Patients with intraventricular tumors are more susceptible to postoperative meningitis (POM) than other intracranial tumors. In this study, we explored the risk factors of POM in lateral ventricular trigone meningiomas (LVTMs).Clinical features of 64 patients with LVTMs were analyzed. Age, gender, body mass index, medical history, intraoperative blood loss (IBL), intraventricular drainage placement, surgical duration, tumor grade, postoperative tumor cavity hemorrhage, and tumor size were included in univariate and multivariate analyses of POM.p = 0.003), tumor size ≥50 cm3 , and surgical duration ≥5 h  were considered possible risk factors for POM (p < 0.10). Tumor size (R = 0.514) and surgical duration (R = 0.624) were significantly correlated with IBL (p < 0.05). In the multivariate analysis, only IBL was found to be an independent risk factor for POM.Of the 64 patients, 14 patients (21.9%) received diagnosis of POM. The univariate analysis revealed IBL ≥400 mL (odds ratio [OR], 9.012; The IBL ≥400 mL is independently associated with the increased risk of POM in LVTM patients. Our results demonstrate the importance of controlling IBL for preventing POM, especially in large tumors and long surgeries. Meningioma is the second most common intracranial tumor in adults, with an incidence rate of 1.5–5.5/100,000 –9. Howevn = 64) to bring insight for reducing the incidence of POM.Postoperative meningitis (POM) is a challenging problem for all neurosurgeons due to severe complications. Moreover, the complications lengthen the hospital stay and increase the medical costs. Additionally, it can also cause serious neurological dysfunction and even death. Unfortunately, POM is difficult to treat. According to previous reports, the incidence of POM ranges from 0.8%–7% for all craniotomies and 19.8% for intraventricular tumor surgeries , 12. IntWe reviewed a total of 2,573 consecutive patients with the diagnosis of intracranial meningioma in the First Hospital of China Medical University from December 2010 to May 2018. Intraventricular meningiomas accounted for 2.8%  of all meningiomas.Exclusion Criteria: Patients with meningiomas in the third or fourth ventricle, lacking preoperative imaging data, and who had not received an operation in the First Hospital of China Medical University were excluded.Finally, 64 (2.5%) patients with LVTMs were included for the analysis. All of the patients underwent routine preoperative physical, radiological, and laboratory examinations. The study protocol was approved by the institutional review board of the First Hospital of China Medical University. In addition, written informed consent was obtained from each patient to use clinical data for future research.All patients received 1.0–2.0 g of intravenous ceftriaxone intraoperatively for infection prophylaxis as described previously –16. A se6/L), protein level (120–600 mg/L), and decreased glucose level (2.2–3.9 mmol/L) in the CSF; (3) organisms observed in the CSF smear examination; and (4) positive antigen test in the CSF.POM was determined as previously described . Briefly3 vs. ≥50 cm3) were included in univariate and multivariate analyses of POM.The clinical data were obtained from our hospital electronic information system. Patient information, including age (<50 vs. ≥50 years), gender, body mass index (BMI), past medical history (hypertension and diabetes), intraoperative blood loss (IBL), intraventricular drainage placement, surgical duration (<5 h vs. ≥5 h), tumor grade, postoperative tumor cavity hemorrhage, and tumor size . The chi-squared test was conducted, and factors with The study population was divided into two groups for validation analyses depending on the patient’s admission time. The first 32 patients from December 2010 to January 2015 were included in group A, while the last 32 patients, from March 2015 to May 2018, were considered group B.2, 50 cm3, 5 h, and 400 mL, respectively. There were 31 patients (48.4%) with age ≥50 years and 33 patients (51.6%) with age <50 years. The average tumor size was 52.78 ± 65.09 cm3 (1.9–301.4 cm3). There were 36 patients (56.2%) with tumors <50 cm3, while 28 patients (43.8%) with tumors ≥50 cm3. The surgery lasted for an average of 4.68 ± 1.88 h (2–11 h), whereas the average volume of IBL was 359 ± 342 mL . In 18 patients (28.1%) surgery lasted ≥5 h, while in 46 patients (71.9%) it lasted <5 h. There were 52 patients (81.2%) with IBL <400 mL and 12 patients (18.8%) with IBL ≥400 mL. There were 13 patients (20.3%) who had hypertension and no patients with diabetes. Intraventricular drainage was placed in 13 patients (20.3%). Eight patients had postoperative tumor cavity hemorrhage . Th. Th2, 503, and surgical duration ≥5 h were considered possible risk factors for POM . However, gender, age, BMI, and history of hypertension did not correlate with POM. Intraventricular drainage placement was not significantly associated with POM in this study.The potential risk factors of POM are presented in p = 0.018; P = 0.049, 0.046 for groups A and B, respectively).Furthermore, IBL, tumor size, and surgical duration were analyzed through multivariate analysis. As shown in p = 0.033).Patients with POM were treated with lumbar drainage and antibiotics . First, POM is a common postoperative complication of neurosurgery, causing severe consequences –25. PrevIn this study, 64 LVTM patients underwent craniotomy, out of which 14 patients (21.8%) had the diagnosis of POM. Previously, Wang et al. reported a 19.8% incidence of POM, 1.7% incidence of postoperative hematoma evacuation, and 3.3% incidence of postoperative ventriculoperitoneal shunt in out of 121 LVTMs –31. ThusWe found that IBL ≥400 mL was an independent risk factor for the POM. This observation was in-concurrence with previous studies reporting the correlation between IBL and POM , 34. TheIn addition, our results also indicate that large tumor size and long surgical duration were associated with increased IBL. This association further affects the incidence of POM. Large LVTMs are usually hypervascular and fibrous and require long-lasting operations. Moreover, cystic meningioma is a special kind of meningiomas with a relatively low occurrence of about 3.5% . There wOur results emphasized the importance of controlling intraoperative bleeding. Of note, preoperative embolization, meticulous intraoperative hemostasis, and early tumor devascularization may reduce the IBL and, in turn, decrease the risk of POM . HoweverThe current study had some limitations. One major limitation was the lack of distinction between bacterial and aseptic meningitis. As a result, they show similar symptoms , CSF test , except CSF culture and smear. However, CSF culture and smear show a low positive rate, which can be reduced by prophylactic antibiotics . In thisThe IBL ≥400 mL is independently associated with the increased risk of POM in LVTM patients. Our results demonstrate the importance of controlling IBL for preventing POM, especially in large tumors and long surgeries.


Blumea balsamifera (L.) DC., belonging to the Asteraceae family, also known as “ngai camphor,” is one of the traditional herbs used in Thailand for folk medicine and a component in local food and drinks. There was, however, no evidence indicating the presence of beneficial compounds at different leaf ages. Exploring various extraction solvents, we investigated the phenolics, flavonoids in particular quercetin content, antioxidant capacity, and antibacterial activity of immature and mature leaf extracts. The dried leaves were macerated in 50% ethanol, 95% ethanol, hexane, or decocted in water. Bioactive substances were analyzed by UV spectrophotometry and HPLC. Analysis of antioxidant capacity was done byDPPH, ABTS, FRAP, and NO scavenging assays. The antibacterial activity of immature leaf extract eluted with 50% ethanol was subsequentially evaluated in vitro. Extraction with 50% ethanol proved optimal, yielding 1.2–1.6-fold and 1.5-fold greater immature and mature leaf extracts than other solvents. More phenolics (1.2-fold), flavonoids (1.1-fold), quercetin content (4.8-fold), and antioxidant activity (1.3-fold) were found in the immature leaf extract. There was a significant positive correlation between antioxidant activity and bioactive compounds. The immature leaf extract eluted with 50% ethanol showed antibacterial activity against Staphylococcus aureus, with a minimum inhibitory concentration of 0.5 mg/mL. The immature leaves of B. balsamifera are a rich source of quercetin and phenolics, and 50% ethanol proved optimal for extracting bioactive components from these leaves. Herbs and plants native to the tropics have long been included in regional foods and beverages. In addition, their abundance of bioactive components, such as terpenoids, phenolics, and flavonoids, enables them to serve as therapies for a variety of human ailments and diseases. Plant phenolics consist of monophenols and polyphenols such as flavonoids, which function as antioxidants , 2. Of tBlumea balsamifera is an essential traditional Thai herb. In Thailand, the leaves have been used for relieving muscle spasms, appetizing in food, curing rheumatoid arthritis, postpartum medicine, expelling wind in the body, relieving colic, relieving stomachache, sweating, expectorant, reducing blood pressure, and driving parasites. Several studies on its phytochemistry have revealed its content of flavonoids, organic acids, esters, alcohols, sterols, and terpenes [B. balsamifera is a perennial shrub that usually develops new shoots in the rainy season. The young shoots can be made of tea drinks [B. balsamifera leaves.terpenes . After bterpenes . The driterpenes . The higterpenes . B. balsa drinks , whereasa drinks  and phota drinks . Becausea drinks . Immatura drinks . HoweverB. balsamifera leaves have been reported to contain highly bioactive compounds. For example, the essential oil obtained from Chinese B. balsamifera leaves contained major terpenoids of 1,8-cineole (21.0%), borneol (12.0%), β-caryophyllene (10.4%), camphor (8.1%), and 4-terpineol (6.5%) [B. balsamifera leaves for tea and topical applications. Traditional extraction methods include decoction and maceration by water and alcohol, often used in traditional medicine to obtain bioactive compounds from plants. Ethanol is generally recognized as safe for humans and is a suitable solvent for polyphenol extraction [Hypericum formosanum [Rosmarinus officinalis with high DPPH activity, ethanol at 50% was the most effective [B. balsamifera leaves were analyzed for some bioactive compounds and antioxidant capacity utilizing various extraction solvents. The antibacterial effectiveness of the optimum extract against particular human illnesses was then in vitro investigated.l (6.5%) , whereasl (6.5%) . Local ptraction . Neverthtraction . For exatraction , whereasrmosanum . When itffective . In the Ethanol, methanol, and acetonitrile were purchased from Daejung Co., Ltd. (South Korea). Hexane was provided by RCI Labscan (Ireland). 2,4,6-Tris (2-pyridyl)-s-triazine (TPTZ) was purchased from Fluka (Switzerland). Folin−Ciocalteu's phenol reagent, aluminum chloride hexahydrate, and sodium carbonate were brought from LOBA (India). Quercetin, Trolox , DPPH , ABTS  diammonium salt), potassium persulfate, gallic acid, and iron (III) chloride were supplied bySigma-Aldrich (U.S.A.). Trifluoroacetic acid was purchased from Fisher Chemistry (UK). Sodium hydroxide and aluminum chloride were purchased from Merck (Germany). All chemicals and reagents were of analytical grade.B. balsamifera containing tiny soft trichomes and soft surface on the dorsal epidermis (the 2nd–4th leaves from the shoot) and dark green mature leaves containing tiny stiff trichomes and matted surface on the dorsal epidermis  in 30 mL of deionized water for 15 min and by maceration in 30 mL of 50% ethanol (v/v), 95% ethanol (v/v), and hexane for 3 days at room temperature (25°C). After that, the solvents (ethanol and hexane) were evaporated using a rotary evaporator  at 40°C. The decoction was freeze-dried using the FD-1 freeze dryer . All extraction was done in triplicate. The crude extracts (10 mg) were dissolved in 1.5 mL of methanol and then analyzed for bioactive compounds and antioxidant capacity.Extracts of immature and mature μL) and 1.995 mL of the diluted Folin−Ciocalteu reagent were separately mixed in tubes. After incubating at room temperature for 4 min, 1.995 mL of 10% NaCO3 (w/v) was added. The mixtures were incubated in the dark at room temperature for 1 h. The blank was prepared by replacing each extract with methanol. Using a UV spectrophotometer , the absorbance was measured at 765 nm. The TPC was calculated from a calibration curve of gallic acid ranging from 100 to 2000 mg/L, with the equation y = 0.0003x + 0.0039 (R2 = 0.9992). The TPC in extracts was expressed as milligram gallic acid equivalent (GAE) per gram of dried sample (mg GAE/g DW).TPC in extracts was determined according to the Folin−Ciocalteu method . The extμL) and 0.2 mL of 5% NaNO2 were mixed and incubated at room temperature for 5 min. 0.2 mL of 10% AlCl3 was added, the mixture was stored for 6 min, and then 1 mL of 1 M NaOH was added. The solution was adjusted to 2 mL with deionized water and incubated for 30 min. The absorbance was measured at 510 nm using a UV spectrophotometer. Authentic quercetin was used for the standard curve at a range of 1,000–10,000 mg/L (y = 0.00008x + 0.0825 with R2 = 0.9996). TFC was expressed as milligram quercetin equivalents per gram dried sample (mg QE/g DW).TFC in extracts was measured according to Sultana et al.  with som18 5 μm, 4.6 ID × 250 mm., GL Sciences Inc., USA). The mobile phase consisted of deionized water (pH 2.5 adjusted by trifluoroacetic acid) and acetonitrile. The column was equilibrated with solvent A  and solvent B (acetonitrile) at a ratio of 85% and 15%, respectively. Solvent A was reduced to 15% for 50 min and then increased to 85% for 5 min. All ratios were operated within 60 min at a flow rate of 0.6 mL/min. Quercetin absorbance was detected at 320 nm. Quercetin solution was used to calibrate the standard curve at a concentration range of 0–400 mg/L (y = 40401x + 22911 (R2 = 0.9993). Quercetin content was expressed as microgram quercetin equivalents per gram dried sample (μg QE/g DW).Quercetin content of extracts was investigated using the method described by Crozier et al. . The extμL) was mixed with 3.990 mL of DPPH solution, and the mixture was shaken and kept in the dark for 30 min at room temperature. Methanol (10 μL) was used as the blank. The absorbance was measured at 517 nm using a UV spectrophotometer. Trolox solution was used for the standard curve at a concentration range of 40–2000 mg/L with the calibration equation y = 0.0004x + 0.0825 (R2 = 0.9998). DPPH values were expressed as milligram Trolox equivalents per gram of dried sample (mg Trolox/g DW).DPPH-free radical scavenging capacity of extracts was estimated according to Williams et al.  with sli+ solution was diluted with methanol until the absorbance of 0.700 at 743 nm. Then, 10 μL of the extract or methanol (blank) was mixed with 3.990 ml of the ABTS radical solution. The absorbance was spectrophotometrically measured at 734 nm. Trolox solution was used to calibrate the standard curve at 0–1000 mg/L using the calibration equation y = 0.0005x + 0.0026 (R2 = 0.9998). ABTS values were expressed as milligram Trolox equivalents per gram of dry weight (mg Trolox/g DW).Free radical scavenging activity of the extracts was determined by the ABTS radical cation decolorization assay . To prep3COONa buffer (pH 3.6), 20 mM FeCl3. 6H2O, and 10 mM TPTZ in 40 mM HCl at a ratio of 10 : 1 : 1. The extract (10 μL) was mixed with 3.990 mL of the FRAP reagent and incubated for 5 min at 37°C. For the blank, 10 μL of methanol was used instead. The absorbance was measured at 593 nm. Trolox solution was used to calibrate the standard curve at 0–1000 mg/L . FRAP values were expressed as milligram Trolox equivalents per gram of dry weight (mg Trolox/g DW).FRAP activity was determined following the modified method by Benzie and Strain . The FRAμL) was mixed with 680 μL of 2.6% sodium nitroprusside solution and 160 μL of PBS . The mixture was incubated at 25°C for 3 h in the dark, in which 20 μL of sulfanilamide solution (1% sulfanilamide in 5% phosphoric acid) was added, followed by incubation for 15 min. Next, 20 μL of NED solution (0.1% N-1-naphthyl ethylenediamine dihydrochloride) was added, and the mixture was kept in the dark for 15 min. The absorbance of the mixture was measured at 548 nm using a UV spectrophotometer. Nitrite was used to calibrate the standard curve at 200–10,000 μM (y = 0.00009x + 0.0325 with R2 = 0.9977). Nitric oxide inhibition values were expressed as μmol of nitrite equivalents per gram of dry weight (μmol NO2/g DW).Nitric oxide radical was generated from sodium nitroprusside and determined by the Griess reaction . The extin vitro for antibacterial activity. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were measured using the broth dilution procedure as described in modified CLSI M7-A7 [Staphylococcus aureus (ATCC 6538), the gram-negative Escherichia coli (ATCC 8739), and Pseudomonas aeruginosa (ATCC 9027) were used for this study. The antibacterial test was certified by the Expert Centre of Innovative Herbal Products (InnoHerb), Thailand Institute of Scientific and Technological Research (TISTR).In the present study, the immature leave extract obtained using 50% ethanol was analyzed SI M7-A7 . Three b8 CFU/mL. B. balsamifera leaf extract was diluted to the final concentration of 0.05, 0.1, and 0.5 mg/mLby MHB. Then, 50 μL of each inoculum was put in a tube containing 5 mL of the diluted extract and incubated at 35°C for 18 h for MIC detection. Cultures showing zero turbidity were chosen to streak on sterilized Mueller-Hinton agar (MHA) plates containing the different extract concentrations . All plates were incubated at 35°C for 18 h. The MBC was determined by observing plates for the absence of bacterial growth.The bacterial species were cultured in Mueller−Hinton broth (MHB) at 35 ± 2°C for 18–24 h. The bacterial culture was diluted with inoculum preparation media to achieve a suspension containing approximately to 1 × 10P < 0.05 and P < 0.01.All experiments were analyzed in triplicate. The obtained data were subjected to analysis of variance (ANOVA) using the general linear procedure of the Statistical Package for the Social Science (SPSS). The mean comparisons were made using Duncan's multiple range test (DMRT) at B. balsamifera leaves was significantly more than that which could be harvested from mature leaves. Out of all the different solvents that were tried, the one that produced the most immature and mature leaf extracts was ethanol with a concentration of 50%, followed by ethanol with a concentration of 95%, water, and hexane , a strong potential oxidant that can decompose to produce OH and NO2 [The antioxidant activity of r plants , 41–43. r plants . Moreover plants . The tox and NO2 . Free ra and NO2 . Plant e and NO2 .B. balsamifera leaves are used to treat bacterial infections. The immature leaf extract of B. balsamifera was evaluated for antibacterial activity against S. aureus, E. coli, and P. aeruginosa. Pathogenic bacteria, including our three species, cause several infectious diseases that threaten public health [E. coli can cause sepsis, diarrheal disease, and urinary tract infections [P. aeruginosa causes otitis media, keratitis, gastrointestinal infection, and pneumonia [S. aureus causes bacteremia, endocarditis, skin, and soft tissue infections [S. aureus was determined to be 0.5 mg/mL for the extract of immature leaves treated in 50% ethanol. In comparison, essential oil from mature Thai B. balsamifera leaves extracted with hexane showed a MIC of 1.2 mg/mL against S. aureus, according to research by Sakee et al. [As part of Thai traditional medicine, c health , 50. E. fections , P. aeruneumonia –54, and fections . In thise et al. . The cele et al. .B. balsamifera had higher TPC and TFC compared to the mature leaves, perhaps because of a biosynthetic rate and a lower breakdown process as found in Cistus ladanifer [Aronia melanocarpa and Vitis vinifera than in mature ones [Quercus semecarpifolia, Q. leucotrichophora, Q. serrata, Q. baloot, and mature leaves of Q. glauca, and all oak species had more tannins in the young leaves [B. balsamifera have a significantly greater quercetin content (6.9 mg/g DW), making them an excellent source of quercetin. The antioxidant, antibacterial, anti-inflammatory, and anticancer properties of quercetin have been demonstrated in vitro and in vivo [B. balsamifera could alternatively be not only for local medicine but a good component for making healthy food and drinks.Immature leaves of adanifer . Higher ure ones . TPC wasg leaves . In compg leaves , the you in vivo . Dietary in vivo . As contBlumea balsamifera is a good source of quercetin. Extraction with 50% (v/v) ethanol proved the most effective in terms of yield, bioactive compounds, and antioxidant capacity  of B. balsamifera leaf extract. The immature leaves contained higher phenolics, flavonoids, and quercetin contents than the mature ones. The immature leaf extract obtained with 50% ethanol showed antibacterial activity, with a minimum inhibitory concentration of 0.5 mg/mL against S. aureus. This information would aid in the cultivation of B. balsamifera plants for the use in traditional medicine and by pharmacists, as well as in the extraction process. The stimulation of increased production and accumulation of bioactive substances would be an additional area of study.


Microbial plankton represent a pivotal component of aquatic ecosystems worldwide. Despite their importance in pelagic ecosystems, microbial assemblages are less studied, in many Arctic regions. The study was concerned with the abundance of marine bacteria and viruses, in relation to a set of environmental variables, after the main productive season. We found a mosaic horizontal distribution of the microbial plankton, while the number of bacteria and viruses decreased with depth. Nutrients and zooplankton carbon were significant drivers of microbial abundance. Bacterioplankton abundance was positively correlated with counts of viruses, indicating close relations between these groups of microbes. Comparisons with previous studies revealed strong seasonal variations in the total abundance of marine bacteria, with the maximum values being in the summer and spring periods, while virioplankton counts were comparable in the autumn and spring–summer seasons. Our study is the first report regarding microbial plankton in the northeastern Barents Sea, in the autumn period, and provides baseline information, expanding current knowledge on the structure of pelagic Arctic ecosystems.5–10.7 × 105 cells·mL−1 and 0.86 × 106–14.98 × 106 particles·mL−1, respectively) and were correlated with each other. Bacterioplankton was dominated by small-sized cells , and the average volume of bacterial cells tended to increase toward the seafloor. The ratio of viral to bacterial abundance (VBR) was 11 ± 1 and did not differ between water masses and depth layers. VBR were higher, compared to summer values, suggesting a strong impact of viruses on bacterioplankton, after the main productive season. Redundancy and correlation analyses showed that inorganic nutrients (nitrate and phosphate) and organic carbon from zooplankton were most responsible for the total variability in the microbial parameters. Water temperature and salinity, also, had a measurable impact, but their influence was lower. Bacterial abundance was lower than in other seasons and regions of the Barents Sea, while viral abundance was comparable, suggesting a stronger viral impact on Arctic marine bacteria in the autumn season.In the marine environment, bacteria and viruses play a significant role in carbon fluxes, remineralization processes, and the infection of various organisms. We performed a survey in the northeastern Barents Sea, a region adjacent to the Arctic Ocean, to investigate spatial patterns of microbial plankton, after the main productive period, in October 2020. Two main water masses occurred in the study region—colder Arctic Water and warmer Barents Sea Water, representing transformed Atlantic Water. Multivariate analyses detected patchiness in the horizontal distribution of bacteria and viruses, and their abundances showed no clear association with the water masses. There was an obvious vertical pattern in microbial concentration, with the highest estimates in the upper layers. Surface viral and bacterial abundance varied in a wide range (2.20 × 10 Arctic ecosystems are very sensitive to changes in physical conditions and climatic variability; here, it is proposed to be more crucial, relative to lower latitudes ,2. ThereThe Barents Sea may be considered the most productive Arctic region ,2. The sThe pelagic ecosystem function and structure in the Barents Sea are, largely, determined by the hydrological regime, local circulation, frontal zones, seafloor topography, and general currents . Recent a concentrations, it can be higher than phytoplankton stocks . W. WDalnie studies ,29,30,31Samples for nutrient determination were collected into acid-cleaned polyethylene tubes, after thorough rinsing and filtration, through a 45 µm polycarbonate filter  inserted in a filter holder. Samples for nutrient analysis were collected in 20 mL vials and, immediately, frozen in liquid nitrogen and, then, stored at −20 °C to further analysis. Determination of dissolved-oxygen concentration was carried out using the Winkler method with an automatic-endpoint-detection burette Digital Burette VITLAB . Analyses of dissolved inorganic nutrients were carried out at the Murmansk Marine Biological Institute laboratory, according to standard methods .a determination with a volume of 5 L for each sampling layer were filtered, immediately, through 0.6 µm Vladiopore filters. The filters were kept frozen until the onboard analysis. The filters were, subsequently, extracted in 90% acetone, placed in a freezer at 4 °C for 24 h, and centrifuged, following the standard procedure [a concentrations were measured, using a Nicolett Evolution 500 spectrophotometer , which had been calibrated with commercially purified chlorophyll a preparations.Water samples for chlorophyll rocedure . Chlorop−3. Calculations of biomass of particular taxa were made using published mean individual wet, dry, or carbon weights as well as length/weight relationships [In the laboratory, the zooplankton organisms were identified and counted under an MBS-10 stereomicroscope. Abundance was expressed as individuals·mionships ,35,36. Aionships . The conionships .3), large cells (>2 µm), and chain cells (cell length-width ratio ≥10). The dimensions of at least 30 small cells in each sample were measured, and the average value  was calculated. All cells >2 µm were counted, directly, and their biovolume was calculated, according to shape. The wet bacterial biomass was converted to carbon biomass , using the following equation: C = 120 × V0.72, where V–average cell volume, µm3 [Water samples for marine bacteria were preserved with particle-free 40% formaldehyde  and, immediately, stored in slide boxes at −20 °C, for further microscopic analysis in the laboratory. Sub-samples (20 to 100 mL) were stained with DAPI for 8–10 min and filtered onto 0.2 µm Nuclepore filters using a low vacuum . The blaume, µm3 . Virus-l−4 cm; Ref. [3 cm2·d−1), and V and P are the viral and bacterial abundances, respectively. Additionally, we calculated the ratio of viral to bacterial abundance (VBR).Contact rates (R) between viruses and bacteria at each sample were determined from the equation: R = (Sh × 2π × w × Dv) × V × P , where Scm; Ref. ), D is ta concentrations, zooplankton abundance, and biomass) between different water masses and depth layers were tested using one-way ANOVA or the Kruskal–Wallis test and a post hoc Tukey test (p < 0.05). Assumptions of ANOVA were checked using a Kolmogorov–Smirnov test for normality and Levene’s test for homogeneity of variances. The Kruskal–Wallis test is the non-parametric analog of a one-way ANOVA, which does not make assumptions about normality. It is performed on the ranks of the measurement observations [p = 0.001) differences between water masses and depth layers [Differences in environmental parameters . To deth layers , with reh layers . Comparia content, zooplankton abundance, and biomass) and microbial data , constrained ordination procedures were performed in the CANOCO for Windows v4.5 software [To study the relationship between environmental variables . The aim of this approach is to summarize multivariate data, in a convenient way, in scatter diagrams . RDA is p-values, for multiple testing. In each case, a false discovery rate was set at 0.1. Means are presented with standard errors (±SE).The forward procedure of non-collinear environmental variables in CANOCO was used, with a significance level of 0.05. The statistical significance of each environmental variable was evaluated, by Monte Carlo permutation tests . RDA wasp < 0.05), except for the bottom layers (Water temperature varied in a wide range (−1.65 °C to 2.51 °C) and the mean values tended to decrease, from the southeastern to the northwestern part of the study area . The uppm layers . The sea−1), although there was a clear vertical trend, with the lowest values being recorded below 50–100 m (p < 0.001) in phosphate and oxygen concentrations, between water masses and depth layers, while nitrate concentrations were similar in both water masses but differed regarding sampling layers (p < 0.001). Phosphate and nitrate concentrations were significantly positively correlated with salinity and negatively correlated with water temperature . In contrast, oxygen concentrations tended to increase with temperature and decrease with salinity .Nutrient concentrations decreased from the southern to the northern part of the study area . This tr50–100 m . One-waya concentrations varied between 0.18 and 0.36 mg·m−3 and were, relatively, homogeneous in the study area (p < 0.1) were detected between temperature, salinity, nutrients, and chlorophyll a concentrations.Surface chlorophyll udy area . No clea−3 (st. 10) and 1278 ind.·m−3 (st. 13), averaging 1012 ± 87 ind.·m−3  , in the total zooplankton biomass  amounted to >50% of the total biomass in ArW .Zooplankton density ranged between 752 ind.·mind.·m−3 . The tot mgC·m−3 . Both pa< 0.001) . Copepod< 0.001) . The zoo biomass , while ls in ArW . The tot5 to 10.68 × 105 cells·mL−1 (5 cells·mL−1. The surface and bottom counts increased from the Novaya Zemlya coast to Franz Joseph Land (p < 0.001). The abundance showed two peaks at the intermediate layer (50 m) as well as stations 10 and 14 (−3 to 12.48 mgC·m−3 (mean 5.90 ± 0.33 mgC·m−3) and demonstrated mosaic distribution within the study area, although maximum records were found at stations affected by ArW   .6 particles·mL−1 to 14.98 × 106 particles·mL−1, averaging 6.10 ± 0.45 × 106 particles·mL−1 (p > 0.05), but the surface and intermediate layers had greater values . The viral abundance was positively correlated with the bacterial abundance and biomass . VBR was estimated to be 1–28 (11 ± 1) (p > 0.05). Contact rates (R) between viruses and bacteria were 0.3–9.6 (3.5 ± 0.3) (p < 0.001). Pearson correlation analysis found significant positive correlations of R, with viral and bacterial abundances , while R was negatively correlated with ABV .Viral abundance ranged from 0.86 × 10les·mL−1 c. There (11 ± 1) . This va5 ± 0.3)  and signp = 0.013), although there were no significant differences in the microbial density averaged for water masses . Based on the SIMPER analysis, dissimilarities ranged from 19.9% to 44.4%. The highest dissimilarity occurred for the intermediate and bottom layers. The SIMPER analysis, also, indicated that viral abundance contributed, mostly (92–93%), to the total dissimilarities between sampling layers.Multivariate analyses , based on the abundance of microbial plankton, revealed a high degree of similarity between the two types of water masses in the region . The ANOPossible relationships, between the microbial plankton and the environmental dataset, were analyzed using RDA analysis and supplemented with the Pearson correlation analysis . The firThe second axis was significantly positively correlated with zooplankton biomass and oxygen concentration and negatively correlated with salinity . The forThe present study aimed to fill the gap in our knowledge of microbial plankton, in a less studied region strongly affected by cold waters from the Arctic Ocean, after the main productive season. Our data is the first report describing spatial patterns in bacterioplankton and virioplankton, in the northeastern Barents Sea. In general, we revealed that microbial abundance was strongly affected by a set of environmental variables .A clear warming trend has been detected in the Barents Sea, since the 2000s, and the mean water temperature was found to have risen ,2,3,4. INutrient concentrations demonstrate strong seasonal and spatial fluctuations in the Barents Sea ,2,52. Toa concentrations are low and do not exceed 0.1–0.5 mg·m−3 [a values, which confirmed this general pattern, for phytoplankton density to decrease, after the main productive season in the northern Barents Sea.In our study, we recorded a clear vertical pattern in the nutrient concentrations, with the richest waters being located in deeper layers. This pattern can be explained by intense vertical mixing processes, in the upper 50 m layer, and convection, as revealed in earlier studies ,2. Enhan5 mg·m−3 ,54,56. WCalanus spp., Pseudocalanus spp., Microcalanus pygmaeus, Oithona similis, and Metridia longa being common taxa [a. This result is expected because most species in our study were found to be herbivorous or omnivorous, and phytoplankton, apparently, played an important role in their feeding in the Barents Sea and other Arctic regions [Our study found that zooplankton assemblages were dominated by copepods. The zooplankton composition was typical for ArW and BSW with mon taxa ,28,30,57mon taxa ,30,31,58mon taxa . Local vmon taxa ,30. We,  regions .In the high Arctic, prolonged periods of low water temperature, ice coverage, and darkness in the autumn–winter season lead to strong seasonal differences in bacterial abundance and biomass as well as concentrations of viruses ,18,19,606–4.1 × 106 cells·mL−1, during summer in the central Barents Sea, and these counts were lower than the estimates reported for the northern Barents Sea, during the spring bloom [6 cells·mL−1/66 mg·m−3 (ArW) and 0.6 × 106 cells·mL−1/41 mg·m−3 (AW), in mid-November 2013 [Many studies have reported marked spatial and seasonal variations, in microbial characteristics in the Barents Sea. Spring and summer patterns of bacterioplankton have been investigated in the Marginal Ice Zone and in the northern Barents Sea ,23,24,25ng bloom  and summng bloom . The autber 2013 . Our stuber 2013 . In the ber 2013 ,62, in cber 2013 . Spatialber 2013 . Other sber 2013 ,22,23. T6 and 8.6 × 106 particles·mL−1 [Virioplankton abundance in the Barents Sea, also, demonstrated seasonal fluctuations, with the highest values occurring in the summer period. In the central Barents Sea, the number of virus-like particles ranged between 0.66 × 10les·mL−1 . Similarles·mL−1 ,25. Autules·mL−1  were simles·mL−1 ,25. Our les·mL−1  was muchles·mL−1 ,65.We detected a positive significant correlation between the abundance of viruses and bacterial stock, and this pattern appears to be common in the Arctic marine environment . The varResults of the present study showed patchiness in the horizontal distribution of bacteria and viruses in the Barents Sea. This is a well-documented phenomenon, reflecting the discontinuous distribution of individual organisms in aquatic environments . Aggregaa, and zooplankton stock) accounted for 61% of the total fluctuation in microbial plankton. Three parameters were found to be significantly associated with bacterioplankton and virioplankton. The most important factor affecting the distribution of microbial plankton was nitrate concentration. Negative correlations between nutrient supply and microbial abundance are considered to be, indirectly, connected with phytoplankton. Chlorophyll a was found to be one of the main drivers, controlling both abundance and biomass of marine microbes in the Arctic, during the spring and summer seasons [a values were low, suggesting that phytoplankton biomass was lower than in the most productive season. Nevertheless, microalgae seem to be important organisms, utilizing inorganic nutrients in the upper layer. On the other hand, there were great concentrations of nitrate and phosphate in the bottom layer, where bacterial abundance was significantly lower. This, also, might explain negative correlations between inorganic nutrients and microbial plankton. The enhanced supply of organic matter increases bacterial growth and leads to an increase in viral abundance in the marine environment [The RDA revealed that a set of environmental variables . A clear vertical pattern in microbial abundance was evident, with the highest concentrations in the upper layers. Small-sized cells were the most dominant group of the bacterioplankton, and average bacterial-cell volume tended to increase, near the seafloor. Bacterial abundance was lower than in other seasons and regions of the Barents Sea, while viral abundance was comparable. Bacterioplankton abundance was positively correlated with concentrations of viruses, indicating close relationships between these groups of microbes. Nutrients and organic carbon of zooplankton, together with hydrological variables, explained most of the spatial variability in microbial parameters.Our study expands current knowledge on the structure of pelagic ecosystems in the Arctic and provides baseline information on marine microbes in the autumn season. This investigation may be used in future research, focusing on the ecology of Arctic plankton assemblages, during the period of global environmental changes.


The data of our study detail the accumulation of the phytochemical composition of biologically active compounds in cranberry samples during the stages of maturity, therefore these data are significant for the assessment of harvest time of cranberry and can be applied to select cranberry cultivars for further cultivation in Lithuanian climatic conditions.In our investigation, we evaluated the content of chlorogenic acid, proanthocyanidins, anthocyanins, flavonols, triterpenoids, and phytosterols in cranberry fruit extracts of the cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ grown in Lithuania, as well as changes in the antioxidant activity in extracts of fruit samples of these cultivars during the period of berry maturation. The highest amount of proanthocyanidins (8.87 ± 0.57 mg EE/g EE) and flavonols (3688.52 ± 22.85 µg/g) was determined in cranberries of the cultivar ‘Howes’ harvested on 12 August. Remarkably, the highest anthocyanins content (9628.62 ± 266 µg/g) was determined in cranberries of the cultivar ‘Howes’ harvested on 22 October. The study showed that the content of phytochemical compounds in cranberries varied between 12 August and 22 October; the content of proanthocyanidins decreased by a factor of about 2, the content of chlorogenic acid decreased by a factor of about 1.3, the content of flavonols decreased by a factor of about 2, and the content of anthocyanins increased by 27 to 450 times. A strong correlation was found between the total proanthocyanidin content of cranberry fruit extracts and their in vitro antiradical and reducing activity (r = 0.781 and 0.726, respectively,  Vaccinium macrocarpon Aiton) belongs to the Ericaceae botanical family [Vaccinium oxycoccus L.), which has resulted in the increasing popularity of the cultivation of large cranberries in the Baltic region [Large cranberry  determining the pharmacological effects in cranberry fruit samples. To this end, research has been carried out and innovative instrumental analytical methodologies have been developed, optimized, and validated to analyze the qualitative and quantitative composition of cranberry fruit samples. Vilkickytė et al. proposed and validated a UPLC-DAD methodology for the analysis of anthocyanins and anthocyanidins in cranberry and other fruit samples . SedbareThe aim of our study was to investigate the phytochemical composition of anthocyanins, flavonols, chlorogenic acid, proanthocyanidins, triterpenoids, and phytosterols in fruit samples of large cranberry cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ grown in Lithuania, in the collection of Vilnius University Botanical Garden, and to evaluate amount changes in these compounds during the ripening period of the fruit. Anthocyanins have been detected in cranberry fruit samples, and have antioxidant ,22 and aAnalysis of anthocyanins and anthocyanidins composition showed that cyanidin-3-O-galactoside, cyanidin-3-O-arabinoside, peonidin-3-O-galactoside, and peonidin-3-O-arabinoside accounted for 93.11% ± 3.4% of the sum of anthocyanins and anthocyanidins in cranberries. The composition of these four predominant glycosidic compounds of the anthocyanin group varied between August 12 and October 22: 36.89% ± 14.66% of cyanidin-3-O-galactoside, 25.84% ± 6.31% of cyanidin-3-O-arabinoside, 21.70% ± 10.78% of peonidin-3-O-galactoside, and 8.68% ± 4.46% of peonidin-3-O-arabinoside . The quaViškelis et al. studied cranberry samples of the cultivars ‘Pilgrim’, ‘Stevens’, ‘Black Veil’, and ‘Ben Lear’ grown in the Lithuanian climate and determined that the content of the major anthocyanins was 33.13% ± 1.15% of peonidin-3-galactoside, 6.21% ± 1.07% of peonidin-3-arabinoside, 21.28% ± 1.05% of cyanidin-3-galactoside, and 17.57% ± 1.05% of cyanidin-3-arabinoside [Our study showed that the lowest total amounts of anthocyanin (10.45 ± 0.7 µg/g–296.49 ± 10 µg/g) were determined in cranberry fruit samples collected on 12 August. The lowest amount of total anthocyanins (10.45 ± 0.7 µg/g) was determined in the cranberry cultivar ‘Baifay’ collected on 12 August and consisted of cyanidin-3-O-galactoside. The sum of anthocyanins in cranberries of the cultivar ‘Early Black’ harvested on 12 August was 28 times higher than that determined in cranberry samples of the ‘Baifay’ cultivar harvested on the same day. In cranberries of the cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ collected on 12 August, the levels of malvidin-3-O-galactoside, malvidin-3-O-arabinoside, and malvidin were below the limit of detection, with a more marked increase in the levels of these compounds starting on 10 September .The highest total amounts of anthocyanin were determined in samples of the tested cranberry cultivars collected on 22 October. The sum of anthocyanins in fruit samples collected on 22 October ranged from 3710.58 ± 89 µg/g to 9628.62 ± 266 µg/g. The lowest amount of total anthocyanins (3710.58 ± 89 µg/g) was determined in samples of the cranberry cultivar ‘Stevens’ collected on 22 October. The highest amount of total anthocyanins (9628.62 ± 266 µg/g) was determined in cranberries of the cultivar ‘Howes’. Cranberries of the cultivar ‘Howes’ harvested on 22 October contained significantly higher levels of delphinidin-3-galactoside, malvidin-3-O-galactoside, malvidin-3-O-arabinoside, malvidin, peonidin, and cyanidin compared to the levels determined in cranberries of the other tested cranberry cultivars harvested on 22 October . The content of anthocyanins and anthocyanidins in cranberries of the cultivars ‘Baifay’, ‘Early Black’, ‘Pilgrim’, ‘Howes’, ‘Red Star’, and ‘Stevens’ grown in Lithuania during the ripening period between 12 August and 22 October increased by 27 to 450 times . During The results of our study show that the cranberry cultivars ‘Howes’, ‘Early Black’, and ‘Pilgrim’ have the highest level of anthocyanins, so these cultivars are promising for growing in Lithuanian climates as a source of anthocyanins. Furthermore, the study revealed an increasing trend in anthocyanin and anthocyanidin content, which provides knowledge on the quantitative changes in individual compounds during the ripening period and allows for selecting the optimum time for cranberry picking when the berries accumulate the highest levels of anthocyanins. Studies on the chemical composition of cranberry fruit samples allow the qualitative and quantitative composition of individual anthocyanins to be assessed and allow for the preparation of useful cranberry raw material with the highest content of anthocyanins, the biologically active compounds that determine the biological effects of cranberries.Flavonols are biologically active compounds with antioxidant, cardioprotective, antibacterial, antiviral, and anticancer activities ,30,31. OWang et. el. determined that quercetin-3-galactoside contains 31–46%, myricetin-3-galactoside contains 19–32%, quercetin-3-α-L-arabinofuranoside contains 7–17%, and quercetin-3-rhamnoside contains 7–14% of the total identified flavonols content in cranberries of different cultivars . These rThe evaluation of flavonols showed that the total flavonol levels in cranberries ranged from 1465.56 ± 31.22 µg/g to 3688.52 ± 22.85 µg/g. The highest total flavonol content (3688.52 ± 22.85 µg/g) was determined in cranberries of the cultivar ‘Howes’ harvested on 12 August, and the lowest total flavonol content (1465.56 ± 31.22 µg/g) was determined in cranberries of the cultivar ‘Howes’ harvested on 22 October.During the ripening period of the fruits from 12 August to 22 October, the sum of the flavonols in cranberries decreased by 1.46 to 2.52 times . VvedensThe flavonol content in cranberries of the cultivar ‘Howes’ decreased throughout the berry picking period. Cranberry cultivars ‘Baifay’, ‘Early Black’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ showed a different trend in total flavonol content from that seen in the ‘Howes’ cultivar. In cranberry samples of cultivars ‘Baifay’, ‘Early Black’, ‘Pilgrim’, and ‘Red Star’, an increase in total flavonols by about 1.2 times was observed on 24 September . In fruiThe trends in the changes of the quantitative composition of the compounds myricetin-3-galctoside, quercetine-3-galactoside, and quercetine-3-arabinofuranoside found in the tested cranberry samples were similar to those of the total flavonol content. Between 12 August and 22 October, the amounts of myricetin and quercetin decreased by a factor of 3 and 4, respectively. Quercetin-3-rhamnoside content in fruit samples showed little variation, with a decrease of about 1.1-fold in cranberry samples of cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Red Star’, and ‘Stevens’, and an increase of about 1.15-fold in cranberry samples of the cultivar ‘Pilgrim’. Wang et al. determined that quercetin-3-galactoside and myricetin-3-galactoside levels varied only slightly during the different stages of maturity in cranberries of the cultivars ‘Howes’, ‘Ben Lear’, ‘Stevens’, Early Black’, ‘Crimson Queen’, ‘Mullica Queen’, ‘Demoranville’, and ‘#35′ .Studies on the phytochemical composition of cranberries allow the determination of flavonols, their quantitative composition, and trends in their content. The highest flavonol levels were found in fruit samples of the cultivars ‘Howes’, ‘Pilgrim’, and ‘Red Star’ taken at different ripening periods, indicating the importance of harvest time. The determination of the content of flavonols is important for assessing the quality of cranberry raw material and for predicting the biological effects of cranberry raw material preparations, which are determined by flavonols.The phenolic carboxylic acids found in cranberry fruit plant material are biologically active compounds. Oszmianski et al. suggested that concentration of phenolic acids, especially of chlorogenic acid is important, as these compounds are the precursors of flavour in cranberry fruit . SantanaIn our study, the analysis of chlorogenic acid content in cranberries showed that chlorogenic acid content varied from 104.73 ± 1 µg/g to 489.70 ± 3 µg/g A. The hiDuring the ripening period from 12 August 2 to 27 August, the chlorogenic acid content in cranberries of cultivars ‘Baifay’, ‘Howes’, ‘Pilgrim’, ‘Early Black’, ‘Red Star’, and ‘Stevens’ decreased, on average, by a factor of 1.33 . During The quantity of chlorogenic acid in cranberries shows little variation during the ripening period of the cranberry fruit, thus the qualitative and quantitative evaluation of chlorogenic acid content can serve as one of the markers for the identification of the cranberry raw material and the assessment of its quality.Escherichia coli [Helicobacter pylori infection [Cranberry fruit raw material was found to contain proanthocyanidin compounds, which are responsible for the antibacterial effect of cranberry preparations. Pérez-López et al. determined that a group of proanthocyanidins inhibit P-fimbriae synthesis and induce a bacterial deformation, on both antibiotic-susceptible and antibiotic-resistant uropathogenic hia coli . Li et anfection . In our During the cranberry ripening period from 12 August to 8 October, a decreasing trend of proanthocyanidin content was observed in cranberry samples of cultivars ‘Baifay’, ‘Howes’, ‘Pilgrim’, ‘Early Black’, ‘Red Star’, and ‘Stevens’, which showed a decrease of about a factor of 2. During the ripening period from 8 October to 22 October, the decrease in proanthocyanidin content in cranberry samples of the tested cultivars either stabilized or there was a 1.1-fold increase. Wang et al. and Vvedenskaya et al. studies confirm our results that the amount of proanthocyanidins decreased most intensively at the beginning of maturity, slightly increased at later stages of maturity ,27. SimiStudies on the amount of proanthocyanidins in cranberry maturity stages are relevant for the preparation of cranberry plant material. The determination of proanthocyanidin content provides an opportunity to prepare cranberries with a known proanthocyanidin content, which can be used for the production of cranberry preparations. Proanthocyanidins in cranberry fruit raw material provide antibacterial effects in the treatment and prevention of urinary tract infections, and the identification of these compounds is therefore important to produce high-quality food supplements and medicinal products of known composition and efficacy ,40.Triterpenoids identified in cranberry fruit are responsible for their anti-inflammatory, antitumor, and anticancer effects . Chen etIn our study, we identified and quantified triterpenoids  and phytosterols (campesterol and β-sitosterol). In cranberry fruit samples collected between August 12 and October 22, ursolic acid accounted for 75.62% ± 2.54%, oleanolic acid—for 19.50% ± 1.42%, corosolic acid—for 2.71% ± 1.95%, maslinic acid—for 0.89% ± 0.78%, α-amyrin—for 0.86% ± 0.45%, and β-amyrin—for 0.42% ± 0.32% of the total triterpenoid content. Klavins et al. confirmed that ursolic acid is the predominant triterpenoid in cranberry fruit [In cranberry fruit samples, β-sitosterol content was higher (98.16% ± 1.84%) than that of campesterol (1.84% ± 0.45%). Wu et al. also indicated that β-sitosterol is the major phytosterol in cranberry fruit . ŠedbarėThe changes in the total amount of triterpene compounds detected in fruit samples of the tested cranberry cultivars between 12 August and 22 October showed a mixed pattern . This vaOszmianski et al. found that the concentration of triterpenoids  in fruits of cranberry cultivars ‘Stevens’ and ‘Pilgrim’ collected from 1 September to 22 September increased by a factor of 1.3 and 1.1, respectively . In our The changes in the amounts of the detected triterpenoids  in cranberries varied. During the ripening period, the quantity of maslinic acid in cranberry fruits of ‘Early Black’ and ‘Howes’ cultivars was significantly higher than that in fruits of ‘Baifay’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ cultivars . During The quantity of corosolic acid in cranberries of cultivars ‘Early Black’ and ‘Howes’ was higher than that found in cranberries of cultivars ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ cultivars during ripening . BetweenThe quantity of α-amyrin in fruits of the tested cranberry cultivars increased by up to 40-fold between 12 August and 22 October . The conAnalysis of triterpenoids in cranberry plant material showed that the triterpenoid content in cranberry fruits varied during the ripening period. The high triterpenoids levels during the ripening of cranberry fruits were found in the samples of the ‘Early Black’ cultivar. This is possibly because the cranberries of this cultivar are smaller in size and have a larger surface area of the peel, where triterpenoids accumulate . TriterpThe determination of the phytochemical compounds is important in the preparation of cranberry plant material, as it allows for the preparation of material with known levels of triterpene compounds and phytosterols, which may influence the biological effects of the preparations.Phenolic compounds and triterpenoids found in plant matrices are natural antioxidants ,51. AntiIn our study, the antiradical activity of 100% acetone and 70% ethanol acidified with 1% HCl extracts of cranberries was estimated by the ABTS method, and the reducing activity was tested by applying the FRAP method. The extraction of triterpenoids by aqueous mixtures of organic solvents  is less efficient with a lower polarity organic solvent, and therefore 100% acetone was chosen as the extraction solvent to improve the efficiency of the extraction of triterpenoid compounds from cranberry fruit samples ,56. PhenChanges in the antioxidant activity of cranberry extracts occurring during berry ripening are shown in The antiradical activity of cranberries acetone extracts evaluated by the ABTS method during the ripening of the berries ranged from 3.39 ± 0.38 µmol TE/g to 14.90 ± 2.10 µmol TE/g. The reducing activity of cranberries acetone extracts estimated by the FRAP method ranged from 4.51 ± 0.26 µmol TE/g to 17.61 ± 3.26 µmol TE/g. The acetone extracts of cranberry cultivar ‘Howes’ produced from cranberry samples collected on 12 August showed the strongest antiradical (14.90 ± 2.10 µmol TE/g) and reducing (17.61 ± 3.26 µmol TE/g) activity.Acetone extracts of cranberry cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Pilgrim’, and ‘Red Star’ made from fruit samples collected on 12 August showed by 2 times stronger antiradical activity and by 1.5 times stronger reducing activity than acetone extracts of cranberry fruit collected on 22 October did. Acetone extracts of fruit samples of the ‘Stevens’ cultivar showed a 1.5-fold decrease in antioxidant activity between 12 August and 24 September, and an approximately 1.5-fold increase between 24 September and 22 October.The antiradical activity of the ethanolic cranberry extracts varied from 95.56 ± 3.74 µmol TE/g to 223.90 ± 16.11 µmol TE/g during the ripening of the berries. The extracts of the ‘Howes’ cranberry fruit samples collected on 12 August demonstrated the strongest antiradical activity (223.90 ± 16.11 µmol TE/g). The weakest antiradical activity (95.56 ± 3.74 µmol TE/g) was determined in cranberry extracts of the ‘Stevens’ cultivar made from cranberry samples collected on 22 October.The antiradical activity of the ethanolic cranberry extracts of the cultivars ‘Early Black’ and ‘Howes’ decreased by a factor of about 1.4 between 12 August and 8 October and increased by a factor of 1.3 between 8 October and 22 October. The strongest antiradical activity of the ethanolic cranberry extracts of cultivars ‘Pilgrim’ and ‘Stevens’ was found in extracts made from cranberries harvested on 10 September. The ethanolic extracts of the cranberry cultivars ‘Baifay’ and ‘Red Star’ showed only a slight variation in antiradical activity during the study.The reducing activity of cranberry ethanolic extracts as assessed by the FRAP method varied from 100.79 ± 13.46 µmol TE/g to 223.60 ± 14.96 µmol TE/g. The strongest reducing activity (223.60 ± 14.96 µmol TE/g) was observed for the ethanolic extracts of cranberry fruit samples of the cultivar ‘Howes’ collected on 27 August. Meanwhile, the ethanolic cranberry extracts of the cultivar ‘Pilgrim’ harvested on 12 August had the weakest antiradical activity (100.79 ± 13.46 µmol TE/g).The reducing activity of the ethanolic cranberry extracts of the cultivars ‘Early Black’, ‘Howes’, and ‘Stevens’ decreased by a factor of about 1.3 between 12 August and 8 October and increased by a factor of about 1.2 between 8 October and 22 October. The strongest reducing activity of the ethanolic extracts of the cranberry cultivars ‘Pilgrim’ and ‘Red Star’ was found in the extracts from cranberries harvested on 27 August. The antiradical activity of the ethanolic fruit extracts of the cranberry cultivar ‘Baifay’ showed only a slight variation during the study.p < 0.001 and r = 0.726, p < 0.001, respectively). Proanthocyanidins identified in cranberry fruit samples contain epicatechin units in their structure, which have exceptionally strong antioxidant properties [We found a linear correlation between antioxidant activity and the levels of anthocyanins, flavonols, chlorogenic acid, proanthocyanidins, triterpene compounds, and phytosterols. The total amount of proanthocyanidins had a strong positive correlation with the antiradical activity determined by the ABTS method and the reducing activity determined by the FRAP method . We found that the total triterpenoid content had a moderate positive correlation with the antiradical activity determined by the ABTS method and the reducing activity estimated by the FRAP method . A moderate correlation was found between the total phytosterol content in cranberry fruit extracts and the in vitro antiradical and reducing activity of their extracts . We found no correlation between the antioxidant activity of the extracts and the anthocyanin or chlorogenic acid content.The total amount of flavonols had a moderate positive correlation with the antiradical activity determined by the ABTS method and the reducing activity estimated by the FRAP method  and flavonols  [p < 0.05) and the FRAP  methods [p < 0.001), and by the ABTS method was r = 0.635 (p < 0.001) [Oszmiański et al. investigated the linear correlation between antioxidant activity and the levels of triterpenoids, anthocyanins, flavonols, and phenolic acids found in cranberry samples of the cultivars ‘Ben Lear’, ‘Franklin’, ‘Howes’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ . The aut< 0.001) .Cranberry fruit is a valuable plant material accumulating flavonoids and triterpenoids. In plant materials, it is often not possible to attribute the biological effects to a specific group of compounds or to a specific compound, as the complex of polyphenolic compounds acts synergistically ,64. As tVaccinium macrocarpon Aiton) fruit samples of different cultivars grown in Lithuanian climatic conditions, in the collection of VU Botanical Garden. The variables of meteorological data (precipitation (mm), temperature (°C), and sunshine duration (h)) in the Vilnius region during 2021 are presented in The object of the study was large cranberry . Cranberry fruits were collected during berry ripening six times: on 12 August, 27 August, 10 September, 24 September, 10 October, and 22 October 2021 . The fruit shapes and sizes of the cranberry cultivars ‘Baifay’, ‘Early Black’, ‘Howes’, ‘Pilgrim’, ‘Red Star’, and ‘Stevens’ are shown in The harvested cranberries were crushed and frozen at −20 °C in a freezer. Afterwards, the samples were transferred to a −60 °C freezer . Cranberry samples were freeze-dried by Gudžinskaitė et al. described methodology . Freeze-v/v) ethanol containing 1% hydrochloric acid in ratio 1:20 according to Urbstaite et al. described methodology [Ethanol extracts were prepared for the determination of the content of flavonols, anthocyainis, chlorogenic acid and proanthocyanidins. The freeze-dried powders of each cranberry samples were extracted three times with 70% . The analysis of the composition of anthocyanins, phenolic compounds, triterpenic compounds, and phytosterols in cranberry fruit was performed using a Waters ACQUITY Ultra High-Performance LC system  with a photodiode array detector. The composition of anthocyanins and anthocyanidins in cranberry samples were determined using methodology described by Vilkickyte G. et al. . Determi2 = 0.9994) and expressed as mg/g (–)-epicatechin equivalent (EE) dry weight.The content of proanthocyanidins was performed using the DMCA assay methodology described by the Heil et al.  and modi∙+  antioxidant activity determination was performed using the methodology introduced by Re et al. [2 = 0.999) which was prepared using standard Trolox solutions of 2000–11,000 μmol/L concentration. The ABTS∙+ radical cation decolorization activity of the acetone cranberry extract was calculated from the standard curve  which was prepared using standard Trolox solutions (375–3000 μmol/L).An ABTSe et al.  and desce et al. . A volum2 = 0.995) which was prepared from standard Trolox solutions . The reducing activity of the acetone cranberry extract was calculated from the standard curve  which was prepared from standard Trolox solutions of 375–2000 μmol/L concentration. The FRAP (ferric reducing antioxidant power) assay was performed by Raudone et al. described methodology . A volumv/v) , formic acid , acetonitrile , hydrochloric acid , and 4-(Dimethylamino)cinnamaldehyde . Reference standards for maslinic acid, corosolic acid, oleanolic acid, ursolic acid, β-Amyrin, campesterol, α-Amyrin, β-Sitosterol, chlorogenic acid, myricetin, quercetin-3-rhamnoside, quercetin-3-α-L-arabinofuranoside, and quercetin-3-α-L-arabinopyranoside were obtained from Sigma-Aldrich . Reference standards for delphinidin-3-galactoside, cyaniding-3-galactoside, cyaniding-3-glucoside, cyaniding-3-arabinoside, peonidin-3-galactoside, peonidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-arabinoside, cyanidin chloride, peonidin chloride, malvidin chloride, and myricetin-3-galactoside were purchased from Extrasynthese . Reference standards for quercetin-3-galactoside and quercetin were obtained from Carl Roth . The reference standard for quercetin-3-O-glucoside was obtained from Biochemistry . In addition, the following reagents were used: Trolox , ABTS 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), sodium acetate , potassium peroxydisulfate, TPTZ -s-triazine) , and ferric (III) chloride hexahydrate . The purified deionized water used in the experiments was prepared using a Milli-Q®  water purification system.The following reagents were used in the study: acetone, methanol , ethanol 96%  and SPSS Statistics 21 . During the study, means and standard deviations (SD) of the three independent evaluations were calculated. To evaluate the difference in the amounts of anthocyanins, anthocyanidins, flavonols, chlorogenic acid, proanthocyanidins, and triterpenic compounds between fruit samples of different cranberry cultivars, the one-way analysis of variance ANOVA with Tukey’s test for multiple comparisons was used. Differences at relation .The fruit samples of the large cranberry cultivars grown in the collection of Vilnius University Botanical Garden were investigated and the phytochemical composition of anthocyanins, flavonols, chlorogenic acid, proanthocyanidins, triterpenoids, and phytosterols during the ripening period of the fruit was evaluated. The timing of cranberry collection is one of the relevant factors determining the biologically active compounds quantity in the cranberry raw material. Furthermore, the different characteristics of cranberry cultivars also have a significant effect on the cranberries’ bioactive compounds composition. Cranberries of ‘Early Black’ and ‘Howes’ cultivars at different maturity stages contained high amounts of anthocyanins, proanthocyanidins, flavonols and triterpenoids, as well as the fruit extracts of these cultivars demonstrated high levels of antioxidant activity. Cranberry raw material prepared at the beginning of berry ripeness (in August) have higher levels of flavonols and proanthocyanidins, while cranberry raw material harvested at the end of the ripening period (in October) have high levels of anthocyanins and anthocyanidins. The data of the study provide new information on the phytochemical composition of biologically active compounds and patterns of their biosynthesis and accumulation in fruit samples of cranberry cultivars grown in Lithuania and allow for determining the qualitative and quantitative composition of biologically active compounds. These results can be used to provide recommendations for further introduction, selection, and cultivation of cranberries in the Lithuanian climatic conditions. Moreover, studies on the phytochemical composition of cranberry cultivars are relevant because they can lead to encourage further research into the effects of cranberry biologically active compounds for health benefits and the use of cranberry plant material in the development of new drugs.


Bacillus subtilis K1 was 4,060,823 bp in genome size, containing 4,213 genes, with 3,665, 3,656, 2,755, 3,240, 1,261, 3,336 and 4,003 genes annotated in the non-redundant protein sequence database (NR), Pfam, clusters of orthologous groups of proteins (COGs), Genome Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Annotation databases, respectively. In addition, a large number of lignocellulose degradation-related genes were annotated in the genome. The cellulose activity of B. subtilis K1 was higher, exhibiting the highest activity of endo-β-glucanase (24.69 U/ml), exo-β-glucanase (1.72 U/ml) and β-glucosaccharase (1.14 U/ml). It was found that through adding cold-adapted cellulose-degrading bacteriaK1 in the corn straw composting under 6°C (ambient temperature), the average temperature of straw composting was 58.7°C, and higher 86.7% as compared to control. The HA/FA was higher 94.02% than the control and the lignocellulose degradation rate was lower 18.01–41.39% than the control. The results provide a theoretical basis for clarifying the degradation potential of cold-adapted cellulose-degrading bacteria and improving the cellulose degradation efficiency.Degradation of crop straw in natural environment has been a bottleneck. There has been a recent increase in the exploration of cold-adapted microorganisms as they can solve the problem of corn straw degradation under low temperatures and offer new alternatives for the sustainable development of agriculture. The study was conducted in low-temperature (10°C) and high-efficiency cellulose-degrading bacteria were screened using carboxymethyl cellulose (CMC) selection medium and subjected to genome sequencing by the third-generation Pacbio Sequl and the second-generation Illumina Novaseq platform, and their cellulase activity was detected by 3,5-dinitrosalicylic acid (DNS) method. The results showed that the low-temperature (10°C) and high-efficiency cellulose-degrading bacterium  Northeastern part of China is the major grain producing region in China, contributing to 25% grain production and 33% grain transfer amount, which serves as a “stabilizer” and “ballast” ensuring the national food security . The proCrop straws degrade slowly under natural conditions in Northeastern part of China due to the limitation of the overall economic conditions and regional climate characteristics . At presTrichoderma  is required for the degradation of straw cellulose polymers, and these enzyme families degrade carbohydrates through multiple complementation and coordination of oxidative, hydrolytic, and non-hydrolytic activities . Three echoderma , Aspergiergillus  and Penirganisms . In contrganisms . Currentvolution . In addiellulose . Among ttability .via winter corn straw composting experiment, which could provide a theoretical basis for comprehensive utilization of straws.In this study, the bacteria with strong cellulose-degrading ability were screened and isolated using the Congo red carboxymethyl cellulose (CMC) medium, and the genetic basis of their cellulose-degrading ability was revealed through whole genome sequencing and analysis, and their ability of cellulose degradation was measured by means of cellulose activity assay. Besides, new cold-adapted cellulose-degrading strains were prepared based on low-temperature cellulose-degrading strains obtained by previous enrichment culture, and their straw-degrading effect was further validated The appropriate amount of winter forest soil samples from northeast China were collected in natural environments below 10°C, loaded in sterile self-sealing bags, brought back to the laboratory, and refrigerated at 4°C. 10.0 g of the above samples were placed in a triangular bottle with 90 ml of sterile water for 20 min, followed by fully absorbing 1 ml of supernatant into 100 ml of enrichment medium at 10°C, observation, and shaking once a day. When the straw section became loose and soft and the culture medium became darker, it was transferred to the fresh enrichment medium with 5% inoculum and then repeated four times.2PO4 1 g/l,NaCl 0.1 g/l,NaNO3 2.5 g/l, MgSO4·7H2O 0.3 g/l, FeCl3·6H2O 0.01 g/l, CaCl2 ·6H2O 0.01 g/l, and corn straw 5 g/l, pH = 7.0 and the Luria-Bertani (LB) liquid and solid media containing: tryptone 10 g/l, yeast extract 5 g/l, and NaCl 10 g/l, pH = 7.0; Carboxymethylcellulose sodium culture (CMC) medium: sodium CMC 5 g/l, NaNO3 3 g/l, KH2PO4 1 g/l, MgSO4·7H2O 0.5 g/l, KCl 0.5 g/l, FeSO4 0.01 g/l, and agar 15 g/l, pH = 7.0 were prepared and used in this study. The activities of different cellulases in the bacterial solution were detected by 3,5-dinitrosalicylic acid (DNS) method (1), and salicin was the substrate for BG (Hutchinson enrichment liquid medium containing: KH) method . Briefly) method , absorbee for BG .−4, 10−5, and 10−6 were coated on the above isolation medium, and cultured at 10°C for 3 d to obtain pure culture. Then the purified strains were inoculated into the cellulose Congo red medium and cultured at 10°C for 4 d, followed by preliminary screening based on the ratio of the diameter of the hydrolysis circle (D) to the diameter of the colony . Finally, 1 ml of the suspension (1 × 109 bacteria) of the cellulose-degrading strains screened was inoculated into the liquid fermentation medium  and cultured by constant temperature shaking at 10°C and 160 r/min for 6 d, and its enzyme activity was measured  amplification was conducted with 16S ribosomal ribonucleic acid (rRNA) V3-V4 as the amplification region, and 27F (5’-AGAGTTTGATCMTGGCTCAG-3′) and 1492R (5’-TACGGYTACCTTGTACGACTT-3′) were used as forward and reverse primers. Amplicons were examined by 1% agarose gel electrophoresis . A phylo® DNA Isolation Kit. The purity and concentration of the total DNA were measured by virtue of a NanoDrop ND-1000 nucleic acid quantitation instrument. The purified DNA was sent to Wuhan OneMore Technology Co., Ltd. for whole genome sequencing. The genome was filtered and assembled by HGAP 2.0  remove reads with less than a certain percentage of low-quality bases , a certain percentage of reads with N bases (default is 10 bp), overlap exceeds a certain threshold (default is 15 bp) and the possibility reads originating from the host . Coding Bacillus subtilis K1 and the pre-screened composite strains were selected for subsequent straw composting experiment. Corn straws were the composting materials, containing 11.9% water, 0.62% total N, 0.09% total P, 0.28% total K, C/N 37.42, and urea (46% N). Three treatments were set up in the composting experiment:7 CFU/l) of composite strain  and 10 kg of urea. (2) Bio-composting (MC): About 1,000 kg of corn straws were composted, with 2.5 l each of Bacillus subtilis K1 and composite strain fermentation broth and 10 kg of urea. (3) Control (CK): About 1,000 kg of corn straws were composted without adding microbial agents but added with 10 kg of urea. The experiment was repeated 3 times.Bio-composting (M): About 1,000 kg of corn straws were composted and added with 5 l .2O5, K2O and active organic matters and the pH of straws were measured in accordance with China’s organic fertilizer standard (NY 525–2012). The humus components were determined according to Liu Xiaojing’s method Sodium pyrophosphate-sodium hydroxide extraction, potassium dichromate oxidation capacity method  were expressed as SD (standard deviations). Following an Analysis of variance (ANOVA), the means were determined using Tukey’s Honestly Significant Difference (HSD) test at p ≤ 0.05. The data were statistically analyzed using IBM SPSS Statistics 22.0. Origin’s graphics and analysis software was used to create the graphical representations .Every experiment was conducted using a completely random design. The results of three replicates  and entered a high temperature period, reaching the peaks of 68.4°C and 74.8°C, respectively. Moreover, the temperature maintained at above 50°C for 16 d. In MC, the temperature rose rapidly, exceeded 50°C at 3 d, reached the peak (above 70°C) at 10 d and then started to decline, the high temperature above 60°C lasted for 31 d, and then the temperature dropped below 30°C. In M, it took a longer time to enter a high temperature period since long-term normal temperature microbial fermentation agents were used. In CK, the temperature rise during accumulation was slower than that in MC (significantly lower than the highest temperature in MC), and no obvious high temperature period was found, with greater temperature fluctuations. According to the statistical analysis of the grouped data, the temperature had a highly significant difference between MC and M during the whole fermentation process. As observed in p 0.05 according to multiple comparisons  and humic acid (HA) in the combined forms of humus during composting are shown in Bacillus subtilis (NR118383.1), suggesting the highest affinity between K1 and B. subtilis (The 16S rRNA gene amplification (using 27F and 1492R primers) and sequencing were conducted for identification of K1. The 16S rRNA gene sequence of K1 was 1,500 bp in length (GenBank ID: SUB12287217), which was submitted to NCBI for BLAST sequence analysis, and a phylogenetic tree was constructed . The ressubtilis .B. subtilis K1 by the third-generation Pacbio Sequl and the second-generation Illumina Novaseq platform. As shown in B. subtilis K1 was 4,060,823 bp in genome size, containing 4,213 genes, 10 5S rRNA, 10 16S rRNA and 10 23S rRNA, with 3,665, 3,656, 2,755, 3,240, 1,261, 3,336 and 4,003 genes annotated in the NR, Pfam, COGs, GO, KEGG and Annotation databases, respectively. Furthermore, the genome circle graph of B. subtilis K1 was drawn through TBtools software based on the genome-related information , C and COG2723 (BG) were the most abundant COGs. The high diversity of functional annotations suggested that B. subtilis K1 possesses the genetic basis for cellulose degradation.Total 2,775 genes were annotated in COGs, dominated by carbohydrate transport and metabolism (8.82%), amino acid transport and metabolism (9.91%) and transcription (8.07%) . To clarIt was indicated in GO annotations that genes were mainly enriched in cellular components, followed by molecular functions and biological processes. In terms of cellular components, plasma membrane (926 genes), integral component of membrane (726 genes) and cytoplasm (478 genes) were the most important pathways . With reB. subtilis K1 genome, accounting for 2.99% of all coding genes. Among them, glycoside hydrolase (GH) was crucial for the degradation of carbohydrates, and 47 GH genes were annotated in the genome  genes (In 8) genes . The proB. subtilis K1, enrichment of culture by cold domestication was utilized. The cellulose activity of B. subtilis K1 was higher, among which β-glucanase (24.69 U/ml), exo-β-glucanase (1.72 U/ml) and β-glucosaccharase (1.14 U/ml) were found to have the highest degradation activity. B. subtilis K1 is a regular single cell, 2–5.5 μm long and ~ 0.3–0.4 μm wide, with flagellum around its body as shown in its SEM micrograph. In addition, it is a small, round, moist, smooth, viscous, easy to pick, having G+ rod, milk-white round colony with the growing conditions of 5–40°C , pH 3.3–8.5 , optimal carbon source of sodium CMC-Na. It entered the logarithmic period at 8 h, with the highest OD600 value at 30 h, and then entered the stabilization period. Despite a strong cellulose-degrading ability, fungi grow more slowly than bacteria, and their cellulase has poor alkali resistance , the average temperature of straw composting was 58.7°C higher 86.7% than the control. The HA/FA was higher 94.02% than the control and the lignocellulose degradation rate was lower 18.01–41.39% than the control. Correspondingly, different treatment C/N ratio dropped from 37.25 to 21.40%、21.92% throughout the process. Given the low temperature in Northeastern China, the microbial diversity is high in the straw incorporated soil and the rotten straws, and many types of microbes may degrade the main parts of plants and release nutrients into the environment, thereby participating in nutrient cycling  in the cellulose degradation pathway, and these genes were different in the expression, resulting in different expressions of CMCase, C1, and BG in the strain. These three enzymes act synergistically to completely hydrolyze cellulose GHS family cellulases function as endocellulases and/or exocellulases and play a key role in the early stage of cellulose degradation , exo-β-glucanase (1.72 U/ml) and β-glucosaccharase (1.14 U/ml) showed highest activity. It was found through the winter corn straw composting experiment that under an average outside temperature of 6°C, after the cold-adapted corn straw decomposing bacteria were added, the average temperature of straw composting (58.7°C) was 86.7% higher than that of the control. The HA/FA ratio in humus was increased by 94.02%, and the degradation rate of lignocellulose was lower 18.01–41.39% than the control. The study results provide a theoretical basis for clarifying the degradation mechanism of cold-adapted straw-degrading bacteria and improving the cellulose degradation efficiency and have instructive significance for the development of high-efficiency straw-degrading bacteria.The low-temperature (10°C) and high-efficiency straw-degrading bacterium The datasets generated for this study can be found in NCBI public database. All sequences of 16S rRNA genes and whole genome sequence can be found in Sequence Read Archive (SRA) under Bio Sample accession no. PRJNA902081.YJ and ZH planned and designed the research. ZH performed research and contributed to the methodology, writing, and editing. HL, YZ, and XW assisted in analysis, and compiled the data and results. HD and YH helped in experiments and improved the manuscript. QA, BD, and YJ critically revised the manuscript. All authors contributed to the article and approved the submitted version.This work was funded by the LiaoNing Revitalization Talents Program (Project no. XLYC1807221) and Shenyang City Science and Technology Plan (Project no. 21–109–3-01).PD was employed by General Manager, Contec Global Agro Ltd., Nigeria.The remaining 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.


Sleeve dilatation after laparoscopic sleeve gastrectomy (LSG) causes weight regain (WR). Banded sleeve gastrectomy (BSG) was proposed to prevent dilatation and reduce WR.A retrospective cohort study on patients who underwent BSG and LSG and completed 4 years of follow-up from 2016 to 2021 was included. Body mass index (BMI), percentage of excess weight loss (%EWL), percentage of total weight loss (%TWL), and FT scores were calculated at 1, 2, 3, and 4 years. The sleeve volume was estimated at 6 months, 1 year, and 4 years. Multi-variate analysis was conducted to assess correlations between covariates. WR was calculated as weight gain > 10%, > 10 kg above the nadir, or BMI increase of ≥ 5 kg/m 2 above the nadir.p. < 0.001) with no significant main effect of surgery (p.0.438). Mean sleeve volume at 6 months was 102.32 ± 9.88 ± 10.28 ml in LSG vs. 101.89 ± 10.019 ml in BSG and at 4 years was 580.25 ± 112.25 ml in LSG vs. 157.94 ± 12.54 ml in BSG (p. < 0.001).This study included LSG 1279 patients and BSG 132 patients. Mean %EWL at 1 year was 83.87 ± 17.25% in LSG vs. 85.71 ± 7.92% in BSG and was 83.47 ± 18.87% in LSG and 85.54 ± 7.48% in BSG at 4 years. Both had significant weight loss over time (p.0.002) and 31 (2.4%) vs. zero (0%) (p.0.07) using the > 10%, > 10 kg increase above the nadir and the ≥ 5 kg/m 2 BMI increases above the nadir formulas, respectively.WR occurred in 136 (10.6%) and 4 (3.1%) (p.0.002) in LSG and BSG patients, 90 (7%) vs. zero (0%) (BSG had significantly lower sleeve volume, significantly lower WR, and significantly lower FT scores than LSG after 4 years from surgery; however, volume changes were not correlated with weight loss. Laparoscopic sleeve gastrectomy (LSG) has remained the most common bariatric procedure worldwide since 2014, according to the IFSO global registry . Higher Even though the role of sleeve dilatation in WR after LSG is still uncertain, dilatation of the sleeve may be an important cause of WR after LSG as loss of restriction allows more food intake , 7. WR hSome studies proposed banded sleeve gastrectomy (BSG) by applying a band around the upper part of the sleeve pouch to prevent its expansion, hence maintaining better weight loss . Some stRadiological assessment of the sleeve volume is a useful tool to report sleeve dilatation accurately . This voThe study aimed to compare the outcomes between LSG and BSG in terms of sleeve volume, weight loss, WR, and food tolerance (FT) over an intermediate follow-up period of 4 years. To our knowledge, there are no published studies that compared these, in addition to the objective evaluation of FT.This was a retrospective cohort study whereby the files of all consecutive patients who had primary LSG or BSG at Madina Women’s Hospital, an IFSO-accredited center of excellence (European chapter), the Radiology department at Main University Hospital, and Medical Research Institute, Alexandria University, Alexandria, Egypt, between January 2016 and December 2017 and who completed a follow-up period of 4 years were included in this study.The details of both procedures were explained to the patients, and they were left to choose. The patients had to pay for the minimizer ring. Bariatric procedures are partly covered by the university, whereby in the other center, the whole procedure was paid for by the patient itself.Written informed consent was obtained from participants. The study conformed to the principles of the Declaration of Helsinki and was approved by the ethical committee of the Medical Research Institute.Weight loss, sleeve volume, FT score, and WR.All the patients were preoperatively screened and indicated for bariatric surgery according to the International Federation for the Surgery of Obesity and Metabolic Disorders (IFSO) criteria.The analyzed data included demographic characteristics and associated medical problems before surgery, operative time, pre-operative workup, and post-operative follow-up.In all cases, routine laboratory tests and nutritional assessment, upper gastrointestinal endoscopy (UGE), and ultrasound (U/S) abdominal examination were performed. Gastro-esophageal reflux disease (GERD) was evaluated using the Los Angeles classification .2 using the formula: IBW = 25 × (height in m)2 . In the BSG group, 25 patients presented with esophagitis preoperatively, and the postoperative routine endoscopy in 21 of them (84%) showed regression of the reflux. No case of gastric band erosion or slippage was noted in our BSG cohort. Three patients (2.3%) in the BSG group experienced solid dysphagia and reflux symptoms during the follow-up period. Endoscopy revealed constriction at the band site that required a few endoscopic pneumatic balloon dilation sessions with satisfactory results.Three cases of gastric leakage in the LSG group were managed by stent insertion with favorable outcomes. The 30-day-postoperative severe morbidity rates were comparable in the study groups with (p.0.736). (Table GERD grade “A” was recorded in 258 (20.2%) patients in the LSG cohort, while it was recorded in 25 18.9%) patients in the BSG cohort (8.9% patip.0.953).The operative time was almost equal in both BSG and LSG cohorts; 41.63 ± 7.45 and 41.82 ± 7.55, respectively (p.0.665); all patients had a crural repair done during their bariatric procedures  patients and 7 (5.3%) patients in the LSG and BSG cohorts  cases, and the BSG 4 (3.1%) according to the formula > 10% above nadir.2 above the nadir, WR occurred in 31 (2.4%) cases in the LSG and zero (0.0%) in the BSG cohort.When the formula weight gain of > 10 kg above the nadir was used, the LSG had 90 (7.0%) and BSG zero (0.0%), and when the formula BMI increase of ≥ 5 kg/mp.0.002) according to the first and second formulas, while according to the third formula, there was no significant difference (p.0.07).WR was significantly higher in the LSG cohort  in the LSG had conversions to RYGB surgery. The indications for conversion were weight regain in 76 (3.9%), GERD in 64 (5.0%), and GERD with WR in 31 (2.2%) patients.Multi-variate linear regression analysis for %EWL change from baseline has concluded coherent findings with mixed design ANOVA results Table  at 1 andThough EWL% change from baseline increased approximately 2%, 0.45%, 0.558%, and 2.2% among BSG than LSG group at 1, 2, 3, and 4 years after surgery, respectively, this difference was not statistically significant.p.003, p. < 0.001, p. < 0.001, respectively).Patients with preoperative BMI ≥ 40 experienced a significant increase in %EWL compared to those with BMIs < 40 at 2, 3, and 4 years after surgery  in BSG compared to the estimated increase of %EWL by 83.72% (95% CI: 81.51 to 85.92%) in LSG (p.0.284). Patients with preoperative BMI ≥ 40 experienced a significant increase in %EWL by 85.59% (95% CI: 83.68‒87.72%) while those with BMI < 40 experienced increases in %EWL by 80.83% (95% CI: 77.46‒84.21%) (p.0.007)  over an intermediate follow-up period of 4 years.In this study, the mean %EWL at one year was 83.87% ± 17.25 in LSG vs. 85.71% ± 7.92 in BSG, and at four years, it was 83.47% ± 18.87 in LSG and 85.54% ± 7.48 in BSG.p =  < 0.001), but no significant differences between both surgical interventions (p = 0.438).Both operations had significant weight loss over time (Two systematic reviews (SR) on %EWL in BSG and LSG reported that at 1 year, the %EWL ranged from 52.4 to 77.4% for BSG and 41.4‒73.5% for LSG, while at 3 years, it ranged from 66.7 to 91% for BSG and 55.9‒73% for LSG, and at 5 years, it ranged from 70.7 to 68.7% for BSG and 53‒57.8% for LSG . NeverthIn general, WR after bariatric surgery is one of the main motives for modifying the established bariatric procedures and the evolution of new procedures . Data fr2 above the nadir weight. In the outcome, the data distribution changed with the different formulas. From the highest incidence with formula 1 (136 vs. 4 cases in LSG and BSG) to lower incidence in the other formulas 2 and 3 (90 vs. 0 and 31 vs. 0 cases). Despite the different incidences, the LSG had significantly more WR after the operation than the BSG. So this suggests that the BSG procedure has equal weight loss after 4 years but prevents better weight regain after the primary procedure and can be seen as an added value for the long term in maintaining a healthy weight.In this study, WR was calculated using three formulas, (1) > 10%, (2) > 10 kg, and (3) ≥ 5 kg/mInterestingly is that different formulas give new insights into the definition and usage of WR. A study by El Ansari et al. in 2021 did a scoping review for weight regain and insufficient weight loss after bariatric surgery. The study concluded that after checking multiple definitions, the inconsistency, multiplicity, and lack of a standardized definition of WR led to poor reporting and understanding of the clinical significance of the WR . Since tLSG is primarily a restrictive procedure that also involves changes in the gut hormones and microbiomes.Nevertheless, the loss of restriction by sleeve dilatation has been correlated to WR after restrictive procedures such as LSG and adjustable gastric band (LAGB) when the dilated pouch allows the patient to eat more food , 6–8. A Furthermore, in banded RYGB, dilatation of the gastric pouch and the gastro-jejunostomy with the loss of restriction have been attributed to WR . Sleeve This study showed to our knowledge, for the first time, in a large cohort of 911 patients in the LSG, the 3.6-fold increase in the stomach after 4 years (580 ml vs. 157 ml) in LSG and BSG, respectively. However, the multivariate analysis revealed that the changes in the sleeve pouch volume did not correlate to the %EWL after 4 years of follow-up, while having a BMI ≥ 40 was the only predictive factor to have a higher average %EWL. Other studies reported similar results with no correlation between the sleeve pouch volume and weight loss , 26, 27.p. < 0.001) after 4 years (23.81 vs. 21.52).Mean FT scores in this study were showing that FT is significantly better in the LSG  has the worst reported FT among bariatric procedures . MoreoveThe lower FT score in the BSG confirms the restrictive feeling over time from the band. In contrast, in the LSG, the increasing size of the stomach can give a more comfortable feeling, affecting the patient’s behavior and eating habits. Therefore, on the forehand, good preparation is necessary with good nutritional and psychological consulting for the period after BSG since this can affect the behavior and feelings of the patients. So, it is a good balancing act for which benefits outweigh which. Results clearly show the advantages against WR and stomach volume. However, the long-term slight disadvantages of the FT should not be forgotten.In this study, grade “A” GERD was recorded in 258 (20.2%) patients in the LSG cohort, while it was recorded in 25 (18.9%) patients in the BSG cohort.It is still debatable if the LSG is a good option for patients with GERD. While some studies reported de novo GERD after LSG, others reported progressive improvement of GERD symptoms over time with weight loss after LSG .During the preoperative consults, the patients from this study chose to proceed with LSG despite being diagnosed with GERD after having a detailed explanation of the advantages and disadvantages of available bariatric procedures. Many patients and surgeons continue to proceed for LSG even with low-grade GERD for its lower morbidity compared to Roux-en-Y gastric bypass (RYGB) .Hiatal Hernia was diagnosed preoperatively by endoscopy in 80 (6.3%) patients and 7 (5.3%) patients in the LSG and BSG cohorts, respectively. Many authors have recommended exploration of the hiatus with crural repair for wide/stretched hiatus , 32. ManThis study used the MiniMizer ring ® for its easy deployment and adjustment to lengths from 6 to 8 cm. Different materials have been used to band the sleeve in BSG, including Gortex mesh, silicon bands, Alloderm grafts, and silastic ring . SilastiThis study had some limitations as a retrospective study that is inherently inferior to prospective randomized controlled studies with possible undetected confounders or bias that was not corrected or measured. Also, this study did not go in-depth on the incidence of reflux, complications, and associated medical problems post-operatively since this is already discussed in a previous study . FurtherBSG prevents sleeve pouch dilatation and maintains significantly lower sleeve volume than LSG. However, the dilatation could not be correlated to the rate of %EWL. BSG was associated with a significantly lower incidence of WR. Moreover, BSG maintained significantly worse FT scores than LSG.