs 171/103 people) the lowest differences.Metropolitan population density exceeds rural by a factor of ninety (1682 vs 19 people/kmDespite attempts to achieve imaging equity through the provision of increased facilities/million people in the rural areas, differential utilization patterns persist.The achievement of equity must be seen as a process involving incremental improvements and iterative analyses that define progress towards the goal. The reduction of inequality is a key United Nations 2030 Sustainable Development Goal (SDGs) , 2. DiagThe expense of modern diagnostic imaging has the potential to compound existing worldwide inequalities in access to radiological services. At one end of the spectrum are high-income countries with an aging population, a high burden of non-communicable diseases and a relative abundance of the more capital intensive imaging equipment such as mammography, computerised tomography (CT), magnetic resonance imaging (MRI) and digital subtraction angiography (DSA) , 18. At There has been limited in-depth work on radiological resources and service utilization patterns globally, particularly in LMICs. Although the WHO has published national estimates of high-end medical imaging resources based on questionnaire surveys of member countries, these data do not include the more basic and affordable equipment . FurtherSA is one of five upper middle-income countries in sub-Saharan Africa. It has adopted the District Health System (DHS) for delivery of comprehensive primary care in the public healthcare sector . Health metropolitan or rural streams , healthcare facility and provincial service stream . All equipment units included in the study were fully functional and serviced regularly. The single metropolitan stream consists of the City of Cape Town, while the parallel rural stream includes five districts namely West Coast, Cape Winelands, Overberg, Garden Route and Central Karoo. Rural referral pathways from District to Regional Hospital breach individual district boundaries, precluding analysis by individual rural district . Plain rThe number of radiology equipment units per million people was calculated by modality. The quantum of radiological examinations for each modality was assessed per thousand people, as well as per thousand patient engagements. Imaging examinations per equipment unit were calculated by modality.The numbers of registered personnel in the categories of diagnostic radiographer, sonographer, registrar, and specialist radiologist were collated and categorized by healthcare facility and service stream.The study was approved by the Head of Health of the Western Cape Government (WCG), the WCP Health Research Committee, under the auspices of the National Health Research Database and the Health Research Ethics Committee of Stellenbosch University.2) exceeds rural (19 people/km2) by a factor of almost ninety (89:1)  provincial imaging facilities. All have plain radiography and fifty-three  have ultrasound services. The metropolitan and rural areas have 35 and 33 imaging facilities, respectively. As a result, rural resources by population are almost double the metropolitan (20 vs 11/106 people)  and Community Day Centres  together account for almost three-quarters  of all provincial facilities, with Community Day Centres predominant  in the metropole and District Hospitals  most common in the rural areas. All Central Hospitals (n = 3) and Community Health Centres (n = 10) are in the metropole, while the six Regional Hospitals are equally distributed between the metropolitan and rural areas , ultrasound machines approximately a quarter  and CT scanners less than 5 %   and ultrasound (24.7 vs 14.5/106 people) resources exceed metropolitan by 17 and 70%, respectively. Additionally, rural access to mammography is almost triple that in the metropole (14 vs 5 units/106 women > 40 years)  are in the metropole and more than three-quarters  are radiographers  and equipment unit (1.7 vs 0.7/106 people) are more than double the rural. Of note, the rural areas have more equipment units than personnel  by radiologist at provincial, metropolitan and rural levels was 35,820, 30,219 and 63,825 studies, respectively (Table 3 people and 1.3 investigations/patient. The CXR (n = 92/103 people) was the commonest single examination, while plain radiography accounted for almost three-quarters  of all investigations. Radiography  and ultrasound  together constituted almost 90 %  of all provincial investigations , with mammography  and CT  recording the highest differences and plain radiography utilization  the lowest. However, overall patient-based utilization of services in the rural areas exceeded that in the metropole by 20% (1301 vs 1080 studies/103 patients) reflecting increased plain radiography and ultrasound usage  and fluoroscopy (n = 450 vs 140/unit). Seventy percent of all equipment usage was confined to normal working hours. This figure was similar for the province, the metropolitan and the rural areas . The highest differences were in mammography  and equipment unit (1.7 vs 0.7 staff/unit) are more than double the rural. Furthermore, the number of rural equipment units exceeds personnel.Firstly, the number of rural imaging facilities and equipment units per million people exceeds that in the metropole. This represents a commitment on the part of the WCP DoH to enhance access to imaging for the relatively sparse rural population. However, metropolitan personnel resources by population (3 people) was 30% less than metropolitan (289 studies/103 people). Despite the rural areas being 17, 70 and 263% better resourced for plain radiography, ultrasound and mammography, respectively, utilization of these modalities was just 74, 45 and 7% of that in the metropole. Imaging utilization thus appears to represent a complex interplay between various determinants within the imaging enterprise. This is further illustrated by the finding that plain radiographic and ultrasound examinations per patient were 23 and 10% higher, respectively, in the rural areas compared to the metropole. The intuitive explanation is that this reflects lower CT availability in the rural districts (1.7 scanners/106 people) compared to the metropole (3.2 scanners/106 people). The inference is that if there is not ready access to CT, a more expensive modality with higher diagnostic sensitivity and specificity, recourse is made to the more basic, affordable, and accessible option. Conversely, there may also be a concurrent lower threshold for metropolitan CT utilization, given the more ready availability of the modality.Secondly, notwithstanding the higher rural imaging facility and equipment resources by population, overall rural service utilization  and plain radiography (n = 5377) achieved the highest average annual outputs per unit across the province, while fluoroscopy recorded the lowest (n = 347), being approximately 17-fold below that of CT. The limited use of fluoroscopy reflects declining global trends and suggests that provincial policy on fluoroscopic service provision merits review. Of note, less than 3 fluoroscopic investigations/103 people were performed across the province in the review period.This work also provides novel insights into the differential equipment workload across the modalities and regions. CT (n = 4365) achieved 72% of metropolitan output (n = 6058), whilst the greatest difference was in mammography, with rural output (n = 458) a mere 7% of metropolitan (n = 6198). The optimal combination of equipment, personnel, and hours of operation for rural facilities remains a conundrum. WCP rural equipment resources currently exceed imaging personnel by 46% (n = 139 vs 95), likely contributing to decreased equipment utilization. A recent study of Zambian radiological equipment and personnel resources [Rural equipment utilization by unit was less than metropolitan across all modalities. The smallest differential was in plain radiography, where the average rural unit  this would be more than 5-fold the average caseload  documented for general radiology consultants working in the United States [n = 8171) in the United Kingdom (UK) [This is the first detailed appraisal of the potentially overwhelming workload of the rural radiologist. If all imaging studies performed in the rural areas were to be formally reported by a radiologist (d States  and moredom (UK) . It is cThe challenge of meeting the ever-increasing demand for radiology reporting is not unique to lower resourced environments , 39. A r2) and Finnmark (1.6 people/km2) were the counties with the highest and lowest population densities, respectively [3 people) was double that in the more sparsely populated Finnmark (459 studies/103 people). By comparison, there was a 1:1.8 differential in plain radiographic utilization between metropolitan (203 studies/103 people) and rural (171 studies/103 people) areas of the WCP.To the best of our knowledge, the only comparable analysis of geographic variation in the utilization of radiographic services was conducted in Norway in 2002 . The stuectively . Plain rn = 581) and Romania (n = 143) have the highest and lowest plain radiographic examinations per 103 people, respectively, while the WCP (n = 192) utilization is at the lower end of this range. WCP plain radiograph utilization is most closely aligned with Sweden (n = 201/103 people) and Denmark (n = 180/103 people).The increasing population dose from medical exposures to ionising radiation is a global concern . This stn = 271) and Costa Rica the lowest (n = 34) CT examinations per 103 people. CT utilization in the WCP (16/103 people) is less than half that of Costa Rica, also an upper middle-income country [Our analysis also allowed comparison of WCP CT utilization with global data , 41. The country .Our study provides a compelling argument for upscaling undergraduate medical education programs in the interpretation of plain radiographs and the performance of basic ultrasound examinations. Equipped with such skills, medical graduates would be well-placed to address approximately 90 % of provincial reporting needs. Additionally, competence in basic interpretation of plain radiographs would address the current reporting void in this modality. Our study also shows that educational initiatives in CXR interpretation would be particularly beneficial, since almost half of all plain radiographic studies are CXR.This study’s major strengths are it being the first analysis of radiological utilization patterns in a middle-income country, its inclusion of all components of the so-called “imaging enterprise”, and the differentiation of rural from metropolitan services. In addition, the study focused exclusively on the utilization of public-sector resources, which provide services to the uninsured population, the more vulnerable members in our society. Published studies to date have been confined to high-income countries, have not correlated utilization with resources, and have not always drawn a clear distinction between private and public-sector resources and utilization patterns.The study was limited by its retrospective design. However, this was mitigated by the meticulous collection and collation of all healthcare facility data in standardised format. An additional limitation was the inability to accurately assess the ultrasound workload of the various categories of personnel within the radiology domain, since outputs were not stratified by healthcare worker. In the WCP, ultrasounds in the radiological domain may be performed by either specialist radiologists, radiology registrars, sonographers, or dual-qualified radiographers, the latter performing both radiography and ultrasound. While radiologists, dual-qualified radiographers and sonographers practice independently, registrars generally report under specialist supervision. A further limitation is that many provincial obstetric and gynaecological ultrasounds, as well as point of care ultrasounds, are not performed within the radiological domain and are not included in this analysis. Going forward, WCP ultrasound data would be enhanced by allocating outputs by category of healthcare worker and including data from outside the radiological domain.The study highlights WCP attempts to bridge the divide between rural and metropolitan access to imaging, through the provision of an increased number of imaging facilities and basic equipment units per million people in the rural compared to metropolitan areas. However, it also underscores the complexity of achieving equitable utilization of services between rural and metropolitan areas.The achievement of equity in all aspects of healthcare must be seen as a process, involving incremental improvements and iterative analyses that define progress towards the goal. Studies such as this serve to define the baseline and inform future interventions to enhance equity going forward.


Thrombolysis for acute ischemic stroke (AIS) with alteplase is the currently approved therapy for patients who present within 4.5 h of symptom onset and meet criteria. Recently, there has been interest in the thrombolytic tenecteplase, a modified version of alteplase, due to its lower cost, ease of administration, and studies reporting better outcomes when compared to alteplase.This systematic review compares the efficacy of tenecteplase vs. alteplase with regard to three outcomes: (1) rate of symptomatic hemorrhage, (2) functional outcome at 90 days, and (3) reperfusion grade after thrombectomy to compare the efficacy of both thrombolytics in AISThe search was conducted in August 2021 in PubMed, filtered for randomized controlled trials, and studies in English. The main search term was “tenecteplase for acute stroke.”A total of 6 randomized clinical trials including 1675 patients with AIS was included. No one’s study compared alteplase to tenecteplase with all three outcomes after acute ischemic stroke; however, by using a combination of the results, this systematic review summarizes whether tenecteplase outperforms alteplase.The available evidence suggests that tenecteplase appears to be a better thrombolytic agent for acute ischemic stroke when compared to alteplase. Alteplase is the only the Food and Drug Administration (FDA) approved thrombolytic for thrombolysis for acute ischemic stroke (AIS). When given to eligible patients within 4.5 h, there is a 28% decrease in disability at 90 days, and a more rapid improvement is associated with greater symptom improvement . The risBoth alteplase and tenecteplase are thrombolytic agents that achieve their effect by binding to fibrin in clots and converting entrapped plasminogen to plasmin. Plasmin in turn breaks up the thrombus. Tenecteplase is a modified form of alteplase with three point mutations that renders it a larger molecule with a longer half-life . These pTenecteplase has been studied for over 20 years in the myocardial infarction (MI) population. When compared to alteplase for MI, tenecteplase showed equal vessel patency at 90 min  and equaThis systematic review focuses on the effects of tenecteplase compared to the effects of alteplase in treating patients with acute ischemic stroke. The objective of this review is to summarize the impact of both thrombolytics on (1) the rate of symptomatic hemorrhage, (2) the functional outcome of the patient after 90 days, and (3) the reperfusion grade after the thrombectomy.Strategies used to select studies, extract data, and make objective assessments based on both qualitative and quantitative information from the available literature were performed according to the Preferred Reporting Items for Systematic Review (PRISMA) guideline . A formaStudies were independently searched PubMed using the following search terms: “tenecteplase for acute stroke.” The search was limited to human randomized controlled trials. Only studies written in English were searched. Bibliographies of retrieved articles were manually checked for additional references. Studies were included if they met any of the following criteria: administration of tenecteplase for AIS and any discussion of (1) occurrence of symptomatic hemorrhage, (2) functional outcome at 90 days, and (3) reperfusion grade after thrombectomy. Functional outcome was assessed using the modified Rankin score (mRS). Reperfusion grade was assessed using the thrombolysis in cerebral infarction (TICI) perfusion scale, graded from 0 (no perfusion) to 3 (complete perfusion).The search was conducted in August 2021.Population: The study population included adults (18 years old or older) diagnosed with acute ischemic stroke confirmed by diagnostic guidelines updated by the American Heart Association/American Stroke Association .Intervention: Interventions had to be treatment-based, including tenecteplase or alteplase intravenously administered.Controls: Because the goal of systematic review is to compare the current medication utilized by physicians to a medicine not yet approved, there is no formal control. If anything, alteplase is the control of the studies because the goal was to assess if tenecteplase had a comparatively better, equal, or worse outcome.Outcome: Outcome measures had to assess the occurrence of hemorrhage, functional outcome (modified Rankin Score measured at 90 days), and reperfusion grade after thrombectomy.Study design: All designs had to be randomized controlled trials.Time: Articles considered included those published between February 2010 and August 2021.Studies were included if they met the following Population, Intervention, Controls, Outcome, Study (PICOS) criteria :PopulatStudies were excluded from the review if they were not written in English and if the authors had a bias for or against a given drug. In addition, pulmonary embolism and myocardial infarction studies assessing the combined effects of both drugs were also excluded from the review.We only included randomized control trials comparing the effects of the administration of tenecteplase 10–40 mg) to alteplase (90 mg) in patients with acute ischemic stroke. The primary efficacy endpoint analyzed was treated patients’ absolute risk of symptomatic intracranial hemorrhage (sICH) and functional ability at 3 months post-stroke, and their reperfusion grade if they underwent thrombectomy. Symptomatic hemorrhage events in trials were identified using the specific sICH definition clarified by that trial. Any discrepancies in the study selection were resolved by consensus. Full texts were retrieved and evaluated based on the previously set inclusion criteria. See Fig. 0–40 mg tData were extracted and documented based on predetermined criteria to identify solely relevant information. Details recorded from each reference include the author’s last name, study publication year, participants, intervention, outcomes measures, and results of the study. Data were extracted by one reviewer and checked for accuracy by the second reviewer. Each of the studies and patients’ characteristics and details of intervention is summarized in Table The risk of bias assessment was created based on the handbook of Cochrane (5.1 version) . To assen = 18), the type of study did not meet our inclusion criteria (n = 6), the article came up twice through the search (n = 3), or the study was prematurely terminated (n = 1). Six articles were included in the systematic review. See Fig. A total of 34 articles were retrieved in this search. Thirty-three articles were considered for this screening, and 28 articles were excluded because the study did not focus on stroke patients n = 8, the tyIn total, the data consisted of 1675 patients treated with intravenous thrombolytics for acute ischemic stroke. A total of 782 patients received tenecteplase while 727 received alteplase (not including 5 because of a lack of specification). Based on the available information, the mean age of the participants across 3 studies was 70.4 years SD = 14.4) for patients using tenecteplase and 71.3 SD = 15.9) for patients using alteplase, ranging from 49 to 92 years across all treatments –14. All .4 for pa.9 for paEach of the studies’ randomization, allocation blinding, incomplete outcome data, double-blinding, and other sources of bias were assessed as a low risk of bias in most of the included studies , 13, 16.Study objectives did not vary for most of the studies: tenecteplase versus alteplase for acute ischemic stroke –16, teneAll six trials involved a randomized, prospective, open-label, and blinded endpoint trial with prior baseline CT scans to first establish lack of intracerebral hemorrhage –17. In fPatients were not informed of treatment allocation. Within 24–48 h of treatment, symptomatic intracranial hemorrhage and intracranial hemorrhage were ascertained –16. OtheThe included studies Table  show thaP=0.82) [P=0.091) [P=0.002) [Four trials showed insignificant differences in the percentage of patients with symptomatic hemorrhage –14, 16. P=0.82) . CampbelP=0.091) . It may p=.98) [Two studies reported on functional outcome, and there was no statistically significant difference between tenecteplase and alteplase. Logallo et al. identified an excellent functional outcome for 64% of the patients in the tenecteplase pool and 64% of the patients in the alteplase pool (p=.98) . Roning p=.98) .P=0.002) [P=0.004) [In three trials, the criteria of “reperfusion rates after thrombectomy” was not mentioned, quantified, or the focus of their papers , 14, 16.P=0.002) . ParsonsP=0.004) . RonningAcute ischemic stroke remains the leading cause of disability worldwide and confers a significant burden to the quality of life for the stroke survivor and their caregivers. Because of this, there is tremendous interest in both targeted and supportive therapeutics to help ease this burden. Research has been done on the impact of acute blood pressure –20, antiTenecteplase also has additional benefits. It can be given as a single bolus, which is more comfortable for the patient, less arduous for the hospital staff, and certainly more convenient for prehospital or interhospital transfer. Tenecteplase is also currently cheaper. A 50 mg vial of tenecteplase costs approximately $6300 while a 100 mg vial of alteplase costs approximately $9200 .The overarching purpose of this systematic review was to evaluate the effectiveness of tenecteplase versus alteplase on AIS patients eligible for thrombolytic therapy. The results demonstrate that tenecteplase is just as good and in some cases better than alteplase with regard to the outcomes of (1) post thrombolytic bleeding, (2) functional outcome at 90 days as measured by the mRs, and (3) recanalization/reperfusion rates following thrombectomy.


P = 0.03). Thus, there is a remarkably high prevalence of an unhealthy pattern of beverage intake, and specific maternal factors may be implicated as enablers of such behaviors, which is also addressable for future interventions. Childhood and adolescence represent critical periods where beverage and food consumption behaviors are learned and developed. Mexican mothers’ presence and influence are instrumental in shaping such behaviors. The aim of this study was to estimate the prevalence and risk associations of maternal factors for unhealthy patterns of beverage intake. This study analyzed data from a population-based cross-sectional study of healthy children and adolescents from Mexico City. Data of subject’s total water intake (TWI) and its’ sources were collected using two 24-h recall surveys. Patterns of beverage intake were constructed based on the guidance system of beverage consumption in the US. Maternal factors of interest included age, body mass index (BMI), mother’s educational level (MEL), socioeconomic status (SES), and belongingness to the paid workforce (BPW). Data of 1532 subject–mother dyads informed that 47% of subjects did not meet the Institute of Medicine (IOM) recommendations for TWI, and 94.6% showed an unhealthy beverage intake pattern, mainly consisting in a lower intake of water and a higher intake of caloric beverages with some nutrients; and calorically sweetened beverages. The major sources of hydration were caloric beverages with some nutrients . The highest risk association for an unhealthy beverage intake pattern was seen in those subjects with mothers in the cluster with lower SES, lower MEL, lower proportion of BPW, higher BMI, and younger age (OR = 9.3, 95% CI 1.2–72.8,  Hydration status is a significant health indicator and directly correlated with healthy metabolism ,2. WaterWater intake behaviors vary among individuals depending on age, gender, culture, education level, and availability of water sources, among other factors . EvidencThe United States Institute of Medicine (IOM) has published recommendations for an adequate daily water intake for different sex and age groups . KnowledThe sources of water are fluids and water from food. The fluids are composed of different types of beverages and have been classified according to their nutritional and calorie contents by the “Guidance System for Beverage Consumption in the U.S.” . This clChildhood and adolescence represent critical periods where eating behaviors are learned and developed . These bThe aim of this study is to estimate the prevalence and risk associations of maternal factors for unhealthy pattern of beverage intake in Mexican children and adolescents.Data for the current analysis came from the “Reference Values of Body Composition in Pediatric Mexican Population” study, a population-based cross-sectional study of healthy children and adolescents from Mexico City ,17. Brie® 284  scale with stadimeter, measurements were standardized and performed by trained nutritionists. The body mass index (BMI) was calculated as the weight (kg) divided by the square of height (m) [2), underweight (BMI < 20 kg/m2), overweight (BMI 25 to <30 kg/m2), and obesity (BMI ≥ 30 kg/m2).Clinical measurements of subjects and their mothers included weight and height, measured with subjects wearing lightweight clothing using a SECAight (m) . Subjectight (m) . Mothers® . and for local products we used the published Mexican equivalents [Data on quantity and types of food and beverages intake was collected by trained nutritionists through a structured interview to each mother–subject dyad using two 24-h recall surveys, one administered to the nearest past typical school–day and the other to the nearest past typical weekend day. A weighted mean from both surveys was calculated for corresponding analyses. A 12-month food frequency questionnaire (FFQ) was also applied. Briefly the FFQ includes 133 food items in the following categories: dairy products, carbohydrates, fats, proteins, vegetables, fruits, water, beverages with and without added sugar, and highly processed calorie dense foods  . This FFivalents . Misreporting bias for this sample was performed applying the Goldberg cut–off method adapted for children by Black  and prevAll reported beverages and their quantities where captured and classified according to the following 6 levels :Level 1. Plain water: tap and bottled water.Level 2. Infusions: tea or coffee without sugar.Level 3. Low-fat & skim milk: skim milk, semi–skim milk, light milk, and low-fat drinkable yogurt without sugar.Level 4. Non-calorically sweetened beverages: infusions, flavored water, soft drinks, and industrialized drinks sweetened with non-caloric sweetener.Level 5. Caloric beverages with some nutrients: whole milk, flavored milk, sugary drinkable yogurt, natural juices, industrialized juices, sports drinks, industrialized prepared water, fruit water, and nectars.Level 6. Calorically sweetened beverages: soda, frappe drinks, energy drinks, infusions with sugar, and jelly.The beverage patterns were constructed based on the guidance system of beverage consumption in the US . This syAcknowledging a differential potential impact of complying or not to the different levels recommendations and incorporating the IOM and AHA recommendations on TWI and TASI respectively, we built a “suggested beverage intake score” (SBIS) and an “acceptable beverage intake score” (ABIS) with different weights for each of the recommendations. The IOM, AHA, level 1, and level 6 recommendations were prioritized and weighted as 20% each, followed by recommendation on level 5 beverages with 10%, and recommendations on level 2–4 beverages with 3.3% each. This approach could result in specific scores for each subject between 0 and 1, where 0 means not complying to any of the 8 recommendations and 1 complying to all of them. Differences between SBIS and ABIS consisted in the different criteria used for suggested or acceptable beverage intake patterns according to the guidance system of beverage consumption in the US . Data on mothers’ educational level (MEL) was collected and categorized as (a) elementary school (+6 years), (b) secondary school (+3 years), (c) high school (+3 years), (d) a technical degree (+1–3 years), (e) a bachelor’s degree (+4–5 years), and (f) a master’s or doctoral degree (+1–4 years). Socioeconomic status (SES) was determined as the total monthly income per family and categorized according to the classification of the “Asociación Mexicana de Agencias de Inteligencia de Mercado y Opinión (AMAI)” in Mexican pesos . This vaConventional descriptive statistics were used to inform regarding the characteristics of the subjects, their mothers and studied variables of TWI, frequency of beverages intake, TDEI, daily caloric intake coming from beverages, TASI and maternal factors. Because TDEI and TWI requirements differ by age and sex, data were disaggregated into three age groups: (a) young children (4–8 years old), (b) children (9–13 years old), (c) and adolescents (14–18 years), and each into males and females. The reported data on beverage consumption were used to estimate the total water intake (TWI), which was compared with the IOM age–specific recommendations for adequate intake , only foRelationships between maternal factors and non–compliance to the different recommendations, and SBIS/ABIS were assessed by univariate and multivariate logistic and linear regression analyses, adjusting for age and sex of the subjects.Two-step cluster analyses, taking the maternal factors MEL, SES, and BPW as independent categorical multinomial variables, mothers’ age and BMI as standardized continuous variable, and the number of clusters automatically determined based on log-likelihood distances were done. The resulting clusters were used to explore the specific combinations of maternal factors that may discriminate different risk associations to an unhealthy beverage intake pattern, lower SBIS and/or ABIS.® SPSS Statistics, version 20.0 Armonk, NY: IBM Corp. without replacement of missing values. Values of P  < 0.05 were considered statistically significant.All statistical analyses were performed using IBMThis study included data on 1532 mother–subject dyads. Sociodemographic and anthropometric data are summarized and presented in Eighty one percent of the sample consumed beverages from level 1, 3% from level 2, 17% from level 3, 7% from level 4, 94% from level 5, and 31% from level 6. A detailed summary of all types of consumed beverages disaggregated by age– and sex–groups is presented in Considering the total sample, 812 (53%) subjects comply with the IOM recommendations for adequate TWI. More than the half of the sample  exceeded the AHA TASI recommendation. Most of the subjects, a total of 1449 (94.6%) showed an unhealthy pattern according to non–compliance to neither of the suggested nor acceptable recommendations of the US Guidance System. Remarkably, only nine subjects (0.6%) showed a healthy beverage pattern, consisted with the compliance with the 8 recommendations . Details of these results disaggregated by sex and age groups are presented in Considering the total sample, major sources of hydration were levels 5  and 1 (water) with 46% and 34% of TWI, respectively. Level 6, 2, 4, and 3 beverages represented 7%, 4%, 2%, and 0.5% of TWI, respectively. Data for the different age and sex groups did not differ from the total sample. A graphic representation of this distribution for the total sample and disaggregated by age and sex groups is presented and contrasted to suggested and acceptable patterns in Median SBIS for the total sample was 0.5 (IQR 0.3–0.7), and for ABIS was 0.6 (IQR 0.47–0.7). Specific data on each age– and sex–groups are shown in Prevalence and distribution of maternal factors are summarized in P < 0.0001). In contrast, increasing MEL and BPW were associated to a decreased risk association for non–compliance to level–6 recommendation . BWP also showed a decreased risk association for non–compliance to level–1 recommendation . Finally, increasing maternal BMI showed a decreased risk association with non–compliance to level–5 recommendation . Multiple regression analysis of all maternal factors and adjusted by sex and age of the subjects for each of the 8 studied recommendations, confirmed a significant increased risk association of age of mothers for non–compliance to the IOM TWI recommendation . In contrast, age of mothers showed significant decreased risk associations for non–compliance for level 1 and 6 recommendations .Univariate logistic regression of maternal factors  for non-compliance to each of the eight recommendations indicated that increasing age of mothers showed a significant risk association for non–compliance to the IOM TWI recommendation . BWP kept statistically significant association for ABIS  and MEL showed only a tendency . Corresponding multivariate linear regression analyses adjusted for age and sex of the subjects showed no independent associations for any of the maternal factors.Univariate linear regression analyses of maternal factors to SBIS showed that MEL and BWP were significantly associated to higher scores . This observation kept statistical significance after adjusting for age and sex of the subjects . Details on each cluster are presented in A significantly higher risk association for an unhealthy beverage pattern was observed in those subjects with mothers belonging to cluster 1  of studied subjects did not comply to either suggested or acceptable recommendations from the Guidance System for Beverage Consumption in the U.S. The three more common recommendations that where not complied where not drinking enough level 1 beverages, and exceeding level 5 and 6 beverages. Seven hundred and twenty subjects (47%) consumed less than the IOM’s daily TWI recommendation and 829 (54%) exceeded the AHA daily TASI recommendation. Our results suggest that the beverage intake patterns in Mexican children and adolescents are in most cases inappropriate and may expose them to future risks of negative health outcomes. Other authors have previously reported mean daily TWI for Mexican children and adolescents with variable results. Barquera et al. analyzed nation–wide data from the Mexican National Health and Nutrition Surveys of 1999 and 2006 (ENSANUT) and reported a mean daily TWI of 794 mL for young children and 1254 mL for children, which are lower compared to our observations of 1457 and 1718 mL respectively . PiernasMore recently a multinational study that included data from Mexico, collected information on beverage intake through a validated liquid intake seven-day record . This stIrrespective of which of the previous discussed studies may reflect more precise data, all of them report consistently that Mexican children and adolescents report an average daily TWI below the age and sex–specific IOM recommendations . These observations also applied when we compared our data to those previously reported for adolescents by a multinational European study, where daily TWI was of 1611 and 1316 mL for adolescent males and females respectively, compared to our data of 1970 and 1750 mL, respectively .Interestingly, we observed a decreasing rate of compliance with the IOM recommendations for daily TWI in the older age groups, with 82% compliance in young children, and 60% compliance in adolescents. Similar findings were previously reported in an analysis of Mexican children and adult subgroups, wherein 60% of children and 43.3% of Mexican adults were reported as compliant . These fRegarding the sources of hydration, our main observations are consistent with other data informing that caloric beverages with some nutrients (level 5) are the major source of hydration of Mexican children and adolescents ,35. ThesAlthough in the level five include beverages with nutrients, as whole milk and 100% juices, the recommendation is not exceeding the consumption of this beverages because they have a high calories content that could lead to a higher daily caloric intake. According to the American Academy of Pediatrics, the recommendation for children and adolescents is to use non-fat (skim) or low-fat milk, and fruit juice offers no nutritional benefits over whole fruit for infants and children, and has no essential role in healthy, balanced diets.Univariate and multivariate lineal and logistic regression analyses that looked for potential independent associations between maternal factors and specific behaviors on beverages intake provided marginal associations with a few of them reaching statistical significance. Relevant to such findings McLeod et al. reported that Australian first–time mothers from major socioeconomic position had more knowledge about nutrition in comparison to those mothers with minor socioeconomic level . CampbelIn contrast to our findings, several authors, mainly from the United States of America, have reported that maternal fulltime employment may be associated with more unhealthy eating behaviors ,41. DiffP = 0.034). We believe that the defining characteristics of the two clusters of mothers identified in our sample, remark the relevance of societal disparities and their relationship to significant risk association to unhealthy beverages intake patterns. In our sample the combination seen for cluster 2 may speak of mothers that are less aware of healthier nutritional habits because of their lower education level, being at greater economic difficulties to provide appropriate care because of their lower SES, and less access to regular healthcare provided by social security because of their lower rate of BPW.Given the significant relationships between maternal factors, we considered of greater value the identification of specific combinations of these that may discriminate different risk associations to an unhealthy beverage intake pattern. The cluster analysis identified such groups with higher risk towards an unhealthy beverage intake pattern characterized by subjects with mothers of younger age, higher BMI, lower SES, lower MEL, and lower proportion of BPW  as the major source of water intake. Fifty four percent of subjects exceeded the AHA daily TASI recommendation, and 47% did not meet the daily TWI recommended by the IOM. Subjects with mothers of lower SES, lower MEL, lower proportion of BPW, higher BMI, and younger age showed a significant higher risk association for such unhealthy beverage pattern intake. There is a remarkably high prevalence of an unhealthy pattern of beverage intake, and specific maternal factors may be implicated as enablers of such behaviors, and also addressable for future interventions