Forage systems included: Old World bluestem (Bothriochloa bladhii) + legumes ) (OWB-legume), native grass-mix (native), alfalfa + tall wheatgrass (Thinopyrum ponticum) , and annual grass-mix  pastures on a clay loam soil; and native, teff (Eragrostis tef), OWB-grazed, and OWB-ungrazed pastures on a sandy clay loam soil. The perennial OWB-legume and native pastures had increased soil organic matter (SOM) and reduced bulk density (ρb), improving sk, soil water thresholds, λ, and tD, compared to annual teff and alfalfa-TW (P < 0.05). Soil λ, but not tD, increased with increasing θv. Grazed pastures decreased sk and water retention compared to other treatments (P < 0.05), yet did not affect λ and tD (P > 0.05), likely due to higher ρb and contact between particles. Greater λ and tD at saturation and PWP in perennial versus annual pastures may be attributed to differing SOM and ρb, and some a priori differences in soil texture. Overall, our results suggest that perennial pasture systems are more beneficial than annual systems for soil water storage and heat movement in semiarid regions.Semiarid pasture management strategies can affect soil hydraulic and thermal properties that determine water fluxes and storage, and heat flow in unsaturated soils. We evaluated long-term (>10 years) perennial and annual semiarid pasture system effects on saturated hydraulic conductivity ( Broadlyn extent . These s content ,4. These content ,6, root  content , soil fa content , shrinki content , freezin content ,11, and  content , wheel-t content , and ani content .λ), volumetric heat capacity (vC), and thermal diffusivity (tD), can be affected by soil texture, soil structure, mineralogical composition, aggregate size, ρb, degree of compaction, soil porosity, soil water content, and SOM [λ, vC, and tD) influence crop production systems by affecting seed germination, seedling emergence, water and nutrient uptakes by plant roots, root growth, and crop growth [Similarly, soil thermal properties influence the partition of energy at the soil surface and control the soil thermal regime, thereby affecting coupled liquid water, water vapor, and heat transport in unsaturated soils ,16,17. T and SOM ,19. Thesp growth ,21.Grassland pastures are increasingly valued for their roles in conserving water and providing sustainable and economically viable alternatives to irrigated crop monocultures for the High Plains agriculture in the USA . Although) of 0 cm) to h of −50 cm [Several studies have indicated ecological linkages between aboveground and belowground processes, as affected by plant community composition , especially in C3 and C4 species ,29. For f −50 cm . SchwartStudies on the effect of grassland management on soil physical properties have typically focused on variations in hydraulic conductivities due to water management, tillage, cropping systems, cover crops, soil amendments, and grazing. However, information on long-term pasture management effects on soil water retention characteristics for various annual and perennial pasture systems is limited. There is also limited information on the effect of annual and perennial pasture systems on soil water retention characteristics and their relationships to soil thermal properties. This lack of information is equally true for semiarid pasture systems with clayey soils. We considered that enhanced soil water and thermal regimes in the upper 20 cm soils were associated with improving soil physical, hydraulic, and thermal properties influenced by long-term perennial and annual pasture management practices. The objectives were to evaluate and analyze: (1) the effect of semiarid perennial and annual pasture management practices on soil hydraulic and water retention characteristics in two pasture soils with contrasting clay contents and (2) the effect of these pasture systems on soil thermal properties as a function of soil water contents given by soil water retention curves (SWRCs).The study was conducted at the Texas Tech University’s Forage-Livestock Research Laboratory Farm, New Deal, located 24 km northeast of Lubbock, TX, USA . The study site represented unsaturated soils that have undergone semiarid pasture management since 1997 . This inGossypium hirsutum L.) system. Treatments were only compared within, rather than between, these selected pasture areas to avoid confounding effects of time since the establishment and management history. The differences in ρb, soil texture, and SOM between these pastures were also emphasized in previous studies [The experimental site for this study had two major pasture areas, of which the east pasture area (12.3 ha) , top was studies ,36. OverEragrostis tef (Zucc.) Trotter), native grass-mix  Engelm), blue grama (Bouteloua gracilis (Willd. Ex Kunth) Lag. Ex Griffths), sideoats grama (Bouteloua curtipendula (Michx.) Torr), and green sprangletop (Leptochloa dubia (Kunth.) Nees)), or Old-World bluestem cv. WW-B.Dahl  T. Blake). The east pasture area had pasture systems containing either mixtures of OWB + alfalfa (Medicago sativa L.) + yellow sweetclover , alfalfa + tall wheatgrass  Beauv.), native, or teff (Eragrostis tef) (only in 2016). Since one of the replicate pastures in the east area had teff only in 2016 and was under different annual crops in previous years, such as sorghum–sudangrass (Sorghum bicolor (L.) Moench), wheat (Triticum aestivum L.), cereal rye , or left fallow  during a severe drought in 2011 and 2012, the pasture was treated as annual. The native pastures in the east area did not have buffalograss. The grazing versus ungrazed (haying only) effect on soil hydraulic and thermal properties were evaluated for OWB pastures in the west area.During the study period (2016–2017), the west pasture area had pasture systems that contained either teff  at different soil depths down to 20 cm . Each aluminum liner with a core sample was removed from the core sampler, labeled, and sealed immediately with polyethylene end caps at both ends of the liner to prevent both soil loss and evaporation. A total of 396 soil cores (11 pastures × 3 replicates × 3 locations × 4 soil depths) were collected from the experimental site each year. All soil samples were transferred from the experimental site to the Soil Physics Laboratory at Texas Tech University. Bulk samples were air-dried, crushed, and sieved to pass a 2 mm screen before further analysis of soil properties. Bulk and core samples were stored in the refrigerator at 4 °C before analysis.g) of soil samples was determined from a 55 g sub-sample taken from fresh bulk samples. A pre-weighed aluminum foil was used to place the soil in a drying oven  at 105 °C for at least 24 h until a constant dry weight was recorded. The θg (g g−1) was determined by dividing the weight of water by the weight of dry soil. About 50 g of air-dried sample (<2 mm) was used to conduct particle size analysis using the sedimentation (hydrometer) method [ρb (g cm−3) by the core method [ρb was calculated as the ratio of the mass of dry soil solids (after oven drying at 105 °C for 24 h) to the bulk volume (volume of solids plus soil pores). The SOM content of soil samples was determined using the loss-on-ignition procedure that estimates SOM based on the gravimetric weight change associated with the high-temperature oxidation of organic matter [−1) that was present in the sample.The gravimetric water content (θ) method  with thee method , in whicc matter . About 1ks (cm d−1) were made on soil core samples (8 cm in diameter and 5 cm in height) by using both constant head and falling head methods [ks measurement, air-dried bulk soil samples from 0–10 and 10–20 cm depths with known ρb values were repacked into 8 × 5 cm sampling cylinders. The measurement of ks was repeated three times for each core sample. In addition, a limited number of core samples using instrument-specific 8 × 5 cm cylinders were also collected from 0–10 and 10–20 cm soil depths to verify ks measurements of saturated repacked soil cores with saturated intact cores using the KSAT instrument. Both constant head and falling head methods, which determine soil ks using Darcy’s equation, are represented by the following governing equations (Equations (1) and (2), respectively).ks is the saturated hydraulic conductivity (cm s−1), V (cm3) is the volume of water flowing through the cross-sectional area A (cm2) of the soil sample per unit time t (s), Q is the steady-state flow rate from Mariotte flask (cm3 s−1), H is the hydraulic head difference between the water inlet and outlet level (cm), L is the length of the soil sample (cm), a is the cross-sectional area of the burette (cm2), H0 and tH are the initial and final hydraulic head differences in the burette (cm), and tt is the time taken for the pressure head to drop from the initial to final pressure head in the burette (s).Measurements of  methods  with a Kh): 0 (at saturation), −100, −330, −500, −2000, −3000, −5000, −10,000, and −15,000 cm. Core samples were saturated from the bottom, weighed, and then placed on a saturated ceramic pressure plate in the pressure plate extractor. A predetermined h was applied for 36 to 48 h. Once the cores were equilibrated to applied h, cores were removed from the extractor and weighed. The same procedure was repeated with successively lower (more negative) h . The ceramic plates corresponding to higher h (saturation to −3000 cm) and lower h  were used to determine SWRCs (soil water retention curves) in the wet and dry soil water ranges, respectively. At the end of the experiment, gravimetric (θg) and volumetric (θv) water contents (cm3 cm−3) of the core samples were estimated from known ρb (g cm−3) of the core samples, dry weight of the core samples, and weight of the core samples at the end of each pressure head (h) experiment. The θv determined at each applied h was averaged over three replicate soil cores. The θv values were plotted against the corresponding h values to obtain functional relationships between θv and h of the soil samples, i.e., SWRCs for all treatments in the east and west pasture areas.Soil water retention characteristics were determined using the pressure chamber method  for all r and θs are the residual and saturated θv, respectively (cm3 cm−3); h is the pressure head (cm); and αv  and n  are fitting shape parameters of the SWRC. Soil water thresholds, necessary for water management in agricultural systems, were obtained from measured SWRCs at different soil depths for all treatments in each pasture area. Soil water for h > −15,000 cm was considered unavailable to plants. The θs  provided a measure of the total porosity of soil samples. Plant available water (PAW) was determined as the difference between field capacity water content , which is the upper limit of PAW, and permanent wilting point  when the plant can no longer extract water from the soil.To quantify water flow parameters in unsaturated soils under long-term pasture management practices in both pasture areas, the van Genuchten’s SWRC model , which hλ (W m−1 K−1), vC (MJ m−3 K−1), and tD(mm2 s−1) were made on 5 × 5 cm core samples taken at soil depths of 0–5, 5–10, 10–15, and 15–20 cm using the KD2 Pro Thermal Properties Analyzer . The λ, vC, and tD values were measured simultaneously during the SWRC measurements at applied pressure heads (h) of 0 (at saturation), −100, −330, −500, −2000, −3000, −5000, −10,000, and −15,000 cm. The KD2 pro instrument, equipped with a dual-needle sensor , works on principles of the transient line heat source analysis [λ, vC, and tD of core samples using a Delrin verification block supplied by the manufacturer.For all treatments in each pasture area, measurements of analysis ,46. The λ, vC, and tD. During the measurement of λ, vC, and tD, the KD2 pro instrument applied heat for a set of heating times (ht) to one of the needles that contained a heating element. The heat was then transferred into the soil between two needles, and the temperature was measured in the measuring needle during the heating and the cooling period following heating. The resulting data were fitted to the following equations (Equations (4) and (5)) using a nonlinear least-squares procedure [T is the temperature rise at the measuring needle; T0 is the temperature at the start of the measurement; T is the measured temperature; b0, b1, and b2 are the fitting parameters; q is the heat input at the heated needle (W m−1); iE is the exponential integral that is approximated using polynomials [t is time (s); and ht is the heating time (s). Equation (4) was applied for the first ht when the heat was on, while Equation (5) was applied when the heat was off. The λ and tD were then computed from Equations (6) and (7), respectively, while vC was given as the ratio of λ and tD.r is the distance from the heated needle to the measuring needle.During SWRC measurements, after the soil water equilibrium had been reached at each of the applied pressure heads, the core samples were removed from the pressure extractor, and the dual-needle sensor was inserted at the full depth of the needles into each core sample to determine rocedure .(4)ΔT=bynomials ; t is tiMeasured data were analyzed separately for the east and west pasture areas. The generalized linear mixed model (PROC GLIMMIX) procedure in SAS version 9.4  was usedP > 0.05, P < 0.01), while treatments did not affect SOM in the west in 2017 (P > 0.05). In the east pasture area, OWB-legume had the greatest SOM compared to alfalfa-TW, native, and annual (P < 0.01), whereas the annual had the lowest SOM (P < 0.01). OWB-grazed and OWB-ungrazed had consistently greater SOM than teff in 2016 (P < 0.05), while the native was intermediate between OWB and teff. Grazing did not affect SOM in the west OWB pasture (P > 0.05).Sand, silt, and clay contents in unsaturated soils were not affected by management practices in both east and west pasture areas  than in the 5–20 cm depth (P > 0.05). In the east area, ρb in the 0–5 cm depth differed among pasture treatments (P < 0.05), whereas treatment had no effect in the deeper soil depths (P > 0.05). OWB-legume pasture had lower ρb than alfalfa-TW and annual in the upper 0–5 cm, while native had intermediate ρb values between OWB-legume and alfalfa-TW and annual pastures. On the contrary, ρb in the 0–5 cm was not affected by treatments in the west area (P > 0.05). Nonetheless, the treatment effect on ρb was observed for 5–10 and 10–15 cm depths (P < 0.05). Ungrazed OWB showed lower ρb than OWB-grazed and teff (P < 0.05). As in the east pasture, natives had an intermediate ρb value between OWB and teff in the west area. Grazing increased ρb in OWB pasture by 10% over OWB-ungrazed within the 5 to 15 cm soil depth in the west area, likely owing to animal trampling as SOM content was the same in both OWB-grazed and OWB-ungrazed pastures  values were 61 and 57 cm d−1 at depths of 0–10 and 10–20 cm, respectively, in the west pasture area during 2016–2017. The respective ks values for these depths in the east area were greater, i.e., 113 and 103 cm d−1, even though the east pasture soils had higher clay contents. Notably, the east pasture area tended to have lower ρb values than the west  and characteristics of pores in soils have been known to impact ks. In addition, changes in vegetation species could be a driving factor influencing hydraulic properties by altering total porosity, non-capillary porosity, and macro water-stable aggregates [ks through root distribution and morphological characteristics, such as root biomass and distribution , while it was significant for the 10–20 cm depth in the east and both 0–10 and 10–20 cm depths in the west area (P < 0.05). In the east area, native pasture had higher ks than annual and alfalfa-TW in the 10–20 cm depth (P < 0.05). For the 0–10 cm depth in the west area, OWB-ungrazed had higher ks than teff, and teff had the lowest ks (P < 0.05). Native and OWB-grazed pastures had intermediate ks values between OWB-ungrazed and teff. In the west area, ks in the 10–20 cm was in the order of native > OWB-ungrazed > OWB-grazed > teff. Saturated hydraulic conductivity (ks) was associated with ρb  and SOM  (data not shown) as it was greater for perennial pastures with low ρb and high SOM than annual. Our result was similar to previous studies, in which lower ρb was aligned with higher ks  for all treatments  between 0.010 and 0.038 cm3 cm−3 and 0.013 and 0.025 cm3 cm−3 (data not shown) for the east and west pasture areas, respectively. The measured and fitted SWRCs for different pasture soils suggested that soils with greater SOM and ks retained more water at or near saturation  and released less water when the h increased , except for native and OWB-ungrazed pastures in the west area. Nonetheless, the capillary region  of the SWRC was steeper with an increase in h for soils with greater SOM  in the wet and dry soil water ranges of SWRCs. Fitted van Genuchten parameter values within the 0–20 cm soil depth for all treatments in both pasture areas are listed in r values among different treatments in both pasture areas. The residual water content (θr) represents θv at the lowest h, at which water is retained in soil micropores [r is often related to the amount and state of water adsorption in the mineral fractions of soil [vα, which determines the air entry h required to initiate the release of water and air begins to enter the soil pores during drainage when h exceeds air entry h, and especially parameter n, which describes the rate of water desorption, varied among pasture treatments within the 0–20 cm soil depths , with greater FC (0.38 to 0.44 cm3 cm−3) and PAW (0.17 to 0.23 cm3 cm−3). Generally, the east area had more water holding capacity (and PAW) than the west, which could be explained by the higher clay contents and SOM of the east pasture soils  at different soil depths for each treatment, are shown in re soils .sk and root characteristics  was not clarified in our study, root parameters might have had effects on SWRC or sk under different pasture systems [v at saturation  in the 0–5 cm depth was greater for OWB-legume than alfalfa-TW and annual (P < 0.02), suggesting that OWB-legume treatment provided the relatively good topsoil structure.It is worth noting that the type of OWB species used in this experiment  has been promoted for its longer vegetative period than other OWB species, which favors greater production of above- and belowground biomass . Althoug systems ,72. In t3 cm−3 more water content in the 0–5 cm between h = 0 cm (at saturation) and h = −100 cm. Alfalfa-TW had the lowest total porosity in the 5–10 cm, where there was a parity among OWB-legume, native, and annual treatments (P < 0.05)  . In gene−2000 cm . Total pv at saturation than OWB and native perennial pastures (P < 0.05) in the upper 0–5 cm depth , but after that, native and teff released more water at an increasing rate with increasing  h when compared with OWB pastures. OWB-grazed delayed release of water after h of −1000 cm. OWB-ungrazed retained water more tightly, close to the grazed one in the 10–15 cm under h of −2000 cm or lower. At the 15–20 cm depth, native and OWB-ungrazed had improved soil water retention characteristics at h less than −100 cm than teff and OWB-grazed. Nevertheless, native and OWB-ungrazed released water under increasing h, while the soil impacted by grazing (OWB-grazed) held more water under the increased  h (P < 0.05) in the 15–20 cm, which was suggested by higher ρb at this 15–20 cm soil depth.In the west pasture area, teff provided lower θcm depth . As showative) h . Grazingv at saturation were observed for FC soil water threshold  (P < 0.05) in the 0–5 cm. Annual pasture had an intermediate FC value between native and alfalfa-TW. Native in the 5–10 cm and OWB-legume in the 10–20 cm had greater FC than alfalfa-TW (P < 0.05). Both native and annual treatments provided mostly similar intermediate FC values between OWB-legume and alfalfa-TW. In the west area, the effects of pasture management on FC soil water thresholds were not significant for 0–5 and 5–10 cm depths (P > 0.05). However, the effects of pasture management on FC for 10–15 and 15–20 cm depths were significant (P < 0.05). OWB-grazed treatment provided lower FC than OWB-ungrazed and native at depths of 10–20 cm and 15–20 cm, respectively (P < 0.05). As mentioned earlier, although soils in the west area had higher sand contents, higher ρb under OWB-grazed likely diminished the volume of large pores, affecting soil water retention at field capacity in denser soils.Similar effects of pasture management to those discussed for θapacity) . In the P > 0.05). In the west area, OWB-grazed showed greater PWP than OWB-ungrazed and native in the upper 0–5 cm (P < 0.05). OWB-grazed showed higher PWP compared to native pasture (P < 0.05) in the 0–5 cm depth (P < 0.05). Notably, grazing in the west area did not affect θv at PWP at soil depths of 5–10 cm and 15–20 cm (P > 0.05).As compared to FC soil water thresholds, an opposite trend was observed for PWP soil water threshold values under different treatments , where p3 cm−3 and 0.095 to 0.190 cm3 cm−3 for the east and west pasture areas, respectively, within the 0–20 cm depth (P < 0.05). However, for the 5–10 cm depth, native pasture showed greater PAW than OWB-legume (P < 0.05). Alfalfa-TW treatment had PAW (0.120 cm3 cm−3), almost half of that in the OWB-legume treatment (0.229 cm3 cm−3). In the west pasture area, native and OWB-ungrazed had greater PAW than OWB-grazed in all soil depths (P < 0.05).Following the differences observed in FC and PWP soil water thresholds at different soil depths, PAW ranged from 0.112 to 0.233 cmcm depth . The easλ) and diffusivity (tD) as a function of volumetric water contents (θv), which correspond to applied h of 0 (at saturation),−100, −330, −500, −2000, −3000, −5000, −10,000, and −15,000 cm, are shown in λ increased with increasing θv in both pasture areas. The increase in λ values with increasing θv agreed with previous studies . As θv increases towards saturation, the replacement of air with water continues to enhance heat conduction through the mixture because more water gathers around the contact points and forms water bridges  between solid particles [λ increased with increasing θv, there were no discernible changes in tD with increasing θv for all treatments in both pasture areas. The reason could be explained by very little relative changes in the λ and vC values of pasture soils observed in our study. The increase in λ with increasing θv was lower than the increase in vC. On average, vC increased by 13 and 7% when θv increased from PWP (2.33 and 2.35 MJ m−3 K−1) to saturation (2.63 and 2.51 MJ m−3 K−1) (data not shown) for the east and west pasture areas, respectively. This also suggested that while higher λ values ensured faster soil temperature recovery, higher vC values of pasture soils likely provided larger heat storage in the upper 0–20 cm depth.Soil thermal conductivity . Soil λarticles ,77. NotatD values (as a function of θv) varied between 0.39 and 0.80 mm2 s−1 and between 0.48 and 0.76 mm2 s−1 within the 0–20 cm soil depth in the east and west pasture areas, respectively. Soil tD data observed in our study were within the desirable ranges reported in previous studies [λ values in our study were also within and close to the desirable λ ranges that facilitated greater heat movement through the soil. For instance, Ghuman and Lal (1985) [λ ranged from 0.37 to 1.42 W m−1 K−1 at θv of 0.02 to 0.16 cm3 cm−3 for sandy loam, 0.35 to 3.34 W m−1 K−1 at θv of 0.02 to 0.46 cm3 cm−3 for sandy clay loam, and 0.39 to 1.15 W m−1 K−1 at θv of 0.10 to 0.52 cm3 cm−3 for clay soil, respectively.As shown in  studies ,78,79, sλ values for each pasture area showed a sudden increase in λ when θv increased from 0.20 to 0.25 cm3 cm−3, after which it increased slowly with further increases in θv up to saturation (λ of 2.03 W m−1 K−1 was observed at saturation in the 5–10 cm depth under the native pasture system. The regression between λ and θv (λ than OWB-legume at both saturation and PWP in the upper 0–5 cm depth (P < 0.05), while λ value of annual pasture soils was intermediate. Except for alfalfa-TW, the differences in tD at saturation among native, alfalfa-TW, and OWB-legume followed a similar pattern of λ values for the same soil depth. For the 5–20 cm depth, tD at saturation did not vary between treatments. At PWP, native had greater tD than OWB-legume and alfalfa-TW in the 0–5 cm, whereas, in the 5–10 cm depth, OWB-legume and annual had greater tD than alfalfa-TW (P < 0.05).The depthwise comparison of turation . The maxλ and θv  showed sv (a function of pressure heads) in the west pasture soils . ,83. v  [tD by changing contact areas, and by altering water storage [λ [As discussed in λ ,84. Orgare space ,86, doesh   observedc (2020) , in whicorage [λ .λ and tD values among different treatments were evaluated by three soil water thresholds, namely, very low θv (at PWP), intermediate θv (at FC), and high θv (at saturation). Soil λ and tD at saturation and PWP in different soil depths are presented in λ and tD values in the upper 0–10 cm depth at saturation and PWP were affected by pasture treatments (P < 0.05), whereas λ and tD in the 10–20 cm depth were not affected by treatments (P > 0.05). Soil λ and tD at FC were not affected by pasture treatments (P > 0.05) (data not shown). Soil λ and tD at FC were 1.03 and 1.24 W m−1 K−1 and 0.55 and 0.66 mm2 s−1 in the east and west areas, respectively, when averaged across years and soil depths. Soil λ and tD values at FC were 1.31, 1.27, 1.37, and 1.31 W m−1 K−1 and 0.54, 0.53, 0.58, and 0.57 mm2 s−1 for native, OWB-legume, annual, and alfalfa-TW, respectively, in the east pasture area, while in the west area, λ and tD at FC were 1.51, 1.54, 1.55, and 1.60 W m−1 K−1 and 0.65, 0.67, 0.68, and 0.70 mm2 s−1 for native, OWB-ungrazed, OWB-grazed, and teff, respectively. Neither treatment × depth nor year × treatment interaction was significant for both λ and tD in both pasture areas (P > 0.05).Based on SWRCs data , the varλ had no definitive trend at saturation. For example, native showed greater λ than OWB pastures in the 0–5 cm depth, whereas teff and OWB-grazed had greater λ at saturation than native in the 5–10 cm (P < 0.01) . Similarly, tD did not differ among treatments at saturation for all depths (P > 0.05). At PWP, only the upper 0–5 cm depth showed differences in tD, where native had greater tD than OWB-ungrazed (P < 0.01). Teff and OWB-grazed had intermediate tD values between native and OWB-ungrazed. For the 5–20 cm depth, tD did not vary among treatments at PWP (P > 0.05). Overall, greater tD within the upper 0–20 cm soil depths in the west pasture area compared to the east pasture area  . Soil λ ure area  likely aextremes , especiaρb in the upper 0–20 cm soil depth significantly increased sk compared to annual  pasture systems (P < 0.05). The perennial pasture systems also improved soil water thresholds  and thermal properties (λ and tD) than annual pasture systems (P < 0.05), underscoring the benefits of permanent, perennial cover on soil water storage in semiarid environments. Native and OWB-legume pastures, especially in the clay loam east pasture, showed a similar increase in sk and soil water retention characteristics, owing to their greater SOM contents but lower ρb compared to annual teff and alfalfa-TW (P < 0.05). Soil λ increased with increasing θv (a function of pressure heads); however, there were no discernible changes in tD with increasing θv since the increase in vC resulted in very little relative changes in λ and tD of pasture soils across the range of θv given by SWRCs. Differences in greater λ and tD with differences in soil water thresholds between perennial and annual pastures, especially at saturation and PWP, may be attributed to differences in SOM and ρb, in addition to a priori differences in soil texture. Grazed pastures decreased sk and soil water retention compared to other treatments (P < 0.05), yet did not affect λ and tD (P > 0.05), likely due to higher soil ρb and contact areas between particles. Significantly lower soil water retention, sk, λ, and tD in annual and teff pastures compared to perennial systems (P < 0.05) were attributed to annual disturbance, including tillage practices. Overall, our results suggest that management practices involving continued soil disturbance, including annual forage crops and tillage-based row-crop agriculture, are less beneficial to water storage and heat movement in unsaturated soils than perennial pasture systems, especially in semiarid regions.The perennial OWB-legume and native pasture systems that contributed to the build-up of SOM and reduced


Data on manifest refraction, corrected distance visual acuity, dominant eye, presbyopic addition, intraocular pressure, and anterior segment biometric parameters were collected. The visual outcomes and binocular balance at 0.4 m, 0.8 m, and 5 m were documented.p = 0.125), respectively. Binocular visual acuity (logmar) for 0.4 m, 0.8 m, and 5 m were −0.03 ± 0.05, −0.03 ± 0.02, and 0.10 ± 0.03 for the ICL V4c group, and −0.02 ± 0.09, −0.01 ± 0.02, and 0.06 ± 0.04 for the FS-LASIK group, respectively. The proportions of all patients with imbalanced vision at 0.4 m, 0.8 m, and 5 m distances were 68.89, 71.11, and 82.22%, respectively . There were significant differences in refraction between the balanced and imbalanced vision for patients at 0.4 m distance , 0.8 m distance , and 5 m distance .The safety index for the ICL V4c and FS-LASIK groups were 1.24 ± 0.27 and 1.04 ± 0.20 (ICL V4c implantation and FS-LASIK monovision treatment demonstrated good long-term safety and binocular visual acuity at various distances. After the procedure, the imbalanced patients’ vision is primarily related to the age-related presbyopia and anisometropia progression caused by the monovision design. Effective cooperation between the eyes is crucial for obtaining high-quality images. A binocular imbalance may represent inhibitory binocular interactions by determining the signal strength inequality between the eyes . It refein situ Keratomileusis (FS-LASIK). Presbyopia progression may continue postoperatively. Furthermore, uneven refractive errors caused by monovision and age-related neural processing deficits may affect binocular balance. This could compromise patients’ visual function and experience, highlighting the need for careful consideration of the potential benefits and drawbacks of this intervention  <2.8 mm.This study enrolled 90 eyes in 45 patients  An RT-5100 phoropter  was used to measure spherical equivalent (SE), corrected distance visual acuity (CDVA), uncorrected-distance visual acuity (UDVA), and presbyopic add power (ADD). The safety index (SI) was defined as the postoperative CDVA over the preoperative CDVA, and the efficacy index (EI) was defined as the postoperative UDVA over the preoperative CDVA. D-eyes and nD-eyes were determined using the card-hole method. ADD was measured using the Fusion Cross-Cylinder (FCC) method at a 33 cm distance with optimal distance visual acuity correction. (2) Intraocular pressure (IOP) was measured using a Canon Full Auto Tonometer TX-F . (3) Corneal thickness (CT), anterior chamber volume (ACV), ACD, anterior chamber angle (ACA), and white-to-white ratio (WTW) were measured using Pentacam HR . (4) Endothelial cell density (ECD) was measured using SP-2000P . (5) White-to-white (WTW) measurements were performed using IOL Master 700 . (6) Axial length (AL) was measured using IOL Master 500 . Slit lamp and fundus examinations were conducted following pupillary dilation to evaluate lens transparency and exclude fundus lesions.2.3.The D-eyes were targeted for −0.34 to 0.02 D and −1.25 to 0.25 D in the ICL V4c and FS-LASIK groups, respectively. The targeted nD-eyes were around −2.435 to −0.27 D and −2.75 to −0.25 D in the two groups, respectively, according to each patient’s presbyopic add power. In those with planned residual myopic diopters, the target refraction was on trial in-frame glasses preoperatively and was accepted by the patients.2.4.The choice of operation was confirmed by surgical indications after adequate communication with patients. For patients who met the two surgical indications at the same time, they could choose by themselves with the knowledge of the two surgeries. The same surgeon (XZ) performed all surgical procedures. The patients were administered antibiotic eye drops four times daily for 3 days before surgery. In FS-LASIK, a 500 kHz VisuMax femtosecond laser system  with a pulse energy of 130 nJ was used for flap creation, followed by a MEL 90 excimer laser (Carl Zeiss Meditec) for stromal ablation, with a pulse energy of 185 nJ. The flap diameter and thickness were 7.5 mm and 100 μm, respectively, with standard 90° hinges and 90° side cut angles. The planned optic zone of 6.45 ± 0.20 mm (ranging from 6.00 to 6.80). After FS-LASIK, a soft contact lens was worn and removed on 1 day postoperatively. Topical levofloxacin, 0.1% fluorometholone solution, and non-preserved artificial tears were administered after FS-LASIK. The detailed steps have been previously described .The STAAR Surgical online calculator (Version 3.0)2.5.The patients were followed up for 4 years, with an average of 48.73 ± 14.65 months. At the last follow-up, the CDVA and UDVA (logMAR) of the D eyes, nD eyes, and both eyes at 0.4 m, 0.8 m, and 5 m were recorded. Measurements were performed using two trial frames and two tumbling E charts  for the monocular and binocular VA at the distance of 0.4 m and 0.8 m, respectively. A phoropter was used for monocular and binocular distant VA at 5 m.2.6.Binocular balance was assessed using a modified version of the dichoptic procedure proposed by 2 and a refresh rate of 120HZ). All patients wore 3D polarized glasses to perform the BI tests at a distance of 0.4 m, 0.8 m, and 5 m; each eye was presented with either horizontal or vertical stripes individually, with 100% contrast (The stimulus image was presented on a three dimension (3D) gamma-corrected monitor . Results are expressed as mean ± standard deviation. The normality of the data was checked using the Kolmogorov–Smirnov test. Repeated ANOVA was used to compare the pre-and post-treatment and D- and nD eyes normally distributed data, while the paired t-test was used to compare the normally distributed data of the D and nD eyes at the last follow-up, and the Wilcoxon signed-rank test was used for non-normally distributed data. A generalized estimating equation was used to determine the correlation between binocular balance parameters  and refraction parameters . Differences were considered statistically significant at 3.All surgeries and examinations were performed, and no complications, such as cataracts or high intraocular pressure, occurred in any eye throughout the follow-up period. The data loss rate for all the types was <5%.3.1.p = 0.125), respectively. Their efficacy indices were 0.77 ± 0.29 and 0.66 ± 0.34 (p < 0.001), respectively (2 (17.14 ± 14.88% and 3.59 ± 3.11% per year). The vault of all eyes after ICL V4c implantation was within the 150 ~ 850 μm range.At the last follow-up, the safety index of the ICL V4c and FS-LASIK groups were 1.24 ± 0.27 and 1.04 ± 0.20  of the ICL V4c group were −0.03 ± 0.05 (0.4 m), −0.03 ± 0.02 (0.8 m), and 0.10 ± 0.03 (5 m). The corresponding figures were −0.02 ± 0.09, −0.01 ± 0.02, and 0.06 ± 0.04 in the FS-LASIK group. The two groups had no significant difference . The binocular balance level was related to the monovision design and SE of the non-dominant eye at the last follow-up     (The SE of non-dominant eye (−1.14 ± 0.17D) for patients with binocular balance at 0.4 m significantly differs from that with binocular imbalance (−1.47 ± 0.13D) (< 0.001) . The ran= 0.027) , visual = 0.027) , and age= 0.030) .3.4.2.B = 2.72, p = 0.041). Additionally, there was a significant difference in binocular visual acuity at 0.8 m between the binocular balanced patients (−0.01 ± 0.03) and the imbalanced (−0.02 ± 0.01D) at 0.8 m .There was a significant difference in preoperative ADD between the binocular balanced (0.90 ± 0.17D) and the imbalanced (1.05 ± 0.11D) patients at 0.8 m . Additionally, there was a significant difference in target SE of non-dominant eye between the binocular balanced patients (−0.78 ± 0.19D) and the imbalanced (−1.17 ± 0.13D) at 5.0 m  and the real refraction of the dominant eye . The range of balance of the dominant eye may be correlated with age  and the visual acuity of the dominant eye at 5.0 m   and the imbalanced (−1.42 ± 0.11D) at 5 m (= 0.036) .4.This study elucidated the safety index of the two surgical procedures after 4 years postoperatively, which was 1.24 ± 0.27 and 1.04 ± 0.20, for the ICL V4c and FS-LASIK groups, respectively, while the efficacy index was 0.77 ± 0.29 and 0.66 ± 0.34, respectively. The efficacy index was lower owing to intentional undercorrection in the non-dominant eye than in conventional studies. However, considering good binocular visual acuity at near-to-far distances, monovision surgery using ICL V4c or FS-LASIK has good long-term safety and efficacy in myopic patients with presbyopia. With the increasing number of myopic patients and the aging of the population, the safety and efficacy of refractive surgery in people aged ≥40 years have received widespread attention . The safAnisometropia between the dominant and non-dominant eyes caused by monovision may affect the visual function, which may affect their visual experience. A study has shown that anisometropia may lead to binocular imbalance and accommodative difficulties . This imThis study showed that at different distances, the balanced range of the dominant eye was related to the visual acuity or SE of the non-dominant eye and also showed a correlation with age at far and near distances. The correlation of visual acuity was that the closer the binocular visual acuity, the higher the balance range. The correlation between the dominant eye’s balance range and the non-dominant eye’s SE showed the opposite result. Thus, the higher the degree of under-correction of the non-dominant eye at near distances, the lower the degree of under-correction at far distances, and the higher the balance level of the dominant eye. This contrasts the extreme relationship between this parameter and eye balance. Previous studies have suggested that an increase in monocular blur may increase the stereo threshold at high spatial frequencies  and thatPeople commonly experience binocular imbalance, with 65.9 and 62.89% experiencing low-and high-temporal-frequency stimuli, respectively . The proResearch conducted among myopic adolescents aged 6–18 years showed that an increase in binocular imbalance does not necessarily imply poorer SE or VA. Instead, an increase in anisometropia significantly correlated with an increase in binocular imbalance, and the dominant eye tended to have a more negative SE than the non-dominant eye . This stThis study had certain limitations. First, the preoperative binocular balance was not compared. However, with the advancement and optimization of detection methods for binocular balance, future studies can better evaluate the changes before and after monovision surgery. The short-term plasticity of visual perception training can also be added to increase data variability before and after training, which can provide valuable clinical support. Second, tests for stereo vision function were not applied in this study, and future research is needed as binocular imbalance and stereo vision are two different aspects of binocular interactions. Previous studies have shown that subjects with good binocular balance tend to have better stereo vision. Therefore, studying different aspects of binocular relationships, such as stereo vision, in this population will contribute to further evaluating the clinical effects of monovision surgery. Third, the binocular imbalance was not measured during different working hours, and transient binocular imbalance is likely a temporary physiological phenomenon . As timeIn conclusion, the ICL V4c implantation and FS-LASIK monovision treatment demonstrated good long-term safety and binocular visual acuity at various distances. The imbalanced patients’ vision is primarily related to the age-related presbyopia and anisometropia progression caused by the monovision design. High levels of anisometropia may required careful consideration from the aspect of biocular balance.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.This study followed the tenets of the Declaration of Helsinki and was approved by the Ethics Committee of Fudan University Eye and ENT Hospital Review Board . Informed consent was obtained from all participants. The patients/participants provided their written informed consent to participate in this study.YY, ZZ, JZ, and XZ: study concept and design. YY, ZZ, LN, WS, XW, and JZ: data collection. YY: data analysis and interpretation. YY and JZ: drafting of the manuscript. LY, JZ, and XZ: critical revision of the manuscript. XZ: supervision. All authors read and approved the final manuscript.This study was supported by National Natural Science Foundation of China (Grant no. 82271119), Shanghai Rising-Star Program (23QA1401000), Healthy Young Talents Project of Shanghai Municipal Health Commission (2022YQ015), Project of Shanghai Science and Technology (Grant nos. 20410710100 and 21Y11909800), and Project of Shanghai Xuhui District Science and Technology (2020-015).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


This study aimed to evaluate salivary pH changes after consuming three types of milk in children aged 3–5 years. The null hypothesis was that no statistically significant difference would be noted in the salivary pH between high-protein, full-fat, and sweetened milk groups at different time points.0) after 5 (t1), 10 (t2), 15 (t3), 30 (t4), and 60 (t5) minutes of milk consumption, using a pH saliva indicator strip.This was a double-blind, pilot randomized controlled crossover trial. 30 Children have undergone three experimental sessions with a 1-week washout period. Each child was given 250 mL of one of the following types of milk: high-protein, full-fat, or sweetened milk. The salivary pH was measured at the baseline (tP < 0.05) and full-fat milk consumption (p < 0.05). However, the initial drop in the salivary pH was found to remain above the critical level. For the high-protein milk group, salivary pH decreased slightly after 5 min but was similar to that at the baseline (p = 0.573). In the high-protein milk group (p < 0.05), the salivary pH was slightly greater than the baseline value at t5.There is a sharp drop in salivary pH after 5 min of sweetened (The study shows an initial suggestion that milk is a non-cariogenic beverage, even when sugar is added. Furthermore, high-protein milk has a protective effect from dental caries. Milk is a liquid produced by mammals to meet the nutritional needs of newborns, and it is considered the primary source of nourishment in proper proportions . In addiHuman saliva consists of more than 99% water and less than 1% organic and inorganic substances. The flow rate of unstimulated saliva is about 0.3–0.4 ml/min , and theThis was a double-blind, pilot randomized controlled crossover trial where each participant would be their comparator. It was conducted for 2 months at Damascus University. Ethical approval was provided by the Institutional Review Board at Damascus University (N7748), and it was conducted in full accordance with the 1964 Helsinki Declaration  and CONShttp://www.gpower. hhu.de/). Effect size f = 0.5996822/α err prob = 0.05/ Power (1-β err prob) = 0.80/ Number of groups = 3/ Total sample size = 30. Effect size was estimated according to the results of a pilot study of five participants, and a sample size of 30 children was sufficient to obtain an effect size of 0.59.Sample size was calculated using G*Power software 3.1.9.4  .Children aged 3–5 years.Children who are definitely positive, or positive according to Frankl’s behavior rating scale.Children with full primary dentition.Taking medications that affect the salivary flow rate.Intolerance to milk protein.Children undergoing orthodontic treatment.33 patients were assessed for eligibility, and 3 were excluded due to not meeting the inclusion criteria. The modified CONSORT flowchart is presented in Fig. 0) then each child was given 250 mL of one of the following types of milk:High-protein milk .Full-fat milk .Sweetened milk .Children have undergone three experimental sessions with a 1-week washout period . The seq1), 10 (t2), 15 (t3), 30 (t4), and 60 (t5) minutes of milk consumption. Salivary pH was measured using a pH saliva indicator strip , which is to Alkarad et al. [The salivary pH was measured after 5 .Full-fat milk .Sweetened milk .The composition of the milk samples according to the manufacturing industry were as follows:High-protein milk (pH = 5.84).Full-fat milk (pH = 5.82).Sweetened milk (pH = 5.51).The starting pH of the milk samples was tested using pH meter . The starting pH was as follows:p < 0.05).Statistical analysis was performed using IBM SPSS software version 24 . Descriptive statistics were calculated and provided as mean, standard deviation, standard error, minimum, and maximum. Kurskal–Wallis test was performed to compare non-paired data, and Wilcoxon signed-rank test was used to compare paired data. Statistical significance was adjusted at 0.05 . The mean age was 4 years . The mean of the dmft score was 4.8 ± 3.9. The salivary pH values were listed as mean, standard deviation (SD), standard error (SE), minimum (Min), and maximum (Max) for each group at different time points , suggesting that the baseline data are well-balanced. However, the Kruskal–Wallis test indicated a significant difference between the study groups at t1, t2, t3, and t4 (p < 0.05) (Table 1 (p < 0.05) and a significant difference between high-protein milk and sweetened milk groups at t1, t2, t3, and t4 (p < 0.05). In addition, there was a statistically significant difference between full-fat milk and sweetened milk groups at t1, t2, and t3 (p < 0.05) . However, there was no significant difference in the salivary pH at t1 (p = 0.573), t2 (p = 0.876), and t3 (p = 0.222) compared to t0. In the full-fat milk group, a significant difference was noted in the salivary pH at t1, t2, and t3 (p < 0.05), but no significant difference was observed at t4 (p = 1.000) and t5 (p = 0.058) compared to t0. In the sweetened milk group, there was a significant difference at the five-time points compared to t0 (p < 0.05) (Table In the high-protein milk group, a significant difference was found in the salivary pH at t5.The Kinetics of salivary pH were presented in Fig. Dental caries is the most common chronic disease in both children and adults. The frequent consumption of dietary free sugars between mealtimes increases the risk of dental caries, especially in schoolchildren . To the Salivary pH is a biomarker for many oral and systemic diseases, such as hypertension, insulin resistance, chronic kidney disease, depression, periodontal diseases, and dental caries. , 28 In a1, t2, and t3 compared to t0. In addition, the salivary pH was slightly greater than the baseline value at t5, and there was a significant difference in the salivary pH at t4 and t5 compared to t0. This result could be explained by the high protein content, which has a high buffering capacity. High protein concentration increases the salivary pH since it increases the flow rate of saliva [In the high-protein milk group, salivary pH decreased slightly after 5 min but was similar to that at the baseline, and there was no significant difference in the salivary pH at tf saliva , 33. As f saliva , suggestf saliva , salivar1, t2, and t3 compared to t0. Subsequently, a gradual recovery was noted within 60 min, and no significant difference was observed at t4 and t5 compared to t0. This could be explained by the fact that milk fat adsorbs to the surface of the enamel and may play a protective effect [For the full-fat milk group, there is an initial drop in salivary pH after 5 min of consumption, and a significant difference was noted in the salivary pH at te effect . Howevere effect  suggestee effect  found th1. This finding is consistent with one reported by Seralurin et al. [The result of the current study suggested that there is a significant difference between high-protein milk and full-fat milk groups at tn et al.  suggesti0. This result is similarly reported by Hirani et al. [In the sweetened milk group, there is a sharp drop in salivary pH after 5 min of consumption. Subsequently, a gradual recovery was noted within 60 min and there was a significant difference at the five-time points compared to ti et al. . Howeveri et al.  study, ti et al. . As a rei et al.  study, si et al.  found thi et al.  reportedi et al.  suggestei et al. . Hence, i et al.  result sThis study has limitations. Cross-over trial designs are subjected to carryover effects, and it is hard to accurately estimate the washout period required. In addition, the effect of each type of milk was only tested once. As milk is not consumed only once by each child. Furthermore, litmus paper is not as accurate as pH meter .Within the limitations of this study, it shows an initial suggestion that milk is a non-cariogenic beverage, even when sugar is added. Furthermore, high-protein milk has a non-cariogenic potential and high buffering capacity. Hence, it has a protective effect from dental caries. Further trials with larger sample sizes are recommended to ascertain results. In addition, it would be appropriate to conduct an experiment where milk is consumed 3 times per day during a period of at least 7 days. Furthermore, it is recommended to use a microelectrode technology, which is more precise to evaluate the salivary pH changes. However, this method was not used due to its unavailability.


Ovarian cancer is the most lethal cancer in gynaecology. Surgery, chemotherapy, and radiotherapy are the most often used cancer-fighting strategies. Post-surgery infection is fairly prevalent, especially among people with insufficient immunity. Zinc oxide nanoparticles (ZnOnps) have amazing biomedical features as anticancer and antibacterial agents.Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were tested as models for gram + ve and gram -ve bacteria. The selected bacteria were subjected to concentrations of 20, 40, 80, and 100 μg/ml.We investigated the behaviour of ZnOnps synthesized by green methods on ovarian cancers using established human ovarian cancer cell lines, besides the antibacterial action toward models of gram + ve and gram -ve bacteria. The cytotoxic effect of ZnOnps was calculated using a Sulforhodamine B (SRB) trial. S. aureus and 25.66 ± 0.33 to 31 ± 0.33 mm for E. coli. ZnOnps antagonistic effect statistically differed with neomycin, cefaclor, and cefadroxil.The synthesized ZnOnps induced 50% inhibitory concentration (IC50) at a concentration of 27.45 μg/ml. The diameter of inhibition ranged between 20.16 ± 0.16 and 27 ± 0.57 mm for S. aureus and E. coli infection.Green synthesis of ZnOnps is easily prepared, low cost, non-toxic, and eco-friendly. Their cytotoxic action on SKOV3 cells and their antibacterial characteristics pave the way to be an alternative therapy for ovarian cancer and  Cancer is the world’s second cause of mortality , 2. OvarOne of the biggest challenges to modern public health is antimicrobial resistance (AMR). It has been promoted by increased exposure and the misuse of antibiotics , 8. AccoAs a result, we have had to explore designing safe, effective, and environmentally friendly antimicrobial agents , 11. SecStaphylococcus aureus (S. aureus) infection varies between 1 to 3 cases per 10,000 people [S. aureus is a prevalent human pathogen accountable for local infections. S. aureus has proved to be resistant to commonly used antimicrobial agents, e.g., penicillin, methicillin, tetracycline, erythromycin, and vancomycin [The annual incidence of 0 people . S. aurencomycin . Most pancomycin . Microbincomycin , alternancomycin , antibioncomycin .Infections are one of the most frequent complications in patients with cancer . A patieThe most frequent cause of infections in cancer patients is bacteria. Antibiotic failure increases the risk of sepsis, sepsis-related death, and sepsis-related care costs in cancer patients. According to a recent survey conducted in the United Kingdom, 46% of oncologists are concerned that drug-resistant infections may make chemotherapy problematic as a cancer treatment .Green synthesis is defined as using environmentally compatible materials such as bacteria, fungi, and plants to synthesize nanoparticles . Zinc oxThe mode of action of ZnOnps as an anticancer and antibacterial can be explained by metal oxide nanoparticles interacting with bacteria or cells, producing reactive oxygen species (ROS). By assaulting the cell membrane and then attaching to the -SH group of cellular enzymes, the respiratory chain is impacted by the metal ions the nanoparticles create, and certain enzymes are blocked as a result. Therefore, singlet oxygen, hydroxyl radicals, hydrogen peroxide, superoxide anions, and other ROS are created and start to build up. ROS can harm the proteins and DNA found inside bacteria , 16, 27.The Organization for Economic Cooperation and Development guides the health hazards of ZnO NPs/MPs based on their unique physicochemical properties obtained through synthesis procedures . The US The study goal is to investigate the cytotoxic effect of ZnOnps, made through a green synthesis, on ovarian cancer, as well as their antimicrobial activity against gram + ve and gram -ve bacteria, to provide a new treatment option for both cases.et al. [ZnOnps were obtained through the green synthesis method using green tomato extract Fig. , as prevet al. , 30. Culet al. , 31. The2 atmosphere at 37°C.SKOV3: Ovarian cancer was obtained from Nawah Scientific Inc., Mokatam, Cairo, Egypt. Cells were preserved in RPMI media supplemented with 100 mg/mL of streptomycin, 100 units/ml of penicillin, and 10% heat-inactivated fetal bovine serum in humidified, 5% (v/v) CO3 cells) were incubated in full medium for 24 h. Cells were then treated with a second portion of 100 μL medium containing drugs at varying concentrations. Cells were fixed by replacing the medium with 150 μL of 10% TCA and incubated at 4° C for 1 h after 72 h of drug exposure. After removing the TCA solution, the cells were washed five times with distilled water. Aliquots of 70 μL SRB solution (0.4%w/v) were added and incubated in a dark place at room temperature for 10 min. Plates were washed three times with 1% acetic acid and allowed to air-dry overnight. Then, 150 μL of TRIS (10 mM) was added to dissolve the protein-bound SRB stain; the absorbance was measured at 540 nm using a BMGLABTECH®- FLUOstar Omega microplate reader  [Sulforhodamine B (SRB) assay was used to evaluate the cell viability. In 96-well plates, aliquots of a 100 μL cell suspension (5 × 10Germany) .Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were obtained from the Microbiology Department, Faculty of Agriculture, Minia University.Pathogenic bacteria: Staphylococcus aureus and Escherichia coli as models for gram + ve and gram -ve bacteria, and zinc oxide (ZnO) was used as a control. The agar well diffusion technique method by Lorentz et al. was employed to investigate the antibacterial activity of the ZnOnps [The biological activity of the prepared ZnOnps was tested against e ZnOnps . Using aTo assess the performance of ZnOnps, our antibiotics novobiocin & cefadroxil  and neomycin & cefaclor  at a concentration of 30 μg were tested against the same strains by using disk the diffusion method .Every assessment was performed at least three times. Means and standard deviations were calculated. Data were analyzed by one-way ANOVA analysis followed by the Tukey test for multiple comparisons to determine the differences between groups using GraphPad Prism analysis software.The cytotoxicity of ZnOnps was investigated using SRB against the SKOV3 cell line that was cultured in 96 wells plates. Various concentrations of  μg/mL of the ZnOnps were used in the cell lines and tested after being incubated for 72 h at 37◦ C in 5% CO2. 50% inhibitory concentration (IC50) was noted at various ZnOnps concentrations. The cytotoxic activity of ZnOnps was determined by an SRB assay by calculating a cell viability percentage. The synthesized ZnOnps demonstrated concentration-dependent cytotoxic behaviour in ovarian cancer cells. Phyto-synthesized ZnOnps have caused IC50, at a concentration of 27.45 μg/ml, of anticancer activity to SKOV3-cells. Figure Viability% = OD sample/ OD control × 100.Staphylococcus aureus and Escherichia coli were selected as gram + ve and gram -ve bacteria models, respectively.In light of ZnOnps having a potential antibacterial action, S. aureus, the diameter of inhibition ranged between 20.16 ± 0.16 and 27 ± 0.57 mm. The inhibition zone diameter expanded with the gram -ve bacteria, in which the inhibition zone ranged from 25.66 ± 0.33 to 31 ± 0.33 mm. Furthermore, our findings revealed that biosynthesized ZnOnps showed a strong antibacterial impact on the microorganisms examined.Concentrations of 20, 40, 80 and 100 μg/ml were attributed to bacterial growth inhibition Figs.  and 4. TStaphylococcus aureus and Escherichia coli using the disk diffusion method  [P < 0.05). There was no significant variation in cell viability between 12 and 24 h after exposure to a lower concentration of 5 and 10 μg/mL. Overall, a significant effect was detected at greater doses of 20 and 30 μg/mL after 24 h; nevertheless, a substantial change in cell viability was observed when the concentration increased from 20 to 30 μg/mL. As a result, 20 and 30 g/mL are likely feasible doses for determining the impact of ZnOnps on ovarian cancer cells.Bai  < 0.05) . Althouget al. demonstrated the ZnOnps toxicity on human epidermal keratinocyte HaCaT cells after 24 h of exposure to concentrations of 0,10,20,40 and 80 μg/mL [et al. showed how human coronary artery endothelial cells responded to two distinct sizes of ZnOnps, such as 20 and 90 nm [Lee 80 μg/mL . The resnd 90 nm .et al. study [et al. [In the present study, it is realized that the ZnOnps with the size of 51.6 ± 3.6 nm have slight cytotoxicity at concentrations up to 8 μg/mL around 95.6 ± 2.04 to 98.8 ± 1.27% determined as viability percentage. However, at higher concentrations, significant toxicity has been observed. At 80 μg/ml, only 5.99 ± 1.60% of the cancer cells were still viable, and the IC50 was at a concentration of 27.45 μg/ml. These results are consistent with Gu l. study , where t [et al. . They stet al. discovered that the ZnOnps treatment reduced the antioxidative ability of ovarian cancer cells and increased proteotoxic stress [Production of ROS has previously been demonstrated to be a characteristic of ZnOnps –46. The Continuous breakthroughs in green nanotechnology, as well as the encouraging findings of this study, point to the use of rod-shaped nanostructures for therapeutic purposes . PreviouCucumis melo-assisted ZnOnps demonstrated stronger antibacterial activity against both types of bacteria (S. aureus and E. coli), which focuses on the substance that can penetrate the cell membrane. Among the two types of bacteria, E. coli has a thick impermeable cell wall lipopolysaccharide that is not easily destroyed by chemical compounds. However, the generated ZnOnps disrupt this cell membrane and inhibit the growth region, presumably due to the appearance of Zn2 + during surface reactivity or the creation of Reactive Oxygen Species and cell wall destruction. The size of the nanoparticles is also important in blocking the membrane of the bacteria. According to the findings, physical interaction between bacteria and metal nanoparticles initiated antibacterial activity [Depending on how the cell wall reacts with the nanoparticles, metal oxide nanoparticles generally have a strong antibacterial effect against any type of bacteria, whether gram + ve or gram -ve. However, activity . ZnOnps activity .E.coli when it was cultivated with 100, 250 and 500 μg/ml of α-Fe2O3@ZnO nanocomposite [et al. reported that the inhibition zone observed for E. coli was 6.70 ± 0.46, higher than those obtained for S. aureus, 6.20 ± 0.43 mm at 100 μg/ml [According to our results, multi-resistant S. aureus was less inhibited than omposite . In the 00 μg/ml . These rE.coli strain incorporated in this study which appears to have an inhibition zone of 21 ± 0.58 mm.Novobiocin’s mode of action is to inhibit DNA gyrase by binding the ATP-binding site in the ATPase subunit. Although novobiocin is generally effective against gram + ve bacteria, it is reported that it can be used for sensitive E.coli that has a leaky outer membrane, as liposaccharides in the outer membrane represent a barrier . This maet al. (2016) tested 68 S. aureus isolates and established that about 57.4% of S. aureus were sensitive to neomycin by using the disk diffusion technique. These results confirmed that the tested S. aureus was sensitive to neomycin but selectively to gram -ve bacteria [E.coli was estimated to be about 87.15% [Neomycin belongs to aminoglycosides, which inhibit bacterial protein synthesis. It effectively works gram -ve bacteria and is commonly used for perioperative prophylaxis . Bessa ebacteria . In 2012t 87.15% .E.coli isolates from different clinical specimens in children in 2009 were sensitive to cefaclor [S. aureus and E.coli attributed in our work were resistant to cefadroxil.Cefaclor, a second-generation cephalosporin, works similarly to penicillins. As a beta-lactam antibiotic, it inhibits the third and last stage of bacterial cell wall synthesis. It acts on both gram + ve and gram -ve bacteria . About 7cefaclor . Cefadrocefaclor . S. aureS. aureus and E.coli; 42.6% of S. aureus were resistant to neomycin. In 2012–2017, neomycin-resistant E.coli was estimated at about 12.85%, concerning cefaclor, 2,6.4% of E.coli isolates from different clinical specimens in children in 2009 were resistant [E.coli revealed that 31, 65, 60, 85, 0, 20, 33, 5, 6, and 47% of the strains were resistant to Ceftazidime, Cefuroxime, Ceftriaxone, Amoxicillin, Meropenem, Gentamicin, Ciprofloxacin, Cefoperazone-sulbactam, Amoxicillin-clavulanic acid and Sulphonamides, respectively [Many reports discussed antibiotic resistance of esistant , 56. A sectively .et al. (2022) studied 92 S. aureus and 34 E. coli isolates and reported that all the isolates were multi-drug resistant (MDR), resistant to equal or more than three antimicrobial classes. For S. aureus isolates, the maximum resistance was recorded for tetracycline (76.09%), oxacillin, and ampicillin (70.65%). More than 58% of the isolates exhibited resistance against erythromycin, amoxicillin-clavulanic acid and streptomycin. Regarding E. coli, the highest resistance was observed against oxacillin (64.71%) and ampicillin (58.82%) antibiotics. Oproxamitelly 30% of E. coli isolates were resistant to ceftriaxone, ciprofloxacin and cefaclor [Rana cefaclor . That suet al. (2022) nominated ZnO NPs to be used as an alternative antibacterial agent against β-lactam-resistant gram-ve pathogenic bacteria [Krishnamoorthy bacteria . These fHowever, eco-friendly technologies based on using fungi, algae, and bacteria to synthesize nanostructures are more popular than physical and chemical resources , as the in vitro drug release of hydrogels were studied at three different pH levels: 4, 6, and 8 [The systems were studied for these particles in drug delivery applications in terms of hydrogel rheology and polymer crosslinking degree. Farzaneh Sabbagh and colleagues employed acrylamide-based hydrogel drug delivery systems to release acyclovir from magnesium oxide nanocomposite hydrogels . Acyclov6, and 8 .ZnOnps synthesized from plants may be an essential research topic in the biomedical sector. The green synthesis of ZnOnps using green tomato extract has been highlighted in this study, as it is a simple, rapid, environmentally safe, and relatively low-cost process. This method can help to overcome the limitations of conventional chemical and physical methods.ZnOnps are considered a novel generation of antimicrobial agents; their wound healing and antibacterial characteristics stimulate their use after surgical procedures. In addition, their cytotoxic action on SKOV3 cells paves the way for a mutant treatment in gynaecological applications.Our recommendation is to investigate the toxicity of the green synthesized ZnOnps through in -vivo studies on experimental animals to determine the effect of these nanoparticles on normal cells and then decide whether it can be used systemically or only locally as identifying means to reduce unwanted toxicity.


P. aeruginosa and B. cereus. The content of PACs was used as an indicator for assessment of the influence of hydrophilic extracts on pancreatic lipase activity. Under normal physiological conditions, in the presence of bile, the extract, which contained 42.4% of PACs was more effective compared to the extract which contained 17.5% of PACs. At all concentrations (0.2–40 mg of sample/g of pancreatic lipase), it inhibited lipase activity by 33%. Purified PACs were the most effective in inhibiting lipase activity (by 36%). However, in pathological physiological conditions (without bile), the opposite effect on lipase activity was observed. Thus, PACs and extracts can be used as inhibitors of pancreatic lipase only under normal physiological conditions.The twigs of sea buckthorn, blackcurrant, gooseberries, quince, and grapes were evaluated as a promising source of biologically active compounds—proanthocyanidins (PACs). Sea buckthorn twigs had the highest content of PACs (9.2% on dry biomass). Preliminary pretreatment of biomass with freon R134a did not allow an increase in PACs content in the composition of hydrophilic extract but confirmed the value of freon extract as an antibacterial agent against  Moreover, the content of some of the secondary metabolites is even bigger in their lignocellulosic biomass than in fruits . As all than CO2 ,4, cultithan CO2 ; goosebethan CO2 ,7, grapethan CO2 , and quithan CO2 . The fruthan CO2 ,11, whicthan CO2 . PreviouThe target PAC properties studied in this research were connected with the anti-diabetic and anti-obesity properties of PACs, since obesity is one of the most influential diseases today. This is mainly caused by excessive consumption of sugar and high fat foods when all consumed energy is not used through physical activity and is stored by the body as fat. According to the World Health Organisation, around 2 billion adults currently are overweight, of whom 650 million are affected by obesity (BMI ≥ 30 kg/m²) ,14 with ®), lorcaserin (Belviq®), phentermine/topiramate (Qsymia®), naltrexone/bupropion (Contrave®) and liraglutide (Saxenda and Victoza®). All these drugs are not safe, as they have such side effects as oily discharge from the rectum, gas, imperative urge to defecate, steatorrhea, increased defecation, and fecal incontinence. In addition, increased blood pressure, high pulse rate, palpitation, dry mouth, headache, insomnia, memory impairment, and paraesthesia have been reported [A symptomatic solution to prevent obesity could be the action of digestive enzymes ,23. Pancreported .It was proven that PACs can inhibit digestive enzymes , with different inhibitory potency ,30. Sinc2, butane, and freons, has started to be developed extensively, mostly for plant material with a high content of lipophilic compounds  [2 and freon R134a were reported to be efficient for the extraction of cannabis, as the safest and highest-capacity processing methods [2 and R134a solvents can reduce the amount of supercritical fluid necessary [2 and is generally much cheaper. As an advantage of extraction by fluoro-hydrocarbons, the possibility of changing the yield of extractable compounds by the addition of a co-solvent is also mentioned [2 and freon are approved by the FDA as safe solvents and can replace hexane solvent, which is highly toxic and is banned for use in Europe. Using a closed loop in a freon extractor prevents the R134a global warming effect. The disadvantage of R134a freon extraction is the production stage of the freon from trichloroethylene. Therefore, supercritical CO2 extraction seems to be much “greener”. However, CO2 extraction demands much more electricity for heating, pressure, cooling, and intensive ventilation to prevent suffocation [2 capturing increases the fuel needs of an electricity plant by 25–40% [Sequential extraction allows the obtaining of both non-polar and polar compounds. Lately, extraction by liquified gases, such as supercritical COs, etc.) ,34. Gas s, etc.) . The liq methods . It was ecessary . Extractentioned . Both COfocation . CO2 capy 25–40%  which iny 25–40% . Thus, fBased on the analysis performed, the present study aimed to obtain the lipophilic and hydrophilic extracts and PACs from the twigs of five types of fruit trees/shrubs , widespread in the Baltic States and Europe, the chemical characterization of the obtained extracts, and the study of the effect of extracts and isolated PACs on the activity of the pancreatic lipase in normal and pathological patient conditions. The anti-microbial properties of lipophilic compounds obtained by hexane and freon were tested for preliminary validation of the concept of natural materials-based complex solutions for obesity patients. To the best of the authors’ knowledge, this is the first comparative study of the above-mentioned lignocellulosic biomass. Thus, the novelties of the present study are the usage of the above-mentioned lignocellulosic agro-waste as a source of extracts and PACs; evaluation of freon extraction as a method to obtain polar and semi-polar compounds from lignocellulosic biomass; study of the effect of co-solvent; anti-microbial properties of lipophilic extracts; and analysis of the influence of hydrophilic extracts and PACs on lipase activity.Hippopae rhamnoides L.), cultivar ‘Maria Bruvele’, Institute of Wood Chemistry (IWC) sample storage No. 91/03-20/146-22, were collected from the sea buckthorn plantation area in Seme parish, Tukums county of Latvia ; blackcurrant (Ribes nigrum L.), cultivar ‘Selechenskaja’, IWC No. 97/03-20/146-22; gooseberries (Grossulariaceae family), cultivar ‘Kuršu Dzintars’, IWC No. 98/03-20/146-22; quince (Chaenomeles japonica), cultivar ‘Rasa’, IWC No. 99/03-20/146-22; and grapes (Vitis vinifera), cultivar ‘Zilga’, IWC No. 100/03-20/146-22, were collected from the fruit-tree/shrub plantation area of Baldone parish, Kekava county of Latvia : 56.82065, 24.27653). Twigs were cut in September 2022. All biomass samples are deposited in the Institute of Wood Chemistry (IWC), room 239, at the freezer, and available for analyses. The twigs were dried at 20–25 °C temperature, leaves were removed, and the twigs were grounded with a mill . The grounded twigs biomass (further in the text—biomass) was stored at −8 °C.The twigs of sea buckthorn , ABTS+• , the reference antioxidants Trolox , as well as analytical standards gallic acid (purity ≥ 97.5%) and procyanidin B2 (purity ≥ 90%) were purchased from Sigma-Aldrich . Reference microbial strains: S. aureus MSCL 3340, P. aeruginosa MSCL 3314, E. coli MSCL 332, B. cereus MSCL 330 were received from the Microbial Strain Collection of Latvia .The solvents (high purity), DPPHThe scheme of extracts’ isolation from dry biomass (DB) is shown in Lipophilic extracts . The extracts, after ethanol evaporation, were freeze-dried to yield a DE. The yield of the DE is presented as a percentage based on ODB. The CI for the results did not exceed 3% at α = 0.05.Hydrophilic extracts  content in the extracts  concentrated HCl in n-butanol) and 0.2 mL of iron reagent (w/v) (FeNH4(SO4)2∙12 H2O in 2 N HCl) were added; the tube was vortexed and placed into water bath at 80 °C for 50 min. The absorbance of the mixture was then recorded at 550 nm with a UV/VIS spectrometer Lambda 650  against a blank containing 1 mL of extraction solvent. Each sample was analyzed in triplicate, and assay results were expressed as a percentage per DE. The CI for the results did not exceed 3% at α = 0.05).The content of PACs in extracts (WE 1-WE 5 and ET 1-ET 5) and purified PACs fraction were measured by the butanol-HCl method using procyanidin dimer B2 as a reference compound. An aliquot (1 mL) of the extract was transferred into the test tube, 6 mL of acid butanol (5% (2) solution (0.12 mL) was added and incubated for 5 min at room temperature, after which 10% aluminium nitrate solution (0.24 mL) was added to the mixture. After 6 min, 1 mol·L−1 sodium hydroxide (0.8 mL) was added. The absorbance was measured at 420 nm, and the result was expressed as mg of quercetin equivalents per g of dry weight (DW).Next, 5–10 mg of the DE was dissolved in 25 mL of 96% ethanol. Then 0.4 mL of the extract solutions were added to a 10 mL test tube containing water (2 mL). After that, 5% sodium nitrite . The second part of the extract was shaken with hide powder CRS and filtered. The phosphomolybdotungstic reagent was added to the filtrate, and after pH adjustment and 30 min of reaction time, absorbance was measured at 760 nm. As a standard substance, pyrogallol was used after a reaction with a phosphomolybdotungstic reagent. Polyphenols adsorbed by hide powder (=tannin content) were calculated by subtracting the unabsorbed polyphenols from the total polyphenols, each referring to the absorption of the standard substance. The tannin content was expressed as a percentage per DE. The CI for the results did not exceed 3% at α = 0.05).v/v; 2:8) with an approximate concentration of 0.1 mg·mL−1 after which it was filtered and then used for TOF/MS experiments. The TOF-MS spectra of PACs were recorded with Waters Acquity UPLC HClass with a PDA detector and Micromass QuattroMicro Mass spectrometer  using the Acquity UPLC BEH Amide column .Dry purified PAC fraction was dissolved in aqueous methanol (−1 and a split ratio of 1:30). Dry extracts  were dissolved in hexane (w/w 1:10) with an approximate concentration of 0.1 g/g hexane and filtered , after which they were used for GC-MS experiments. The oven program: 1 min isothermal at 60 °C, followed by 6 °C min−1 to 270 °C, and the final hold at 270 °C for 10 min. The mass spectrometer was operated in electron impact mode using 70 eV electron energy. The identification of the individual compounds was performed based on GC/MS using Library MS NIST 11 and NIST 11s, whereas the relative area of the peak of individual compounds was calculated using Shimadzu software based on GC/FID data. The summed molar areas of the relevant peaks were normalized to 100%, and the data for four repetitive pyrolysis experiments was averaged. The variation coefficient of measurement was ≤5%.Lipophilic extracts (HE 1 and FRE 1) were analyzed by GC-MS chromatography using Shimadzu GC/MS/FID-QP ULTRA 2010 apparatus , a capillary column RTX-1701  and a 60 m × 0.25 mm × 0.25 mm film  using a UV/VIS spectrometer Lambda 650 . The DPPH• assay was measured according to the procedures described by Dizhbite et al. [• (1·10–4 mol∙L−1) solution. DMSO was used as a control and Trolox as a reference antioxidant standard. CI ≤ 0.3 mg·L−1.The dry extracts  and purified PAC fraction were tested for their radical scavenging activity against the 2,2-diphenyl-1-picrylhydrazyl  using UV/VIS spectrometer Lambda 650 . ABTS+• was produced by the reaction of 2 mmol·L−1 ABTS stock solution with 70 mmol·L−1 potassium persulfate (K2S2O8), allowing the mixture to stand in the dark at room temperature for 12–16 h before use. The ABTS+ solution (stable for 2 days) was diluted with phosphate-buffered saline (pH 7.4) to an absorbance of 0.80 ± 0.02 at 734 nm. The absorbance at 734 nm was investigated 10 min after the mixing of 30 µL of extract (or purified PACs fraction and antioxidant standard—Trolox), diluted in DMSO of five different concentrations with 3.0 mL ABTS+• solution. DMSO was used as a control and Trolox as the antioxidant standard. CI ≤ 0.3 mg·L−1.The dry extracts (WE 1-WE 5 and ET 1-ET 5) and purified PAC fraction were tested for their radical scavenging activity against the 2,2′-azinobis  in 96-well microplates. The microbial suspension containing approximately 5 × 105 colony-forming units (CFU)/mL was prepared from a 24 h culture plate. From this suspension, 100 μL was inoculated into each well. The microtiter plates were incubated at 37 °C, for 24 h. After incubation, 40 μL of a 0.4 mg·mL−1 solution was added to each well as an indicator of microbial growth. The plates were incubated for 30 min and the MIC values were determined.Minimum inhibitory concentration (MIC) determination by microtiter broth dilution method was used. Stock solutions of the respective plant extracts were prepared in 10 mL microcentrifuge tubes by dissolving dry plant extract in dimethyl-sulfoxide (DMSO) to a final concentration of 100 mg·mL−1.Minimum bactericidal concentration (MBC) was recorded as the lowest concentration of an antimicrobial substance that reduces the viability of the initial microorganism inoculation by ≥99.9% after 24 h incubation at 37 °C. MBC was evaluated for extracts and purified PACs fraction. 10 μL were taken from the well obtained from the MIC experiment , as well as two wells above the MIC value well, and spread on Mueller–Hinton agar plates. The number of colonies was counted after 18–24 h of incubation at 37 °C. The concentration of sample that produces <10 colonies was considered as MBC value. CI ≤ 0.3 mg·L−1 (0.2–400 mg of sample/g of pancreatic lipase—further in the text—mg·g−1 PL) were added. All tubes were put in the incubator for 40 min at 38 °C. Afterwards, each solution from the tubes was titrated with 0.1 N NaOH solution until the colour changed into yellowish-brown, and the needed amount of NaOH was determined. CI ≤ 0.3 mg· g−1 PL.The test tubes were prepared composed of 4.0 mL milk (3.8% fat), 1 mL pancreatic solution , 1 mL bile , and 1 mL phenolphthalein. Orlistat , a known inhibitor of pancreatic lipase, was used as a reference compound. The first test tube was used as a control tube without the investigated components . To the following tubes, 100–2000 μL of the analyzed sample  at the concentrations of 2, 20, and 200 mg·LT distribution was calculated at a significance level of 5% (α = 0.05).All measurements were conducted in triplicate and the results were presented as the mean value. Statistical analyses were performed using Microsoft Excel 2016, version No. 16. CI for a mean using Student’s −1 DE).The yields of lipophilic extracts obtained with hexane from the entire studied biomass were quite close, but statistically different, and varied from 0.92 to 1.44% per DB. The yields of hydrophilic extracts from the same biomass, obtained using distilled water as the most environmentally friendly solvent, differed statistically significantly and varied from 7.1 to 15.7% per DB. With 50% EtOH, the extracts yield increased significantly, suggesting that the extractives are more soluble in the ethanol–water suspension. The yields of hydrophilic extracts obtained with 50% EtOH from the entire biomass under study were: sea buckthorn 21.7% > quince 14.3% > grapes 13.9% > black currant 13.5% > gooseberry 7.5% . The bio−1 DE. A comparative analysis of the solvent dependence on the yield of polyphenols and between biomasses is shown in Gooseberry biomass had the lowest yield of hydrophilic extracts (7.1% per DB by distilled water and 7.5% per DB by 50% EtOH). Polyphenols played the main role in the biological activity of all hydrophilic extracts. The amount of them in the 50% EtOH hydrophilic extracts was higher and varied from 30.3 to 48.6 g GAE·100 g−1 extract in 2020 [−1 DE in 2022). The literature data on TP of twigs of the other biomasses under study is very limited. Gooseberry berries were reported to contain TP in the amount of around 3 g GAE·100 g−1 biomass [−1 biomass for 50% EtOH extract). Quince berries were reported to be rich in PACs [The content of TP of sea buckthorn was very similar in years 2020 and 2022, considering the CI  in total flavonoid content (12.6 g RU·100 g−1 DW). All these polyphenols are biologically active natural antioxidants and antimicrobial agents, which can be used as ingredients in the formulation of different medications. The synergistic effect of the polyphenolic mixtures additionally leads to the simultaneous action on various disease pathways, which consequently contributes to a faster and more effective treatment outcome.Tannins are the dominant polyphenols in all hydrophilic extract compositions. The number of PACs, also known as condensed tannins, from the total amount of tannins ranged from 54 to 83% per DE. The highest content of PACs was observed for sea buckthorn 50% EtOH extracts (42.4% per DE). However, 50% EtOH extract from quince biomass was richer than sea buckthorn . The results were obtained in two fractions: a fraction consisting of low molecular polyphenols and their glycosides and free sugar, and a fraction consisting of PACs. The composition of PACs  was deteEvaluation of pre-purification of biomass from impurities was carried out using 1,1,1,2-tetrafluoroethane (Freon R134a). Based on the highest content of PACs in the extract , the seaSince this is the first research on freon extraction from twigs, there were study limitations consisting of seasonal and geographical changes in lipophilic compound content, which have to be further studied. The other types of freon  also have to be evaluated for the extraction. Although freon R134a is used in a close loop and has 0 ozone depletion, it has a high GHP (1300 vs. 7 vs. of the R1234ze) and its production and application could be banned after a while.Sea buckthorn biomass residue, after freon extraction with/without co-solvent, was extracted by 50% EtOH. The yield of hydrophilic extracts from all biomass residues slightly decreased. Small changes were seen only in the reduced content of polyphenols in 50% EtOH extract .LC-UV-ESI-QTOF-MS analysis (m/z 575), B-type of catechin/epicatechin trimer (m/z 865), gallo-catechin units (m/z 303), and catechin-epigallocatechin dimer (m/z 593)  .+ and DPPH• assays. In comparison to Trolox as a reference, which is a water-soluble derivative of vitamin E (IC50 = 4.0 mg L−1 in ABTS+• test and IC50 = 4.7 mg L−1 in DPPH• test), the 50% EtOH extracts from sea buckthorn and quince biomass showed the highest radical scavenging activity   .50 = 1.4 mg·L−1 in ABTS+• test and IC50 = 2.3 mg·L−1 in DPPH• test) than 50% EtOH extract from sea buckthorn (IC50 = 3.2 mg·L−1 in ABTS+• test and IC50 = 6.1 mg·L−1 in DPPH• test). The results for sea buckthorn twigs-derived PACs correlated well with the antioxidant activity of sea buckthorn twigs-derived PACs in year 2020: IC50 = 2.6 mg·L−1 in DPPH• test, with CI = 0.1 at α = 0.05 [−1 did not show the ability to inhibit DPPH and ABTS radicals.Due to the low content of polyphenols in blackcurrant, gooseberries, and grape extracts, the radical-scavenging activity was also low. Previous research has shown that purified PACs are powerful antioxidants. The radical scavenging activity of PAC fraction with PACs content 92.1% on dry weight (by Buthanol-HCl assay), was 2.3–2.7 times higher  showed significant inhibition of pancreatic lipase activity. Already at the amount of 0.2 mg of water extract containing 43.4 ± 0.4 g GAE·100 g−1 of polyphenols and 17.5 ± 0.1% of PACs, the lipase activity decreased by 22%. Further gradual increase of the amount of the water extract from 0.2 mg·g−1 PL to 40 mg·g−1 PL gave approximately the same lipase inhibition, within the confidence interval. The dependence of lipase activity inhibition on the content of PACs in the extract has also been established: the higher the content of PACs in the extract, the higher the percentage of inhibition. An extract containing 48.6 ± 0.2 g GAE·100 g−1 and 42.4 ± 0.3% PACs at the amount of 0.2 mg·g−1 PL to 40 mg·g−1 PL inhibited lipase activity by 33%. A PACs fraction (PACs content in extract 92.1% on dry weight) was the best pancreatic lipase inhibitor; at a concentration 1 mg·g−1 PL, pancreatic lipase activity decreased by 36%. At the same concentration, Orlistat was 1.4 times more effective (inhibited lipase activity by 52%). However, it has to be considered that Orlistat has side effects, described above in the Introduction. At the concentrations of PACs until 20 mg·g−1 PL, lipase inhibition was similar in inhibited value taking into account confidence interval. Therefore, in practical applications, the concentration 0.2–1 mg·g−1 PL is sufficient for lipase inhibition and there is no need to increase the concentration. When comparing all tested samples, the fraction of the extract after PAC separation (admixture) was not effective as a pancreatic lipase inhibitor, and at a concentration of 20 mg·g−1 PL contributed to the increase of lipase activity (by 6–11%) , all hydrophilic extracts of sea buckthorn at all concentrations (0.2–40 mg·gy 6–11%) . This co−1 PL; however, already at 2 mg·g−1 PL, pancreatic lipase activity activation was observed , a similar tendency was observed for lipase inhibition in the 50% EtOH extracts and purified PACs fraction at 0.2 and 1 mg·gobserved .−1 PL, PACs and extracts showed slight lipase inhibition activity. However, at 2 mg·g−1 PL almost all samples showed the neutral or lipase activation effects.In the PAC-free extract (admixture), the lipase activity under pathological conditions was higher (10–15%) compared to the results under normal conditions (6–11%). At the amount of samples’ of 0.2 and 1 mg·g−1 PL. The purified PAC sample at the amount of 2–400 mg·g−1 PL had the activation tendency of lipase activity in pathological conditions  and clinical studies, which are missing for almost all discovered lipase inhibitors.50 > 30 mg·L−1), but showed anti-microbial activity. The freon extract was more effective against P. aeruginosa and B. cereus, but the hexane extract was more effective against E. coli and S. aureus bacteria.Several fruit shrubs pruning plant material was evaluated and compared for the first time. It was shown that the lignocellulosic biomass originating from agro-waste of fruit shrubs is a source of valuable metabolites. Innovative freon extraction of non-polar and semi-polar compounds, as well as the influence of EtOH as co-solvent on further extraction of hydrophilic compounds, were tested. Lipophilic extracts obtained by freon had a different composition than those obtained by hexane and had a low antioxidant activity  in normal physiological conditions. In pathological conditions, both extracts, as well as PACs, tended to slightly increase lipase activity.Lignocellulosic biomass-derived PACs’ influence on pancreatic lipase proved in the study, along with PACs’ and the lipophilic extracts’ anti-microbial activity, can be used in complex treatments for obesity and its accompanying microbial infections. It has to be pointed out that due to inhibition of lipase, absorption of fat-soluble vitamins and nutrients is inhibited as well. Thus, lipase inhibition could be a temporary solution in fighting obesity, when patient’s pathological molecular mechanisms already developed and it’s not easy to change them just by transition to healthier lifestyle.